Monday, April 11, 2016

Bruker Introduces Important MALDI Biotyper Enhancements at ECCMID 2016: New Disposable MBT Biotargets 96, MBT Subtyping and Carbapenem Resistance Tests

At the 26th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID), Bruker (Nasdaq: BRKR) introduces enhancements to the MALDI Biotyper (MBT) routine microbial identification workflow, and high-value expansions of the MBT assay menu to strain subtyping and to antimicrobial resistance testing for clinical and microbiology research.

1. MBT Biotargets 96 Further Improve Microbiology Laboratory Efficiency and Quality with Individually Bar-coded, Easy-to-Use and Time-saving Disposable MALDI Sample Targets

The disposable MBT Biotargets 96 further improve laboratory efficiency for the MALDI Biotyper workflow. They have 96 sample spots and can be disposed of after all spots have been utilized. The novel MBT Biotargets 96 use proprietary AnchorChip™ technology for exact sample positioning and maximum sensitivity. The targets are individually bar-coded, providing sample traceability for each MALDI Biotyper run, and their unique material offers optimal flatness and product integrity. The MBT Biotargets 96 are now available for European clinical customers, following IVD directive EC/98/79.

2. Rapid, New MBT Subtyping Module for Research Use Only (RUO)

The MBT Subtyping Module enables fast detection of selected antibiotic-resistant strains in Bacteroides fragilis and Staphylococcus aureus isolates. Identification and resistance detection is performed in a single automated analysis, providing species identification followed by selected resistance information. For B. fragilis, as the most prevalent species of the Bacteroides genus, the MBT Subtyping Module automatically detects carbapenemase producing strains that carry the cfiA gene.

For S. aureus the MBT Subtyping Module detects strains carrying the mecA cassette expressing the PSM protein. For this resistance mechanism, the new test can differentiate MRSA (methicillin-resistant Staphylococcus aureus) from MSSA (methicillin-sensitive Staphylococcus aureus) within minutes.

3. Rapid, New MBT Selected Test of Antibiotic Resistance (STAR) for Beta-lactamase (BL)

The new MBT STAR-BL assay supports the generic detection of Gram-negative carbapenemase producers, using a patented, functional beta-lactamase test for selected antibiotics. The MBT STAR-BL software (RUO) monitors mass shifts introduced by enzymatic degradation of betalactam antibiotics, such as penicillins, 3rd- generation cephalosporins and carbapenems. The MBT STAR-BL test consists of a software module for easy data analysis and the MBT STAR-BL IMI kit, which is currently in performance evaluation studies. During the evaluation, this kit is for investigational-use-only (IUO), and available for participating collaboration partners.

Professor Jens Jorgen Christensen at the Department of Clinical Microbiology at Slagelse Hospital in Denmark stated: "Carbapenemase producing enterobacteria strains are on the rise globally. Rapid recognition in risk situations is desirable for optimal antibiotic treatment of patients. The MBT STAR-BL IMI approach will be helpful for that purpose, as increased resistance can be detected in less than three hours."

Philip Perry, the Business Development Manager for Consumables at Bruker Daltonics, added: "Bruker is committed to the continuous expansion of our MALDI Biotyper platform, e.g. with enhancements of our reference libraries, increased customer convenience and expansion of the MALDI Biotyper assay menu, in order to further increase workflow ease-of-use and efficiency, and now also for rapid testing of selected antibiotic resistance mechanisms in clinical research."

New Study Data Show OpGen’s QuickFISH Blood Culture Pathogen ID Tests to be Rapid and Cost-Effective, with Utility to Inform Antimicrobial Stewardship Decisions

OpGen, Inc., a precision medicine company using molecular diagnostics and bioinformatics to combat infectious disease, today announced that new study data on its QuickFISH® tests were presented at the 26th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID) being held from April 9th to 12th in Amsterdam.

The studies were conducted at the NHS Royal Free Trust Hospital in London and were presented in two posters on Saturday, April 9th.
  • Abstract #2455. Rapid differentiation of Staphylococcus aureus from coagulase-negative staphylococci directly from positive blood cultures: prospective comparison of four rapid methods; Rebecca Gorton, et al.  
  • Abstract #6601. The potential utility of QuickFISH on positive blood cultures to inform antimicrobial stewardship decisions; Tehmina Bharucha, et al.
In the first poster, investigators concluded that QuickFISH rapidly differentiated Staphylococcus aureus from coagulase-negative staphylococci directly from blood cultures. It was the fastest and most cost-effective nucleic acid method evaluated with a time-to-result of 20 minutes, and had a higher diagnostic concordance to routine methods than MALDI-TOF. Other methods evaluated included Cepheid GeneXpert® and a tube coagulase test.

In the second poster, the investigators reported that communicating Gram stain and QuickFISH results together with clinical context (including local resistance patterns) can help inform physicians in early patient management decisions.  They concluded that the development of a treatment algorithm based on the results may enhance the clinical benefits of QuickFISH.

Also at the conference, OpGen previewed the QuickFISH Digital Imager, which will automate the QuickFISH procedure and digitize the results.  The bench-top instrument is currently in development and is being designed to eliminate the requirement of a fluorescence microscope and darkroom.  Development plans include the integration of the Digital Imager with Acuitas Lighthouse™, its bioinformatics platform that will allow rapid analysis of pathogen identity in the context of an antibiotic resistance database, providing decision support for patient management.

“We are encouraged by the impact we are making with premier healthcare institutions such as the NHS Royal Free Trust Hospital with our current QuickFISH products, and are equally excited about our future and the progress we are making with our development programs,” commented Kevin Krenitsky, M.D., OpGen’s president.  “We are working to transform the way infectious diseases and antibiotic resistance are managed through the use of molecular and digital technologies. Our aim is to apply the principles of precision medicine to improve patient outcomes, reduce hospital costs and preserve the supply of effective antibiotics.”

New Assay Offers Improved Detection of Deadly Prion Diseases

Transmissible spongiform encephalopathies (TSEs), or prion diseases, are a family of rare progressive, neurodegenerative illnesses that affect both humans and animals. TSE surveillance is important for public health and food safety because TSEs have the potential of crossing from animals to humans, as seen with the spread of mad cow disease, or bovine spongiform encephalopathy (BSE). A study in The Journal of Molecular Diagnostics describes an advanced assay that offers better sensitivity than currently available tests for detecting a prion disease affecting elk.

"The significance of TSEs on human health was not entirely realized until cases of variant Creutzfeldt-Jakob disease (vCJD) in humans had been discovered in the years following the BSE outbreak in the United Kingdom. These vCJD cases were associated with consuming meat products contaminated with BSE prions," explained lead investigator Stefanie Czub, DVM, PhD, of the Canadian BSE Reference Lab, Canadian Food Inspection Agency, Lethbridge Laboratory, Lethbridge, Alberta (Canada). "The result is that many countries have enacted TSE surveillance programs, aiming to eradicate livestock-related TSEs."

Surveillance programs rely on highly sensitive diagnostic methods to detect infections early. Addressing the need to define steadfast analytical performance criteria for prion amyloid seeding assays (ASAs), researchers developed a method to measure prion protein conversion time (from normal cellular form to prion form) by a combination of statistical analyses to obtain a prion-detecting ASA with a known degree of confidence. They compared the sensitivity of the new assay technique, the timed prion seeding assay (tASA), to other currently available tests (two bioassays in laboratory rodents and three commercially available TSE rapid tests).

The test samples came from elk brains infected experimentally with chronic wasting disease (CWD), a prion disease that affects cervids (hoofed ruminant mammals in the deer family). The investigators were able to define clear cut-off criteria, allowing determination of TSE-positive and TSE-negative states. Unlike TSE rapid tests, ASAs also have the potential to detect and measure TSE infection in blood, saliva, or urine. This would offer clinical advantages, such as the ability to sample blood instead of relying on more invasive tissue biopsy and to screen blood donations for contamination.

"We found that the tASA was at least as sensitive as two rodent bioassays and up to 16 times more sensitive than three different TSE rapid tests," noted lead author John G. Gray, MS, Canadian BSE Reference Lab, Canadian Food Inspection Agency. "This study should further advance ASAs as recognized prion detection systems. We believe this methodology represents the future for prion diagnostics, especially concerning human health, for example in screening blood donations."

The tASA is an in vitro method that mimics the conjectured mechanism of prion propagation in vivo. It is a conversion assay that uses recombinant prion-related protein as a substrate and detects conversion via changes in fluorescence. The report describes time specifications for the assay to help avoid false-positive results (30 hours) or false-negative results in weakly-positive samples (48 hours), as well as the number of replications necessary for adequate sensitivity (2-12).

"This study represents an important first step for the tASA diagnostic protocol to gain regulatory approval for its use in TSE surveillance programs targeting CWD in cervids," commented noted authority Holger Wille, PhD, of the University of Alberta Department of Biochemistry and Centre for Prions and Protein Folding Diseases in Edmonton, Alberta (Canada). "Additional work will also be needed to fine-tune and test tASA for the detection of prions in peripheral organs and environmental samples, which represent a substantial unmet need to track the spread of CWD prions among North America cervids as well as in the environment."

Prions are abnormal, transmissible pathogenic agents that induce abnormal folding of specific, normal cellular proteins. Because these proteins are concentrated in brain tissue, brain damage is characteristic of prion diseases. Prion diseases generally progress rapidly and are associated with high mortality. Prion disease occurs when the normal cellular form of prion-related protein converts or conformationally changes to the disease form. Once the disease form is introduced, it becomes self-perpetuating as it converts the normal form into more of the disease form. CWD is currently spreading throughout the U.S. and Canada, is also present in South Korea, and was just diagnosed in a first reindeer in Norway.


"Defining and Assessing Analytical Performance Criteria for a TSE-Detecting Amyloid Seeding Assay," by John G. Gray, Catherine Graham, Sandor Dudas, Eric Paxman, Ben Vuong, and Stefanie Czub (DOI: Published online in advance of The Journal of Molecular Diagnostics, Volume 18, Issue 3 (May 2016)

Sunday, April 03, 2016

WHO Calls for Early Diagnostic Tests for Lassa Fever

Lassa fever has killed more than 160 people in West Africa, most of them in Nigeria, since November 2015. Many of these lives could have been saved if a rapid diagnostic test were available so that people could receive treatment early.

Since November 2015, Nigeria, Benin, Sierra Leone and Togo have reported more than 300 cases of Lassa fever and 164 deaths. Nigeria accounts for the majority of the cases with 266 cases and 138 deaths reported in 22 of the country’s 34 provinces. Benin has recorded 51 cases and 25 deaths, Togo and Sierra Leone each reported two cases.

Lassa virus is carried by the Mastomys rat, which is found in parts of West Africa. The virus is transmitted to humans from direct contact with infected rats by catching and preparing them for food, or through contact with food or household items contaminated with rat feces or urine. The virus can also be transmitted through contact with an infected person’s body fluids.

Around 80 percent of people who become infected with Lassa virus have no symptoms or they have symptoms that mimic other illnesses, such as malaria, making it difficult to treat them. Symptoms include fever, fatigue, nausea, vomiting, diarrhea, headaches, abdominal pains, sore throat and facial swelling.

“Without early diagnosis and treatment, 1 in 5 infections result in severe disease, where the virus affects several organs such as the liver, spleen and kidneys,” explains Dr. Pierre Formenty, an expert in hemorrhagic fevers at the World Health Organization (WHO). “We need resources to invest in diagnostics to easily, accurately and safely test for Lassa fever as we do for malaria and HIV. Without a proper diagnosis, many people do not receive the correct treatment and that is why we see so many people with Lassa fever dying each year.”

Ribavirin has been used successfully in the treatment of confirmed Lassa cases. This drug can treat infected people if it is administered as soon as the first signs appear.

The first known case of Lassa infection outside of Africa has been reported in Germany. One person, a funeral home employee was infected after direct contact with an American who died of the disease in February 2016. The American was a medical director of a missionary hospital in Togo who was evacuated to Germany where he died.

“This is the first time that secondary transmission of the infection is reported in Europe,” says Formenty. “The risk for further transmission of Lassa fever in Germany and also West Africa is low and limited to hospital settings caring for the cases, with all contacts accounted for and monitored.”

WHO continues to monitor the epidemiological situation and conduct risk assessments based on the latest available information.

Fujifilm and FIND* Sign Development Contract to Develop Highly Sensitive Rapid Tuberculosis Diagnosis Kit for Developing Countries

FUJIFILM Corporation has signed a joint development contract with the Swiss non-profit organization FIND (Foundation for Innovative New Diagnostics) for the development of a highly-sensitive rapid tuberculosis (TB) diagnosis kit. The kit uses an immunochromatography*** method which applies silver halide amplification technology*4. FIND is an organization dedicated to advancing the development and adoption of new diagnostic technologies for infectious diseases that are suited for developing countries. Under this contract, FIND will support Fujifilm in the development of a convenient, rapid, low-cost TB diagnosis kit with high diagnostic capability for developing countries. This partnership has been selected for a subsidy by the Global Health Initiative Technology (GHIT) Fund, a fund for the development of innovative new treatments, vaccines, and diagnostic agents in Japan, receiving approximately ¥216 million. The timeline of development is from April 2016 through October 2017, for one year and seven months.

TB is one of the world's major infectious diseases*5 infecting 9.6 million people, and causing the death of 1.5 million people*6 annually around the world. Of all TB patients, 86% are concentrated in developing countries in Africa and Southeast Asia*7. TB infections have a serious impact on the societies and economic activities of these countries due to the ongoing spread of the disease and the cost of medical treatment for patients. HIV is also common among populations in developing countries, and when people contract HIV, their immune systems are adversely affected, making HIV patients approximately 30 times more likely than healthy individuals to develop TB. Patients who have both TB and HIV frequently become critically ill, with one in three cases of such co-infections leading to mortality. Therefore, it is important to provide regular TB screening for people living with HIV in order to begin TB treatment at an early stage of the disease.

In developing countries, the diagnosis of TB is frequently carried out using a microscope to search a patient's sputum for the presence of the bacteria that causes TB (Mycobacterium tuberculosis). However, one issue with this method of diagnosis is that accurate results cannot always be obtained due, for example, to failure to identify the TB bacteria or due to extrapulmonary tuberculosis, which is caused by the presence of TB-causing bacteria outside the lungs. In recent years, genetic analysis of sputum, which is highly reliable, has gradually become more widespread, but this method requires laboratory technicians with specialist techniques and expensive diagnostic equipment, in addition to a steady electricity supply. Therefore, the development of an alternative diagnostic method better suited for developing countries is a pressing issue. Furthermore, it is hoped that specimens other than sputum can be used for diagnosis since there are certain patients from whom it is difficult to obtain sputum, such as small children and elderly persons, and also because sputum diagnosis is not effective in the case of extrapulmonary TB, which affects numerous HIV patients.

Fujifilm and FIND have focused on lipoarabinomannan (LAM)*8, a compound specifically produced by TB that is excreted in the urine. By using Fujifilm's highly sensitive virus detection technology, the silver halide amplification technology, Fujifilm and FIND will work to develop a kit suitable for resource-poor countries that will allow the prompt identification of the presence of the TB bacteria simply by placing a urine sample into a cartridge in a device that does not require electricity. The partners aim to develop a kit that can serve as a primary diagnostic tool for TB, including in HIV patients.

Fujifilm has already developed the highly sensitive immunochromatography influenza diagnosis system for identifying the influenza virus, and this was launched in Japan in October 2011. The system uses two different types of antibodies to identify the presence of a virus in the sample and, through simple operations, the results can be obtained in 3.5 to 15 minutes. By making use of the silver halide amplification technology used to develop photographs, this system achieves approximately 100 times the sensitivity of standard diagnostic agents*9. This proprietary technology, which allows the detection of very small volumes of influenza virus in the early stages of infection, has been highly appreciated and is being introduced at many medical institutions in Japan. Furthermore, Fujifilm is working to apply the technology to the detection of non-influenza infections such as adenovirus and the RS virus.

Fujifilm perceives solving social issues as an opportunity for growth in the medical business. Fujifilm will continue to proactively promote research and development to expand its operations and contribute to the development of medical care around the world, as well as maintain and promote public health through innovative products.

New Real-time PCR Assay Offers Validated Method for Rapid Detection of Cronobacter species in Powdered Infant Formula

To help safeguard against release of infant formula contaminated with Cronobacter bacteria, which can produce fatal effects on newborn and premature babies, a new simple-to-use PCR assay enables manufacturers and their suppliers to get fast and accurate test results on powdered infant formula, with and without pro-biotics, and on manufacturing environmental process samples.

The new Thermo Scientific SureTect Cronobacter Species Real-Time PCR Assay delivers PCR results in as few as 18 hours following sample receipt. This enables infant formula manufacturers and their suppliers to process and evaluate samples quickly. The assay has been granted NF VALIDATION certification by AFNOR Certification, and can form part of a test workflow that helps customers comply with the Commission Regulation (EC) No 2073/2005 on microbiological criteria for foodstuffs.

“We are proud to be able to help our customers protect this vulnerable consumer group with a test method that is both highly accurate and fast compared to the ISO Standard method for detecting this organism,” says David Panneton, vice president commercial operations, microbiology, Thermo Fisher Scientific. “The new SureTect Cronobacter Assay enables our customers to take appropriate action, including fast release of product to market, which is a primary consideration for today’s pathogen test selection.”

Looking at the Bacteria Inside: A New Method of Viewing TB Bacteria

Although tuberculosis (TB) is commonly thought of as being a disease that mainly affects nineteenth century poets and Victor Hugo characters, it is still the second-most common cause of mortality from an infectious disease in the world, killing nearly three people every minute. Every March 24, on World TB Day, the global health community recognizes the work of Robert Koch, who announced on that date in 1882 his discovery of Mycobacterium tuberculosis, the bacteria that causes TB.

TB is generally a curable disease—but first it must be diagnosed, and that has been one of the biggest barriers in its elimination. M. tuberculosis grows very slowly, so traditional methods of diagnostic testing take a very long time. A team from the Texas A&M Health Science Center College of Medicine has been working on a new approach. The work is led by Jeffrey Cirillo, Ph.D., a professor in the Department of Microbial Pathogenesis and Immunology.

Cirillo’s previous research, which has been supported by the Bill and Melinda Gates Foundation and the Wellcome Trust, has focused on developing a rapid yet inexpensive test for tuberculosis. He and his team have nearly perfected the technology and are in the process of bringing the test to market.

The slow growth of M. tuberculosis also makes development of new therapeutics difficult because it takes so long to determine if the treatment is working.

Now, Cirillo and his team have developed a method, which they reported recently in the journal PLoS ONE, of detecting whether a potential treatment is working as early as a day after it is given. “Since M. tuberculosis takes about a month to quantify using traditional approaches, any method that allows direct determination of the amount of bacteria present is very valuable and speeds progress, saving months every time an experiment is done,” Cirillo said. “Our methods allow this.”
Using optical imaging technology, the researchers can observe the bacteria in real time, in living animals, without harming them. This way, it is easy to determine if a proposed treatment is working and the number of bacteria is decreasing.

“There have never been sensitive enough systems to measure the bacteria directly in animals,” Cirillo said. “This is a completely new technology and has nearly limitless applications to microbiological research, particularly in animals, but it increases sensitivity in any experimental system.”

Drug-resistant TB, now present everywhere in the world, is hampering efforts to fight the disease because the usual drugs used to treat it are no longer effective. Therefore, new, more effective treatments need to be developed.

The researchers first had to find the best fluorescent protein to help them visualize the bacteria.
“We use very sensitive systems that can actually see the signal through mammalian tissue,” Cirillo said. “This works best in the near-infrared, which is where our signals are primarily produced.”
Active TB causes symptoms such as cough, fever, night sweats, weight loss. If left untreated—a common scenario in developing countries lacking health care infrastructure—a person with active TB has only a 50 percent chance of survival, and he or she will infect an average of 10 to 15 people each year.

“The goal in TB research is complete eradication of the disease,” Cirillo said. “We think this new technology is one tool to do just that.”

Citation: R21HL115463; R01AI104960; 48523; PLoS ONE