Monday, March 26, 2018

Microarray Rapid Test Speeds up Detection During a Legionella pneumophila Outbreak

In an outbreak of Legionnaires' disease, finding the exact source as quickly as possible is essential to preventing further infections. To date, a standard analysis takes days. Researchers at the Technical University of Munich have now developed a rapid test that achieves the same result in about 35 minutes.

Legionella are rod-shaped bacteria that can cause life-threatening pneumonia in humans. They multiply in warm water and can be dispersed into the air via cooling towers, evaporative recooling systems and hot water systems.

The most dangerous among the almost 50 species of Legionella is Legionella pneumophila. It is responsible for 80 percent of all infections. When an outbreak occurs, the source of the germs must be identified as soon as possible to prevent further infections.

Similar to a paternity test, the origin of the outbreak is confirmed when the germs in the process water of a technical system exactly match those identified in the patient. However, often numerous systems must be tested in the process, and the requisite cultivation for the test takes around ten days.

Faster detection with antibodies

Meanwhile there is a rapid test for detecting the Legionella pathogen in the clinic. It identifies compounds of Legionella in the urine of patients. "Unfortunately, this quick test serves only as a first indication and is not suitable for screening the water of technical systems," says PD Dr. Michael Seidel, head of the research group at the Chair of Analytical Chemistry and Water Chemistry of the Technical University of Munich.

The team of scientists thus developed a measuring chip in the context of the "LegioTyper" project funded by the German Federal Ministry of Education and Research. This chip not only detects the dangerous pathogen Legionella pneumophila but also identifies which of the approximately 20 subtypes is present.

Fast, inexpensive and versatile

The foil-based measuring chip uses the microarray analysis platform MCR of the Munich company GWK GmbH. Using 20 different antibodies, the system provides a complete analysis within 34 minutes.

"Compared to previous measurements, the new method not only provides a huge speed advantage," says Michael Seidel, "but is also so cheap that we can use the chip in one-time applications."

The system can be deployed for environmental hygiene as well as clinical diagnostics. In combination with a second, DNA-based method, the system can even distinguish between dead and living Legionella pathogens. This allows the success of disinfection measures to be monitored. The project participants will present their system to the public for the first time at the Analytica 2018 trade fair in Munich (Hall 3, Booth 315).


Wunderlich, A.; Torggler, C.; Elsaesser, D.; Lück, C.; Niessner, R.; Seidel, M., Rapid quantification method for Legionella pneumophila in surface water. Analytical and Bioanalytical Chemistry 2016, 408(9), 2203-2213 - DOI: 10.1007/s00216-016-9362-x.

Kober, C.; Niessner, R.; Seidel, M. Quantification of viable and non-viable Legionella spp. by heterogeneous asymmetric recombinase polymerase amplification (haRPA) on a flow-based chemiluminescence microarray. Biosensors and Bioelectronics, 2018, 100, 49-55 (This study was made available online in 08/2017 ahead of peer-review and published February 15, 2018.)

More information:

The LegioTyper project is funded by the German Federal Ministry of Education and Research in the context of the "Civil Security - Protection against biological threats and pandemics" program. The antibodies were provided by the German Reference Laboratory for Legionella at the TU Dresden.

The microarray analysis platform MCR of the Munich company GWK GmbH is also used in a test for antibiotic residues in milk developed at the same chair:

HelixBind Awarded Grant to Accelerate Development of Culture-Free Test for Bloodstream Infections

HelixBind, Inc. announced a collaboration with CARB-X, (Combating Antibiotic Resistant Bacteria Accelerator), a global partnership funding the development of promising new antibacterial products and diagnostics. The company is eligible to receive up to $2 million based on certain milestones supporting the development of a test for bloodstream infections on the company's proprietary RaPID diagnostic platform.

RaPID/BSI targets patients suspected of sepsis, a systemic inflammatory response to a bloodstream infections (BSIs). With approximately 30 million cases and 6 million deaths annually worldwide, sepsis is a global healthcare priority. Early and accurate diagnosis is vital to proper disease management as prognosis deteriorates hourly, but current diagnostic techniques relying on culture may take days for results. RaPID/BSI will provide identification of pathogens and resistance mechanisms within hours and direct from blood.

"We believe RaPID will enable clinicians to place patients on targeted antimicrobial days sooner, leading to better outcomes, reduced time in the hospital, and improved antibiotic stewardship," said Alon Singer, PhD, Founder and CEO of HelixBind.

In announcing the award, Kevin Outterson, Executive Director of CARB-X said: "HelixBind's diagnostic project is an exciting addition to the Powered by CARB-X portfolio, building the number of diagnostics in the portfolio and enhancing its diversity.  The ability to identify infections and drug-resistant bacteria in a timely manner will enable doctors to treat patients more effectively and save lives."

About CARB-X

CARB-X is one of the world's largest public-private partnership devoted to early development antibacterial R&D. Funded by ASPR/BARDA and Wellcome Trust, with in-kind support from NIAID, CARB-X is investing up to $455 million from 2016-2021 to support innovative antibiotics and other therapeutics, vaccines, rapid diagnostics and devices to treat drug-resistant bacterial infections. CARB-X focuses on high priority drug-resistant bacteria, especially Gram-negatives. CARB-X operates through Boston University. Other partners include RTI International, the Broad Institute of Harvard and MIT, MassBio, and the California Life Sciences Institute (CLSI).

About HelixBind, Inc.

HelixBind is a privately held company devoted to improving the diagnosis and management of invasive infections.  Its proprietary RaPID platform provides culture-free characterization of bacterial and fungal pathogens and resistance mechanisms direct from patient specimens, enabling clinicians to quickly diagnosis and monitor infection to ensure prompt and appropriate care.

An Old TB Detection Test Gets a Much-Needed Makeover

A new compound called DMN-Tre has been found to skip the errors encountered with other TB detection tests, and is more affordable, quick and versatile.

Looking for mycobacteria in sputum smears remains a common method for detecting tuberculosis (TB), especially in low resource areas. However, these tests are less sensitive and the results, less reliable. Now, by engineering a new fluorescent dye, scientists from the US may have provided a new lease of life to this antiquated method.

In a typical TB detection test, patients are asked to cough up sputum that is then spread as a thin layer on glass slides. Next, a technician adds dyes that bind to the bacteria, making them appear as coloured spots under a microscope. Two dyes, Ziehl-Neelsen and auramine, are used for this exercise.

The problem with confirming a TB infection this way is that “the dyes stain all cells with a very thick cell surface” irrespective of what kind of cells they were, explained Mireille Kamariza, an author of the new study. Because sputum contains many bacteria apart from the TB pathogen, some of these other microbes could also take up the dye and get counted as mycobacteria.

Another source of error is the destaining procedure. For identifying the microbe, it is essential to first remove the excess colour from the slides. This is achieved by rinsing the slides with a decoloriser and water to remove the unbound dye. The step called washing is usually performed multiple times to remove the background stain such that the marked bacteria become visible. But sometimes, after successive washes, some mycobacteria could also lose the stain.

The sensitivity of microscopy based tests varies between 32% and 94% depending on the skill and experience of the technician performing the test. This doesn’t bode well for disease control: TB is contagious and millions of new cases are identified every year. In 2016, 2.79 million were identified in India alone.

A new compound called DMN-Tre (short for DMN-tagged trehalose) can reduce these errors. The success of a method devised with this compound relies on the use of a sugar called trehalose as part of the dye. Only mycobacteria – and not other cells in the human body – use trehalose as fuel, and they also gorge on the dye. They then appear as fluorescent spots under a microscope, confirming a TB infection.

Part of the problem in detecting pathogens stems from the test’s design. Mycobacteria contain an oily substance in their cell walls called mycolic acid. It makes the pathogen’s outer coating unusually waxy to the extent that it impedes dye molecules from binding to this layer. As a result, mycobacteria don’t get stained.

Now, Kamariza and co. have turned this disadvantage on its head. They have tailored a dye that only fluoresces when it enters the waxy cell wall of the bacterium.

“DMN-Tre is a solvatochromic dye – it can ‘sense’ its environment,” Kamariza told The Wire. This means DMN-Tre behaves differently in different solvents. When in water, the dye will not fluoresce. But the minute it encounters a greasy environment, it starts glowing, according to Kamariza.

The researchers stumbled on the possibility of producing such a dye when they were studying trehalose and its function in mycobacteria. To understand the fate of the fuel within the pathogen, they needed tools to track the sugar. The simplest way to do this was by attaching a fluorescent molecule to a molecule of trehalose.

By tracking the sugar, they found that when trehalose molecules entered the bacteria, they became modified and added to the waxy cell wall.

“When I joined the team, I noticed that if we could modify a sugar such that it fluoresces only when inside cells, we could use it as a diagnostic biomarker for TB,” Kamariza said.

DMN serves this purpose. The thing about DMN is that its structure changes in different solvents. When in an oily environment, it contorts in such a way that it starts glowing. Kamariza and team thought that by attaching DMN to trehalose, they could ensure the DMN came in contact with the mycolic acid in the cell wall. And that when that happened, the DMN would fluoresce.

It did.

Another advantage of working with trehalose is that it is specifically used by TB pathogens and not by other microbes usually present in the human body. While some other disease-causing bacteria also utilise trehalose, they are not present in human sputum and don’t risk affecting test results.

Once DMN-Tre was synthesised, scientists tested its diagnostic efficiency against other fluorescent dyes used to detect TB.

Kamariza and co. first assessed the dye on bacterial strains grown in the lab. While DMN-Tre appeared to be very specific for mycobacteria and could stain more than 70% of them under 30 minutes, the scientists found that those strains that were growing at a faster rate also had the most intense colour under a microscope. This implied that the test could distinguish between live and dead bacteria – an ability beyond the scope of other dye-based methods.

To confirm this, they deliberately killed a group of mycobacteria by heating them, and checked if DMN-Tre still fluoresced by combining with the cell wall. It didn’t.

Similarly, when the scientists added antibiotics to the sputum sample before staining it with DMN-Tre, there was a perceptible change in the intensity of fluorescence. A period of three hours was enough to alter the staining pattern. This has many implications, especially in detecting the drug-resistant variety of TB that makes up 47% of all cases in the world.

Patients carrying multiple-drug-resistant (MDR) TB do not respond to common drugs like rifampicin. Their condition can also remain undiagnosed until the diseases has reached terminal proportions. This plagues both treatment and control strategies.

Sumona Datta, a clinical research fellow at the Imperial College London, said, “It’s not that we don’t have treatment options against the drug-resistant type of TB. But typically, such drugs are more expensive, more toxic, less powerful and take a longer duration for treating the infection.”

As a result, medical practitioners prescribe drugs to treat a drug-resistant infection only after MDR-TB has been confirmed or when patients don’t appear to respond to a combination of two or more drugs. Unfortunately, the required therapy is often administered too late to make a difference to patient health.

“That is why the new study with DMN-Tre is more exciting,” said Datta, who was not involved with the study.

Using DMN-Tre, scientists can now tell if the mycobacteria strain is resistant to certain drugs within a day, at least in theory. This is a promising and inexpensive alternative to other methods of TB detection, some of which also require sophisticated instruments and can’t be used in resource-poor settings.

“The revolutionary thing here is being able to track treatment response,” Carlton Evans, a professor of global health and Datta’s colleague at Imperial College London, told The Wire. “The greatest promise is in studying if patients respond to therapy and when they become non-infectious.”

None of the existing dyes can guide such therapeutic decisions. That the proposed technique also takes under two hours to finish and is as sensitive as auramine makes DMN-Tre particularly useful.

However, some limitations that apply to microscopy-based tests also apply to DMN-Tre testing.

For example, to be able to see the bacteria under a microscope, 1,000-10,000 bacteria are required per millilitre of sputum, according to Datta. “If this condition is not met, then a large proportion of people with true TB may still be categorised as uninfected.” So the tedious process of culturing the bacteria is still the gold standard to achieve an accurate analysis.

Further, “this test cannot detect extrapulmonary TB,” Dutta said, referring to cases where the mycobacteria are present in organs other than lungs. It has been estimated that 10-25% of TB infections worldwide are of this kind. In India, the numbers can be anywhere between 30% and 50%. In infections of this type, the bacteria do not show up in the sputum and could evade detection.

But the most pressing issue is DMN-Tre’s usability in the field. So far, the method has been tested in 16 patient samples, all of which turned up positive for TB. During the analysis, DMN-Tre staining proved to be rapid and efficient for detecting mycobacteria in patient samples without the need for multiple washing.

The results look promising to Datta but she has a few reservations. For their field analysis, the scientists first decontaminated and centrifuged the sputum samples before staining them with DMN-Tre. “Both these steps – decontamination and centrifugation – kill other bacteria more than they affect mycobacteria, and can concentrate the TB pathogen in the sample,” she explained. So this in itself adds to the test’s success by eliminating other contaminating bacteria.

But since centrifugation and decontamination both require sophisticated equipment, the method may not be as successful in settings with basic clinical resources, per Datta.

T. Jacob John, an emeritus professor at Christian Medical College, Vellore, echoed the sentiment. While he agreed that DMN-Tre has a few potential advantages over traditional staining techniques, he said that larger tests would have to be conducted, and their results evaluated against the ease of conducting them in ordinary labs at low costs. “Only if these conditions are fulfilled will this method become popular and be recommended for wider use,” he said.

Akonni Biosystems Announces Strategic Partnership with Major Chinese Diagnostics Company Righton

Akonni Biosystems Inc. and Chinese in vitro diagnostics company, Righton, announce a commercial agreement that enables Righton to sell Akonni's products for nucleic acid purification and molecular diagnostics to researchers, clinical laboratories, and hospitals in China. The products to be sold include TruTip®, Akonni's best-in-class sample preparation technology, which is low-cost and simple, yet highly effective at purifying DNA and RNA from the most difficult sample types. In addition, for diagnostic applications, Righton plans to commercialize Akonni's TruDiagnosis® system based on the proprietary TruArray® multiplexed diagnostic solutions. Righton has made an equity investment of $7.5 million USD in Akonni's series D fundraising round bringing the total investments in the Series D round to $13.9 million USD.

The two-way license agreement also grants Akonni exclusive rights to Righton's extensive molecular diagnostic product portfolio outside of China. Righton has established itself as a major player in the >$1 billion USD molecular diagnostics industry in China, with 28 CFDA-approved diagnostic tests, a well-established customer base, distribution and manufacturing infrastructure, and 2017 annual revenues exceeding 180 million RMB. Righton's established diagnostic tests utilize a combination of real-time polymerase chain reaction (PCR), next-generation sequencing (NGS) and droplet digital PCR (ddPCR) technologies, etc., focusing on applications such as Leukemia, Lymphoma and solid tumors in Oncology, and 30 different infectious diseases. They have an established customer base in more than 400 tertiary hospitals in 29 provinces in China.

"Righton has developed an impressive, well-established menu of reliable, affordable diagnostic tests and an outstanding reputation in China," said Dr. Charles Daitch, CEO of Akonni Biosystems. "We see a lot of value in their products that we believe can offer significant improvements to healthcare communities, and we are very excited for the opportunity to expand access to these products globally." Daitch pointed out that the companies share great synergies in their mission, vision, company culture, products and technologies, which both companies believe will enable them to expand their customer reach and accelerate revenue growth. The companies plan to initiate joint development projects for applications such as liquid biopsy testing in Oncology, non-invasive prenatal testing (NIPT), companion diagnostics and multiplexed panel testing for infectious diseases. "The investment from Righton provides us the financing we need to further our commercial goals outside of China, including the submission of our first US FDA application for our TruDx® 2000 platform. In addition, the partnership will lead to significant commercial revenues from China for both Akonni and Righton in the near-term," Daitch said.

Dr. Xiong Hui, founder and CEO of Righton, said, "While we have developed an extensive product portfolio and very successful business, we see a great opportunity to partner with Akonni. The attributes and flexibility of Akonni's platform, combined with its low cost, should allow us to break into a very large segment of the China market." Righton sees particular value in being able to use and adapt Akonni's products in China, highlighting that the two companies will work together on design and development, ensuring that the products meet the particular needs of Chinese customers and patients.  

Akonni believes the strategic partnership will help it achieve further cost savings in raw materials, manufacturing and production costs. "Whereas many small diagnostic companies choose to sell direct, we believe that our strongest path forward to commercialize Akonni's products and provide the most value to our customers is through a strategic partnership with an established company in China," said Michael Reinemann, Director of Business Development for Akonni. "Righton knows the customers, and the intricacies of the Chinese market segments, local geographies and regulatory and reimbursement environments, and can help us achieve a much more favorable cost of goods in a timely manner."

In addition to supporting Righton's core Oncology business, Righton will support the China launch of Akonni's highly anticipated Tuberculosis (TB) diagnostic product lines. According to the World Health Organization (WHO), despite significant improvements in TB control efforts recently, China still represents one of the largest TB burdens in the world. China has an estimated 1 million new cases of tuberculosis each year – more than any other country except India – including an estimated 63,000 cases of the highly dangerous, costly and deadly multidrug-resistant Tuberculosis (MDR-TB). China currently accounts for nearly one third of the MDR-TB globally. Akonni has invested heavily in its TB and MDR-TB diagnostic tests and is a major player in this market through grants from the Centers for Disease Control and Prevention (CDC) and National Institutes of Health (NIH), and collaborations with TB experts at Harvard University, the University of California San Diego (UCSD), and Johns Hopkins University.  

Scientists Develop Rapid Test that Determines Antibiotic Resistance

Scientists in Germany have developed a test using electric fields and Raman spectroscopy to ascertain specific antibiotic resistance of pathogens and thus enable the prescription of effective treatment.

Scientists of the Leibniz-Institute of Photonic Technologies (Leibniz-IPHT), Center for Sepsis Control and Care at the University Hospital Jena and Friedrich Schiller University are working on a faster and cheaper alternative for hitherto time-consuming pathogen diagnostics. Commenting on the benefits of the new test, Project Manager Prof. Ute Neugebauer said: “We combine light-based analytical methods with microfluidic sample processing.

With our Lab-on-a-Chip system – thus a miniaturised lab – we are able to clearly identify bacterial strains and their resistances, in less than three hours”.

Standard practices for infectious diagnostics require up to 72 hours to ensure a reliable result, owing to the small number of pathogens in a patient’s sample being below the level required to conduct tests. Analysis is therefore only possible after time-consuming cultivation; which is especially restrictive in clinical application during treatments for severe infections such as sepsis, where time is a crucial factor.

Currently, intensive physicians are confronted with an alarming dilemma between making every effort to save a patient and perpetuating the problem of antibiotic resistance. Commenting on the situation, Prof. Michael Bauer, director of the Clinic of Anesthesiology and Intensive Care at the University Hospital Jena, said:

“Far too often we have to administer broad-spectrum antibiotics ‘blindly’, because we can neither analyse pathogen nor potential resistances. Therefore, we possibly use a sledge-hammer to crack a nut. A vicious cycle that aides the development of new resistances” .

Test facilitates a speedy targeted approach

The new method from Jena provides much faster diagnosis that provides a basis for a decision of a reliable therapy. Ute Neugebauer, who works at Leibniz-IPHT and the University Hospital Jena points to tiny electrodes that are fixed on the surface of a stamp-sized chip: “Electric fields secure bacteria in a very small area”. Jena’s scientists then apply various antibiotics in different concentrations on the trapped bacteria and examine them by means of Raman spectroscopy. “This means that we irradiate the pathogens with laser light and evaluate the scattered light spectrum”, says Neugebauer.

Prof. Jürgen Popp, director of the Leibniz-IPHT and head of the Institute of Physical Chemistry of the Friedrich-Schiller University Jena, explains further: “After two hours we can already detect distinct changes in the Raman spectra. Out of these, we can derive whether the strain is resistant or sensible. At the same time, we get information on the required concentration of the antibiotic to constrain bacterial growth. This is an important diagnostic parameter that influences the success of a treatment decidedly”.

The combination of fast, light-based diagnostics and a high automation level reduces the time from sampling to result from currently 72 to just three and a half hours. “Such a fast procedure could revolutionise diagnostics of infectious diseases”, says Prof. Bettina Löffler, director of the Institute of Medical Microbiology at the University Hospital Jena. Currently, researchers are working to develop a platform for application of the test in hospitals. Another, more far reaching, aim is the further development into a cartridge-based rapid test system, to enable general practitioners to identify resistances in a fast and easy way for the first time. This will arm physicians with a powerful tool to aid their endeavours in personalised therapy and the administration of a fitting drug.

The results of the team of chemists, physicians, and biologists were published in the current edition of Analytical Chemistry.

FDA Breakthrough Device Designation for Blood Infection Test

The FDA’s drive to accelerate access to new medical device technologies continues, with a new product from 3i Diagnostics receiving a Breakthrough Device Designation.

While the US regulator first launched the similar Breakthrough Therapy Designation for innovative medicines six years ago, the similar fast-track for devices was formalised in late 2017.

There has been a flurry of activity from the FDA since the start of 2018, with a new AI diagnostic for diabetic retinopathy and a biodegradable patch for surgical wounds already added to the programme.

3i’s technology is called Biospectrix, and detects and identifies bacteria directly from whole blood in less than an hour – a much faster time than standard lab tests, and a technology which could help doctors intervene and provide the right treatment before the infection worsens.

Biospectrix does not require culturing and identifies a broad range of bacteria in less than 60 mins directly from the patient sample, as opposed to the 2-6 days currently needed for culture.

Faster use of an appropriate antibiotic could help cut bacteria-related deaths complications, and treatment costs.

3i Diagnostics was set up by Rajesh Krishnamurthy in February 2013, and he is now its chief technology officer (CT0). Trained in chemical engineering, he previously held senior positions at drug developers Zyngenia and ImmunoGen, and spent seven years in a leadership role at Human Genome Sciences.

“We view this diagnostic as becoming a vital part of a toolkit that can be used globally to aid physicians in making treatment-related decisions i.e. if an antibiotic is necessary and, if yes, the appropriate antibiotic to use.” said Rajesh Krishnamurthy, CTO of 3i Diagnostics, Inc.

“We’re pleased to have been granted Breakthrough Device Designation for Biospectrix and look forward to working closely with the FDA to bring this diagnostic to the market as quickly as possible to address the huge unmet need for rapid broad-range bacterial identification without culturing” said James Janicki, CEO of 3i Diagnostics.

“We see Biospectrix as being a key component of the global strategy to combat the antimicrobial crisis.”

Another benefit of the technology could be in detecting sepsis, a potentially deadly complication of an infection, often bacterial, which could be prevented by faster tests.

Data from the NHS in the UK shows that there are 44,000 deaths every year from sepsis in the UK, with 14,000 of these preventable.

37% of patients that need antibiotics for sepsis are not getting them within an hour, and recent investigations found many hospitals are not carrying out screening on all patients.

3i Diagnostics’ device is also a potential candidate for England’s new Accelerated Access pathway for “strategically important, transformative products” which will be launched in April.

U of M-Developed Tool for Rapid Infection Diagnosis Nears Commercialization

A University of Minnesota chemistry professor who has developed a method for rapidly detecting the presence of urinary tract infections said she’s launching a commercialization push for a new consumer product.

Associate Professor Valerie Pierre of the U’s College of Science and Engineering is a molecular chemist whose main work is developing new kinds of florescent metal-based “contrast agents” for imaging technologies such as PET (positron emission tomography) scanners. These new compounds, when administered to patients before the procedure, could potentially enable scanners to quickly and accurately diagnose many kinds of bacterial infections in vivo, without the need for performing time-consuming cultures.

If that work is successful, it could be a major game-changer since it now takes 48 to 72 hours for medical labs to clinically diagnose any kind of infection from bacterial cultures — including fast-moving and potentially lethal infections. The delay means doctors must now make decisions based solely on symptoms rather than actual diagnoses, inevitably leading to many unnecessary and costly prescriptions for strong antibiotics.

While Pierre and her lab colleagues at the U of M have been working on improving the diagnostic capabilities of PET scanners in recent years, their efforts have also led them onto a related entrepreneurial path for a more modest but still much-needed application – a cheap, easy-to-use diagnostic tool to rapidly diagnose urinary tract infections.

Their new product is a “dipstick”-like, urine-sampling device that could one day be available to consumers in drugstores, much like home pregnancy tests. As with the scanner technology, it uses a fluorescent, metal-based complex which “lights up” when binding to bacterial targets, but in this case is refined to detect the presence of E. coli – the cause of the vast majority of urinary tract infections – as well other kinds of bacteria, all within a matter of minutes.

The inventors said the test would also be able to determine if the bacteria found are resistant to antibiotics, and so suggest the proper kind of treatment.

Pierre told TCB there’s a major unmet need for rapid analysis of all kinds of bacterial infections, but especially so for those of the urinary tract.

“Urinary tract infections are the most common infections in the world, and they especially affect women, so you’re talking about millions of cases each year just in the United States,” she said. “It also happens to be a type of infection which is not very well-diagnosed.

“Most of the time, you call a nurse or a doctor on the phone and they will prescribe antibiotics based on the symptoms, not on actual clinical diagnostics. The reason for that is there’s really nothing out there that works that is accurate and is quick. Our goal was to do something that was just as accurate as a bacterial culture but instead of taking 48 hours, it would take five minutes.”

Her path to commercializing the device was helped along by the U of M’s Clinical and Translational Science Institute, a unit of its Academic Health Center supported through the federal National Institutes of Health and its Clinical and Translational Science Award program, which seeks to move treatments out of the lab and into the market more quickly. The institute funded Pierre’s early work on the device, advised her on how to assess the medical marketplace and supported her development of a working prototype.

Now she says the effort has reached the point where a commercialization push is imminent.

“It’s something we’re very interested in doing either in partnership with a major medical company or through establishing our own startup company,” she said. “This is something the University would very much like to push forward.”

The path they ultimately take could depend on if a CEO can be found who would be interested in leading a startup firm with a possible ground-breaking product in the diagnostics field.

“I know my limitations,” Pierre laughingly added. “I know I don’t want to be a CEO myself. I’ll stick to the science.”

Friday, March 16, 2018

Faster Meningitis Test Developed in Belfast Could Save Lives

A rapid meningitis diagnostic test developed by scientists in Belfast could be rolled out in hospitals across Britain and Ireland once its two-year research study finishes in 2019.

Clinical scientist Dr James McKenna from the Belfast Care and Social Trust was the lead researcher in developing the new test which can be completed in under 50 minutes, compared with standard laboratory tests, which can take up to 48 hours.

The time saving would reduce the unnecessary treatment of patients with antibiotics while they await results. Meanwhile, those patients who have contracted meningococcal disease will test positive in a much shorter time and can be treated immediately.

Dr McKenna developed the LAMP (Loop Mediated Isothermal Amplification) test, which can quickly and accurately be used by doctors and nurses outside a laboratory environment to detect tiny quantities of bacterial DNA.

Diagnosis of meningococcal disease – which can lead to meningitis and blood poisoning – is particularly difficult for GPs and emergency departments to detect in its early stages.

Condition deteriorates

Standard laboratory test results can take up to 48 hours, so patients can be given antibiotics when they do not really need them and some children could be wrongly sent home from their doctor or a hospital only to come back a few hours later as their condition deteriorates.

Therefore a test that can rapidly diagnose patients in a hospital emergency department or a doctor’s surgery could be extremely beneficial and will potentially save lives, researchers have said.
In September 2017, Dr Tom Waterfield, as part of a Queen’s University and Paediatric Emergency Research UK and Ireland collaboration, started a two-year study at the Belfast Hospital for Sick Children, to assess the suitability of the LAMP test to diagnose meningococcal disease in a hospital emergency department environment.

Laboratory results

The results from his study, testing more than 100 patients so far, will be compared with standard laboratory results and help to develop new decision rules for clinicians and Dr McKenna hopes it will reduce unnecessarily prescribing antibiotics, and improve patient management.

Once the two-year study is complete it is hoped the LAMP test will be rolled out across paediatric units in Britain and Ireland, Dr McKenna says.

Dr McKenna is happy with how the study is progressing and that the test is now commercially available.

“It’s good to see something you have worked on actually being used,” he said.

“The trust has licensed it to a company in the Republic of Ireland, Hibergene.”

Dr McKenna says vaccination rates in Northern Ireland are high so this will reduce the number of meningococcal cases but it also means new clinicians will have less experience of seeing them, so it becomes even more important that they have good diagnostic tests to help them make a decision.

More generally, he is saddened when people choose not to vaccinate their children against preventable diseases (including meningococcal disease), as is seen in the so-called “anti-vaxxers” movement.

“People worked for years to get rid of all these diseases,” he said.

“Anti-vaxxers are endangering their own children and other peoples children.

“It is very, very sad to see these diseases we thought were gone, could have been eradicated, coming back.”

He believes education, finding better ways to communicate the usefulness of vaccines, is key.

Sekisui Diagnostics Announces Launch of Silaris™ Influenza A&B Test

Sekisui Diagnostics announced the launch of the Silaris™ Influenza A&B Test.  The Silaris™ test was developed by Mesa Biotech and utilizes an affordable, polymerase chain reaction (PCR) testing platform specifically for point-of-care (POC) infectious disease diagnosis.

The United States is currently experiencing the most severe flu season in recent years, according to the CDC, with widespread activity and high influenza-like illness (ILI) across the majority of the country. The Silaris test allows urgent care, physician offices, hospitals and other facilities to implement an affordable, CLIA Waived, molecular testing option to diagnose and treat patients quickly which is key to minimizing the spread of the virus.  The Silaris™ test offers the simplicity, convenience and procedural familiarity of traditional visual read rapid tests, while providing the superior sensitivity and specificity of laboratory-based PCR testing.  The test is indicated for use with nasal swabs which are less invasive than other sample types and allows for a more comfortable experience for the patient.

"We are very excited to announce the launch of the Silaris™ product because it brings a unique diagnostic tool to the marketplace.  Healthcare professionals can feel more confident they are treating patients accurately and quickly which is critical, especially during heavy flu seasons," said Robert Schruender, President & CEO of Sekisui Diagnostics. "This system is highly complementary with the rest of our point-of-care product line and illustrates our commitment to deliver innovative products which improve patient care."

Sekisui Diagnostics entered into a strategic alliance with Mesa Biotech in November 2017 for exclusive product distribution rights in the U.S. and Canada.  This allows Mesa to leverage Sekisui Diagnostics' established sales/distribution channels and strong reputation among healthcare providers and provide immediate access to distribution channel partners.

"The influenza product is the first assay on our PCR testing platform to receive FDA clearance," said Dr. Hong Cai, co-founder and CEO of Mesa Biotech. "This is an outstanding validation of our testing platform and we remain focused on additional product development and look forward to a long term commercial collaboration with Sekisui Diagnostics of our expandable system."

About Sekisui Diagnostics

Sekisui Diagnostics is a global diagnostics company committed to improving patient's lives by providing innovative medical diagnostics to physicians and laboratories through a global commercial network.  Product lines include clinical chemistry and coagulation systems, reagents, rapid test kits, point-of-care systems as well as enzymes and specialty bio-chemicals.

About Mesa Biotech Inc.

Mesa Biotech designs, develops and manufactures next generation molecular diagnostic tests, bringing the superior diagnostic performance of nucleic acid PCR amplification to the point-of-care (POC). Mesa Biotech's Accula™ System consists of a portable, palm-sized dock and a disposable, assay-specific test cassette. This patented system will allow healthcare professionals to access actionable, laboratory-quality results at the POC with greater sensitivity and specificity than current infectious disease rapid immunodiagnostic tests. Mesa Biotech's technology development has been funded to date by a series of grants and private equity investments.

Mason Scientists Develop Nanotechnology-based Urine Test for Early TB Detection

Scientists at George Mason University have developed a nanotechnology that for the first time can measure a sugar molecule in urine that identifies tuberculosis with high sensitivity and specificity, setting the stage for a rapid, highly accurate and far less-invasive urine test of the disease that could potentially prove to be the difference between life and death in many underdeveloped parts of the world.

The international team led by George Mason’s Alessandra Luchini and Lance Liotta report in Science Translational Magazine that a sugar molecule called “LAM,” which comes from the surface of the tuberculosis bacteria, can be measured in the urine of all patients with active tuberculosis regardless of whether they have a simultaneous infection with another pathogen (e.g. HIV). The more severe the disease, the higher the sugar concentration in the urine, said Luchini, an associate professor in Mason’s College of Science.

Current methods of detection – skin tests, blood tests and chest X-rays – are often very expensive and not always available in rural settings in lesser developed parts of the world. Urine is considered an ideal body fluid for a TB test because it can be easily and noninvasively collected.

“We can measure now what could never be measured before,” said Liotta, co-director of Mason’s Center for Applied Proteomics and Molecular Medicine.

The World Health Organization estimates that TB kills 1.8 million people annually, most of whom might have lived if the highly communicable bacterial infection had been diagnosed sooner and treated. In 2015, 10.4 million people worldwide fell ill with TB, with an estimated four million of those cases never reported, diagnosed or treated, according to the WHO.

The mortality rate for untreated TB is 68 percent, but that figure drops to just five percent when diagnosed and treated, according to the WHO. Once thought to be under control, the disease is again on the rise around the globe.

Accurate urine testing had been impossible in the past because TB shed markers exist in very low concentrations and are masked by other high-abundant, normal urinary resident proteins. The Mason scientists, aided by a group of Mason students and scientists from Peru, Italy and Johns Hopkins University, discovered that a special copper dye will bind and capture the TB LAM sugar with incredibly high affinity.

“We showed that our technology could be used to measure several different kinds of markers for TB in the urine and could be configured as a rapid test similar to a pregnancy test,” Luchini said.
Their work, supported by grants from the National Institutes of Health and from the Bill and Melinda Gates Foundation, served as the basis for the subsequent breakthrough by a group of Mason students in developing a cheap and less-invasive “dipstick” TB test. The Mason students will eventually head to Peru to begin testing their device with hundreds of patients for their research study.

The Mason University Biotechnology partner Ceres Nanosciences will be commercializing the technology, with the aim of making the test available worldwide.
“We have everything in place to do our best to get this out and help people in the world,” Luchini said.

Detection On The Go For A Common Viral Pathogen: Norovirus

Human noroviruses are a leading cause of gastroenteritis globally and inflict a considerable public health burden. The viruses are easily transmitted, sometimes through foods, but also person-to-person. The virus causes intense vomiting and diarrhea, which can result in horrible outcomes in certain populations – especially the elderly and children. One of the many challenges in controlling this threat is the ability to rapidly detect the virus in people or the environment so that measures can be taken to stop the spread of the virus.

The traditional technique used for this is not readily deployable in many settings where the virus may be found, like restaurants, nursing homes, and food preparation centers. This traditional technique relies upon amplification of a segment of the virus genome, which makes it capable of detecting a very small amount of virus. However, the technique requires a large, expensive piece of equipment, electricity, a freezer for the chemicals used in the reaction, and takes at least one hour to complete. While the virus is so easily transmitted, it takes a significant amount of time to send samples to a lab and then tested for a result. Thus, having a rapid, portable method capable of detecting the virus is crucial to stopping its spread.

Researchers at North Carolina State University and the University of Massachusetts have developed a Norovirus assay using a technique called Recombinase Polymerase Amplification (RPA), that shows promise for achieving this. RPA amplifies the virus genome using enzymes that operate at body temperature without the need for altering the temperature like the traditional technique. Because the method requires maintenance of a general, low, constant temperature, it requires considerably less electricity – meaning that portable solar-powered batteries could be used for it.

In one report, this technique has been applied to another pathogen using a researcher’s body heat to power the reaction. In addition to being portable, the method takes 20 minutes or less to amplify the viral genome – much less than the hour or greater demand of the traditional method. Another advantage of this method was its ability to tolerate other chemicals present in samples that inhibit the traditional method. Because it was so tolerant, the researchers were capable of detecting norovirus in patients’ stool by directly boiling the stool without additional preparatory steps. This saves a large amount of time and additional equipment needed for the sample preparation traditionally used.

Overall, this new method would enable portable detection of norovirus from a patient sample in under 30 minutes total. Because the method is amplification-based, it is better able to detect a small number of viruses in a sample. This ability is crucial for detecting noroviruses in food and environmental samples, as the number of viruses found in the samples is often low.

Future work will involve increasing the ability of the assay to detect additional strains of norovirus. Additional evaluation of the performance of this method when used for food and environmental samples is also being performed. Further optimization of this method and its applicability to other samples is currently underway, and it has the potential to yield powerful results in the fight against this ubiquitous pathogen.

These findings are described in the article entitled Development of a Recombinase Polymerase Amplification Assay for Detection of Epidemic Human Noroviruses, recently published in the journal Scientific Reports. This work was conducted by Matthew Moore and Lee-Ann Jaykus from North Carolina State University and the University of Massachusetts.

T2 Biosystems Lands $2MM Grant for ‘Superbug’ Test

T2 Biosystems won a $2 million grant to develop a test using its T2Dx MR-based technology to detect strains of drug-resistant bacteria or so-called “superbugs.”

The grant is from Carb-X, a Boston University-based public-private partnership aimed at furthering anti-bacterial R&D that’s jointly funded by the U.S. Health & Human Services Dept.’s Biomedical Advanced Research & Development Authority and the Wellcome Trust. The group has a $455 million war chest to deploy until 2021 to back the development of new antibiotics, therapeutics, vaccines, rapid diagnostics and devices to treat drug-resistant bacterial infections.

Lexington, Mass.-based T2 said it plans to develop new tests for its T2Dx instrument for 20 additional bacterial species and resistance targets, focusing on the Centers for Disease Control & Prevention’s threat list of antibiotic-resistant blood-borne pathogens.

“The collaboration with Carb-X will accelerate the development of diagnostic tests to identify major bacterial species and resistance genes faster than ever,” president & CEO John McDonough said in prepared remarks. “Identifying these infections directly from whole blood with T2MR technology will help patients receive the right therapy faster – improving patient outcomes and reducing healthcare costs. We believe these diagnostic advances will also support the development of new antibiotics by accelerating clinical trials, attacking the problem of resistance on all fronts.”

“The ability to rapidly diagnose and treat drug-resistant bacteria will save lives by enabling doctors to treat patients more quickly and effectively than is possible with today’s diagnostic tools,” added Carb-X executive director Kevin Outterson. “Our collaboration with T2 Biosystems enhances the diversity of our pipeline with exciting technology. These products are not ready for use in patients yet, but if approved, they offer great potential to fight against life-threatening, drug-resistant bacteria.”

Qiagen Launches New Bacterial Detection Kit in Europe

Qiagen has announced the launch of its new Artus T. vaginalis QS-RGQ Kit in Europe after received CE Mark approval for the new offering.

The molecular diagnostic test has been optimised for the qualitative detection of the protozoan parasite T. vaginalis in both female and male patients, using real-time polymerase chain reaction methods to analyse clinician-collected vaginal swabs, endocervical swabs and urine samples.

Configured for use with the QIAsymphony SP/AS and Rotor-Gene Q instruments, the test is suitable for assessing both symptomatic and asymptomatic patients, and can aid the rapid diagnosis of a bacteria responsible for 170 million to 190 million sexually-transmitted infections each year.

Trichomoniasis is associated with adverse pregnancy outcomes, infertility, postoperative infections, cervical neoplasia and an increased probability of HIV transmission. It is widely underdiagnosed due to a lack of routine testing and the low sensitivity of current diagnostic methods.

Michael Nugent, vice-president and head of infection and immune diagnostics at Qiagen, said: "This molecular test offers timely and sensitive detection of T. vaginalis in a wide range of sample types from both male and female patients."

Quidel Receives FDA Clearance for Its Point-of-Care Sofia® Lyme Fluorescent Immunoassay for Use with Sofia® 2 Instrument

Quidel Corporation, a provider of rapid diagnostic testing solutions, cellular-based virology assays and molecular diagnostic systems, announced today that it has received 510(k) clearance from the United States Food and Drug Administration (FDA) to market its Sofia Lyme FIA to be used with the Sofia 2 Fluorescent Immunoassay Analyzer for the rapid differential detection of human IgM and IgG antibodies to Borrelia burgdorferi from serum and plasma specimens from patients suspected of B. burgdorferi infection. The test is intended for use with either the Sofia or Sofia 2 analyzer to aid in the diagnosis of Lyme disease.

Sofia 2 is Quidel’s next-generation version of its best-selling Sofia instrumented system. Sofia 2 utilizes the original Sofia’s fluorescent chemistry design while improving upon the graphical user interface and optics system to provide an accurate, automated and objective result in as few as 3 minutes. Sofia 2 also integrates wireless connectivity and its barcode scanner within a smaller footprint than the legacy Sofia instrument. The next-generation Sofia system also comes connected to Virena®, Quidel’s data management system, which provides aggregated, de-identified testing data in near real-time.

Lyme disease is the most common tickborne disease in North America and Europe1. In the United States, Lyme disease is caused by the bacterium, Borrelia burgdorferi, transmitted through the bite of an infected blacklegged tick. (1, 2)

Patients infected with B. burgdorferi may experience symptoms associated with three stages: early localized disease, early disseminated disease, and late persistent disease1. The most characteristic symptom of early localized disease is the appearance of erythema migrans (EM) on the skin1,3. EM may also be accompanied by flu-like symptoms days or weeks after infection3. In the second stage, early disseminated disease, untreated patients may begin to see neurological and rheumatological manifestations, and less commonly, dermatological, cardiac, or ophthalmological manifestations. These symptoms generally appear weeks to months after infection1. If the disease continues to be left untreated, late persistent disease may also follow months or years later with continued progression of manifestations in the joints, heart, skin, and nervous system. (2, 3)

Early detection and treatment of Lyme disease can help resolve symptoms and prevent progression of the disease1. The primary means of identifying B. burgdorferi infection is detection of the body’s IgM and IgG antibody response by way of immunoassay3. Detection of IgM antibodies to B. burgdorferi is generally most significant in the earlier stages of the disease. Conversely, detection of IgG antibodies has proven to be significant for longer periods, as the antibodies may remain detectable years after infection.

"The Sofia Lyme Assay’s 510(k) clearance for use on the Sofia 2 instrument will allow healthcare workers to generate a faster result, thereby accelerating the diagnosis and potential treatment of Lyme Disease for the patient. This is another example of our ability to provide simple, cost-effective solutions for physician offices and hospitals that previously had to wait several days for send-out Lyme results,” said Douglas Bryant, president and chief executive officer of Quidel Corporation.

"We expect that this new product introduction will increase the utilization of our Sofia 2 platform, and could create incremental instrument placement opportunities in the near to medium term.”
The Sofia Lyme Assay was previously 510(k) cleared for use on the Sofia instrument. This new 510(k) clearance allows the assay to also be run on the Sofia 2 instrument. The Sofia Lyme Assay is the fourth 510(k) cleared Sofia test for use on the Sofia 2 system: the Sofia Influenza A+B Assay, the Sofia RSV Assay, and the Sofia Strep A+ Assay were 510(k) cleared and CLIA waived by the FDA in 2017.

1. Wormser, G. P., Dattwyler, R. J., Shapiro, E. D., Halperin, J. J., Steere, A. C., Klempner, M. S., Nadelman, R. B. (2006). The Clinical Assessment, Treatment, and Prevention of Lyme Disease, Human Granulocytic Anaplasmosis, and Babesiosis: Clinical Practice Guidelines by the Infectious Diseases Society of America. Clinical Infectious Diseases, 43(9), 1089-1134.
2. CDC.
3. Aguero-Rosenfeld, M. E., Wang, G., Schwartz, I., & Wormser, G. P. (2005). Diagnosis of Lyme Borreliosis. Clinical Microbiology Reviews, 18(3), 484-509.

A Single Test is Able to Diagnose 8 Tick-Borne Diseases

Scientists from the Center for Infection and Immunity (CII) at Columbia University's Mailman School of Public Health have announced a new test for tick-borne diseases (TBDs) with increased accuracy in Nature: Scientific Reports. The TBD Serochip is a single test that can differentiate between Lyme disease and 7 other TBDs.

The current procedure to diagnose Lyme disease requires 2 separate tests and the criteria for diagnosis is notorious for inaccurate results. The team of researchers from CII along with scientists from the Centers for Disease Control and Prevention, the National Institute of Allergy and Infectious Diseases, Roche Sequencing Solutions, Farmingdale State College, and Stony Brook University, conducted the investigation to find standard test for Lyme disease and other TBDs including Babesiosis, Anaplasmosis, Ehrlichia, and Powassan virus or Heartland virus.

“Each year, approximately 3 million clinical specimens are tested for TBDs in the United States. Nonetheless, the true incidence of TBDs is likely greatly underestimated, as patients with presumed TBDs are rarely tested for the full range of tick-borne agents, and only a fraction of positive cases are properly reported," said Nischay Mishra, PhD, a co-lead author of the study, in a press release.

According to the investigators, the TBD Serochip can test 170,000 protein fragments for the existence of pathogens connected to 8 different types of TBDs. As new TBDs are discovered, scientists will be able to alter the TBD Serochip to detect the new disease in less than 4 weeks’ time. The current version also tests for a newer virus, Long Island tick rhabdovirus, which was discovered in Amblyomma americanum ticks.

Because ticks frequently carry more than 1 type of pathogen, the TBD Serochip can identify if a person is infected with multiple types of pathogens. The investigators report “finding antibodies to another agent in 26% of blood specimens from patients with TBD.” The test will also provide information for scientists to observe antibody responses and interactions between TBD agents on how the disease progresses, as well as how genetically diverse pathogens affect immune response.

"Diagnosing tick-borne illness is a difficult journey for patients, delaying effecting treatment," said senior author W. Ian Lipkin, MD, director of CII and John Snow Professor of Epidemiology at Columbia, in the press release. "The TBD Serochip promises to make diagnosis far easier, offering a single, accurate test for 8 different TBDs. Early detection of infection enables rapid and appropriate treatment."

Glowing Molecule can Reveal Live Tuberculosis Bacteria

A molecule made in the lab may change the way tuberculosis is diagnosed in the field.

Chemically tweaking a sugar molecule known as trehalose lets it slip inside the bacteria that causes tuberculosis (TB) and glow. The method offers a quick, simple way to detect the pernicious bug, and may help counter TB, a deadly lung infection that's particularly common in developing countries.

Howard Hughes Medical Institute (HHMI) Investigator Carolyn Bertozzi and colleagues report the work February 28, 2018, in the journal Science Translational Medicine.

Despite its devastating toll on health, the bacteria behind TB, Mycobacterium tuberculosis, can be hard to spot. Current tests rely on chemical stains that have been around for decades and can be finicky. Estimates put the sensitivity of these stains anywhere from 32 percent to 94 percent.

Better detection methods are sorely needed to combat TB, which killed more than 1.7 million people worldwide in 2016, says Bertozzi, of Stanford University. "If you can't even get an accurate diagnosis, how do you treat people?"

As a chemist, Bertozzi, along with her colleagues, studies the molecules that make up bacterial cell walls. Early discoveries by her lab revealed that some bacteria use sugar molecules called trehalose as building blocks. Bertozzi found the cell walls of M. tuberculosis particularly compelling. "There's some really interesting biology there." She began working with a team of scientists who held personal stakes in tuberculosis research. After a chance encounter at a meeting at Janelia Research Campus, Bertozzi decided to collaborate with study coauthor Professor Bavesh Kana of the University of Witwatersrand in Johannesburg, South Africa.

The researchers realized that trehalose molecules - those cell wall building blocks - might offer a way to flag living M. tuberculosis cells. But first, the team needed to find a chemical beacon that would make the flag visible. One chemical, called DMN, seemed to fit the bill. DMN can glow under certain wavelengths of light - but only when it is out of water. Because the M. tuberculosis cell wall contains a membrane that's a "thick layer of grease," Bertozzi says, it's the perfect place for DMN to light up.

That insight - that DMN was "off" until a cell tucks it into its membrane - was key, Bertozzi says. "It's such a simple thing, but simple things like that make all the difference between something that can be deployed or not."

After linking trehalose to DMN in the lab, the researchers tested their hybrid molecule, called DMN-Tre, on an M. tuberculosis relative. Just as they had hoped, the bacteria grabbed the molecule and, within minutes, incorporated it into their cell membranes, where it began to glow under a fluorescent microscope.

In tests on sputum samples from 16 people with TB, DMN-Tre picked up M. tuberculosis cells in all of the samples. The new technique performed similarly to the standard - but more complex and time-consuming - labeling method based on the Auramine O stain, a dye that sticks to acids in bacterial cell walls.

Other tests showed that DMN-Tre is selective to Actinobacteria, the bacterial phylum that includes M. tuberculosis. Human cells and other types of bacteria, both of which are plentiful in sputum samples, don't incorporate the molecule, the researchers found.

Unlike existing TB detection methods, DMN-Tre can also distinguish cells that are metabolically active from those that are not. Because the molecule relies on bacteria to actively incorporate it into the membrane, only healthy cells are labeled, whereas cells that are compromised by drug treatment do not label as well. That property may allow clinicians to monitor how well treatments are working in people, and perhaps even test whether certain mixtures of drugs would work against specific strains of M. tuberculosis.

More work remains before the molecule is ready for use in the field, Bertozzi says. But she's optimistic that the new method could prove useful in the global fight against TB.


M. Kamariza et al. "Rapid detection of Mycobacterium tuberculosis in sputum with a solvatochromic trehalose probe." Science Translational Medicine.

MiraVista Aims to Improve Testing for Misdiagnosed Histoplasmosis Infections

Histoplasmosis isn’t top of mind for most people, but if you’re a Hoosier, it’s likely that you’ve had it. MiraVista Diagnostics Medical Director and President Dr. Lawrence Joseph Wheat says about half of people living in Indiana have had histoplasmosis, an infection caused by a fungus that can be found in soil containing large amounts of bird or bat droppings. Building on nearly two decades of infectious disease testing, the Indianapolis-based company is on the cusp of major growth that could make detecting fungal infections faster and more efficient.

The Centers for Disease Control and Prevention (CDC) says people can get histoplasmosis after breathing in the microscopic fungal spores from the air. Although most who breathe in the spores don’t get sick, Wheat says the infection can be dangerous for people with weakened immune systems. It’s most common in the Midwest, and Wheat says it’s likely about 90 percent of people living in Tennessee have had the infection.

“[Diagnostics] are really important in areas where these fungal infections are seen,” says Wheat. “If these tests aren’t available, there are [immunocompromised] patients who would die without treatment, or would be much sicker by the time they’re diagnosed and have a more difficult response to treatment.”

Wheat says MiraVista performs the bulk of fungal infection testing for the U.S. Humans comprise about two-thirds of its testing, but the company also tests veterinary patients. Most samples arrive at the Indianapolis facility from reference labs and academic medical centers throughout the country.

Wheat developed the proprietary test in the late 1980s as an infectious disease specialist at the Indiana University School of Medicine. He left the university in 2002 to start MiraVista, which began with just five employees, but has now grown to 50.

The company is anticipating to expand even more quickly as it develops a new kind of test for histoplasmosis. Unlike the enzyme assay that requires high-tech equipment in MiraVista’s clinical lab, Wheat says the new diagnostic “is a rapid test, like a pregnancy test that can be performed in the local lab on small numbers of samples.”

A trial testing MiraVista’s rapid diagnostic is underway in Latin America, with a second scheduled to start soon. Wheat says the standard method in Latin America to detect histoplasmosis is to examine a tissue sample under a microscope, which can be a barrier for easy diagnosis.

“There’s probably more histoplasmosis in Latin America than the U.S.,” says Wheat. “It’s common in most Latin American countries, so a rapid test will be very important there. We’re going to put a lot of emphasis on [the trials] in the next two years to try to make [the rapid test] available there.”

While most of its work focuses on histoplasmosis, MiraVista also tests for blastomycosis and coccidioidomycosis, more commonly known as Valley fever, which can be severe even in healthy people. The National Institutes of Health awarded a grant to MiraVista to also develop a rapid test for Valley fever.

The CDC says about 10,000 cases of Valley fever are reported each year, mostly in the Southwest, but tens of thousands more likely occur and are misdiagnosed, because many patients are not tested for the condition.

Wheat says the company’s growth over the last two years has enabled it to work on demanding projects, such as expanding to Latin America. He expects the number of employees at MiraVista to double—reaching 100—in the next two years as it implements new tests and further develops diagnostics in its pipeline.

“We are doubling the size of our building and recruiting more scientists who can bring new ideas to research. We have facilities here to manufacture kits, develop kits and add new services,” says Wheat. “Our veterinarian practice is probably only a quarter of what it could be, once we implement better marketing. We have a lot going on this year we think will take us to another level.”

For Flu Detection, Just Add Water

As one of the worst flu seasons in years continues to take its toll, new technology developed by a University of Florida research team comes at an especially timely moment: It’s the first air sampler that can consistently capture the influenza virus, making it up to 100 times more effective than existing samplers.

That early-detection capability can give health officials the extra time they need to put safety precautions in place, potentially spelling the difference between a small outbreak and a pandemic.

It could also thwart a biological terrorist attack, said Chang-Yu Wu, a professor and head of the environmental engineering department in UF’s Herbert Wertheim College of Engineering.

Wu, who heads UF’s Aerosol and Particulate Research Laboratory, said he became aware of the shortcomings of existing air samplers, typically deployed in large public places such as airports and state parks, about 10 years ago. The problem, he said, was that most samplers were designed to capture relatively large bacteria and other airborne particulate matter pollutants.

“Viruses are considerably smaller than bacteria, so those samplers are only about 10 percent effective at detecting viruses,” Wu said.

The puzzle was to figure out a way to make the virus bigger.

The trick, he said, turned out to be relatively simple: just add water.

Similar to cloud formation, Wu’s device condenses water vapor on the tiny virus particles as they enter the sampler to increase their size and capture them more effectively.

Wu’s device also can catch any respiratory viruses, including the current influenza A virus subtype H3N2 (A/H3N2) and help scientists, engineers and medical researchers to identify which respiratory viruses exist and pinpoint proper measures for controlling their spread.

Once Wu held the key to the sampling puzzle, he began researching air samplers then on the market to see which one could best realize the goal. He teamed up with a company called Aerosol Dynamics.

Prototypes of their viable virus aerosol sampler are currently undergoing field testing at UF, Hong Kong University and Washington University in St. Louis.

Wu is now partnering with UF colleagues Professor John Lednicky in the department of environmental and global health, and Professor Hugh Fan in the department of mechanical engineering. Fan develops the devices to ensure that the captured virus particles can be analyzed on the spot, while Lednicky grows the viruses and assesses their infectivity, developing a plan to better protect the public from upcoming airborne viruses that season.

The samplers, Wu said, could be widely available in time for next year’s flu season.

T2Candida Shortens Time to Candida Detection vs. Culture

A small desktop blood test may improve the treatment of patients with candidemia by shortening the time it takes to detect Candida and identify the species responsible for infection compared with blood cultures, researchers found.

The T2Candida Panel (T2 Biosystems) was the first diagnostic test for candidemia approved by the FDA. Using MRI to scan blood, it can detect the five most common species of Candida: C. albicans, C. glabrata, C. parapsilosis, C. tropicalis and C. krusei.

In the DIRECT2 trial, which was funded by T2 Biosystems, Cornelius J. Clancy, MD, associate professor of medicine in the division of infectious diseases and director of the mycology program at the University of Pittsburgh, and colleagues evaluated the panel in patients who had been diagnosed with candidemia via a blood culture.

“Invasive Candida infections are common causes of mortality and morbidity in hospitalized patients. Blood cultures are the current gold standard for diagnosing invasive candidiasis, but we know they’re negative for Candida in 50% or more of bloodstream-disseminated candidiasis,” Clancy told Infectious Disease News.

From March 2012 through August 2013, Clancy and colleagues enrolled 152 patients at 14 hospitals who had been diagnosed with one of the five Candida species and performed follow-up blood testing using the T2Candida panel and companion blood cultures. The median time between the collection of the original diagnostic blood cultures and the retesting of the samples was 55.5 hours. Most (74%) patients had been given at least one dose of an antifungal agent prior to obtaining the follow-up sample.

T2Candida had a sensitivity of 89%, obtaining positive results in 32 of 36 patients with positive companion cultures, Clancy and colleagues reported. They also found that the panel performed similarly to companion blood cultures in patients with recent candidemia who were not receiving antifungal treatment, with positive results in 32.5% of cases compared with 30%. In patients who were receiving treatment, the panel was positive in 50% of patients, and companion blood cultures were positive in 21% of patients.

“The data demonstrate that T2Candida reliably diagnosed candidemia when collected at a time that blood cultures were positive,” Clancy and colleagues wrote. “T2Candida may improve the management of candidemia by shortening times to Candida detection and species identification compared to blood cultures, thereby facilitating more rapid institution of definitive antifungal therapy.”

According to Clancy, nonculture tests like T2Candida have the potential to identify cases that are currently missed by blood cultures.

“Moreover,” he said, “the turn-around time for T2Candida is typically about 5 hours, compared to several days for blood cultures. Therefore, clinicians can get both positive or negative results more rapidly. The time to initiation of an active antifungal drug is a crucial determinant of survival in patients with candidemia.”

Clancy said the test was in development “many years” before the emergence of C. auris, but that there is “nothing from a technical standpoint” that would prevent it from being able to detect the invasive and often drug-resistant fungus that has been compared to a superbug. – by Gerard Gallagher


Clancy CJ, et al. Clin Infect Dis. 2018;doi:10.1093/cid/cix1095.

Israel-Singapore Partnership Sires $5 Devices to Detect Dengue Fever, Strokes

BGN Technologies, the technology arm of Ben-Gurion University of the Negev, is developing a $5 device that can diagnose dengue fever and stroke symptoms. The device is being created in collaboration with Singapore-based firm Biosensorix.

The joint development is a biosensor that can detect the composition of chemicals in the blood to diagnose diseases within minutes, eliminating the need to send blood and tissues samples to the lab.

Immunosensors or biosensors are medical devices that measure the levels of chemicals in the bloodstream by detecting the electrical currents created due to interaction between enzymes. Based on the presence and quantity of specific chemicals, medical staff can determine the existence and severity of a disease.

While several kits exist in the market today that can be used at home or in clinics to diagnose diseases, the new kit is a “breakthrough” because it can quickly measure the level of a biomarker very cheaply, said Luka Fajs, CEO of Biosensorix.

“We are trying to decentralize medicine by allowing patients to test themselves at home and receive immediate test results at a very low cost to healthcare providers, eliminating the need for lab testing,” said Fajs.

The kit can diagnose dengue fever, a disease caused by a virus transmitted from mosquitoes most commonly found in Southeast Asia and South America. The quick diagnosis enables medical staff to start treating patients without needing to wait for lab results, the company says.

Currently in advanced prototype stage, the kit is made up of two components: a USB stick-like blood-testing strip which the patients use by pricking their finger to collect blood samples, and a second device that looks like an iPhone, into which they insert the USB with the blood sample to analyze its chemical composition.

“Most people with dengue fever can be released to home care, yet are kept at the hospital until results come in,” said Robert Marks, professor at the Avram and Stella Goldstein-Goren Department of Biotechnology Engineering at BGU and co-founder of Biosensorix. “With the new diagnostic kit, the physician can release the patient within half an hour, saving time and money.”

A second kit currently under development can provide information that would help prevent stroke and detect the possibility of a secondary stroke, which often occurs after the patient is released from the hospital, he said.

The diagnosis of a stroke — a condition caused by a blood clot that plugs blood vessels in the brain and prevents the flow of blood, resulting in the loss of brain cells within minutes — currently relies largely on clinical assessments and imaging procedures like MRIs.

The device, which can be used at home, has two applications for stroke. It can help prevent the occurrence of stroke by monitoring the patient’s blood chemistry on a weekly basis and providing risk assessments quickly, within 15 minutes, compared with lab testing which can take days or weeks, said  Fajs.

A second advantage is that in emergency situations, it provides medical professionals with valuable and timely information they can use to not only to speed up the process of diagnosis but also determine the best course of action.

“With a stroke, every minute counts. A quick quantitative test means rapid diagnosis that is necessary for accurate, timely treatment. This can save brain functions and even lives,” said Ora Horovitz, senior VP Business Development at BGN Technologies

The collaboration between the university and Biosensorix was born as part of the Singapore-Israel NRF CREATE program, which was founded in 2015 to fund research proposals based on cooperation between Israeli and Singaporean researchers.

Nearly $1 million in funding was provided by the Singapore government for the project.

“We received a boost of funding from Singapore which furthered the development of the intellectual property,” said Marks. “Singapore has an excellent infrastructure and is very much interested in developing startups, and we took advantage of these opportunities.”

The multinational partnership has paved the way for Israeli researchers to  develop medical technologies for diseases that are not native to the local market, such a dengue fever. These developments can later be scaled to large markets like Southeast Asia and South America, where the condition has a much larger prevalence, Marks said.

In recent years, an increasing number of giant tech companies like Google and Apple have made forays into the healthcare sector; the market in the US alone is estimated to be $3 trillion a year. As these healthcare platforms grow, they will need to rely on data gleaned from the smaller device manufacturers, said Fajs.

In 2015, Google Ventures, the investment arm of Alphabet Inc., was ranked as one of the leading investors in digital health, with nearly 31 percent of the company’s investment dollars spent on healthcare technologies,  according to Bloomberg.

Apple announced last month that its iOS 11.3 update includes a beta version of its Health app that will allow iPhone users to store and share their medical records from a range of healthcare systems in the US. Twelve hospitals and clinics have partnered with Apple for the pilot, including John Hopkins Medicine, Cedars-Sinai, and Penn Medicine.

Currently in the pre-series A funding stage, Biosensorix plans to introduce the devices to larger markets within two years after completing the initial manufacturing phase and receiving formal FDA approval, the company said.