Monday, November 21, 2016
The innovative aspect of the "Origami Diagnostics to Accelerate Malaria Elimination" project is its use of "paper microfluidics" to produce a malaria test that is one hundred times more sensitive than commercially available tests while retaining the low cost and simplicity required for real world application.
"In order to eradicate malaria, we must be capable of detecting the individuals that carry the malaria parasite but don't show any symptoms," said Wright. "Current commercial malaria tests are not capable of doing this. That is why we have engineered our origami test with the sensitivity required to identify these individuals. At the same time, we have designed it so it will be extremely inexpensive to make and so it will be as easy to operate as current lateral flow detectors like the pregnancy test."
One of the factors that limit the sensitivity of current rapid malaria tests is the small amount of blood that they can process: a single drop. To address this problem, the origami detector consists of a relatively large well that is capped with a porous membrane. The membrane has a special coating that selectively catches proteins produced by the malaria parasite, called biomarkers. Users dilute a few milliliters of blood in a special liquid and pour it through the membrane, which snags the biomarkers as the mixture percolates through. This allows it to collect a much larger number of biomarkers than commercial tests. Once that is done, the users detach the well and throw it away. Then they fold the membrane onto an attached sheet of paper printed with special inks. These inks contain sensor molecules that change color when they bind with parasite biomarkers. Next they wet down the membrane/paper sandwich with a special liquid that releases the biomarkers from the membrane so they will come in contact with the sensor molecules, causing them to change color.
To receive funding, Wright and other Grand Challenges Explorations winners described a "bold idea" in a two-page online application in one of six critical global heath and development topic areas.
FDA Completes Transfer of Emergency Use Authorization for ReEBOV® Ebola Antigen Rapid Test to Zalgen Labs
The test is to be used for the presumptive detection of Ebola viruses (detected in the 2014 – 2016 West Africa outbreak) in individuals with signs and symptoms of Ebola virus infection in conjunction with epidemiological risk factors (including geographic locations with high prevalence of Ebola infection). The authorized ReEBOV Antigen Rapid Test is intended for circumstances when the use of a rapid Ebola virus test is determined to be more appropriate than use of an authorized Ebola virus nucleic acid test.
Ebola virus is indigenous to western and central Africa and is one of the deadliest viruses in the world, with mortality rates of between 30 and 90 percent. The ReEBOV Antigen Rapid Test for Ebola was the first rapid diagnostic test (RDT) and the first immunoassay authorized for emergency use by the FDA for the presumptive detection of Ebola virus, and also the first listed for procurement by the World Health Organization (WHO) under the Emergency Use Assessment and Listing procedure. Under the terms of a previously announced collaboration agreement, NOWDiagnostics Inc. (NOWDx) will manufacture the ReEBOV Antigen Rapid Test for Ebola virus as well as other Zalgen diagnostic products.
Unlike molecular testing, which in West Africa can still take days to return results from central testing laboratories, the Zalgen ReEBOV RDT is a point-of-care test that can be used in laboratories or facilities adequately equipped, trained and capable of such testing. That includes testing in treatment centers and public health clinics or in field laboratories with trained personnel capable of such testing. Instead of taking days for lab results, the ReEBOV RDT uses a drop of blood from a finger prick to deliver a presumptive detection of Ebola virus antigen in as little as 15 - 25 minutes, potentially allowing trained public health workers to rapidly screen, isolate and initiate care of suspect Ebola patients. Medical personnel will be able to quickly identify hotspots and may prevent resurgence of cases in future outbreaks.
The ReEBOV Antigen Rapid Test was developed in cooperation with additional members of the Viral Hemorrhagic Fever Consortium (VHFC), a collaboration of academic and industry members headed by Tulane University, including Autoimmune Technologies LLC, The Scripps Research Institute and the University of Texas Medical Branch at Galveston, as well as other collaborators in West Africa, particularly the Ministry of Health and Sanitation (MOHS) of the Republic of Sierra Leone and the medical personnel of the Kenema Government Hospital in Kenema, Sierra Leone, a number of whom died fighting the 2014-16 Ebola outbreak.
“This emergency use authorization from the FDA enables Zalgen and our distribution partners to continue providing this remarkable product worldwide to test suspected Ebola cases,” said Zalgen Managing Director, Luis Branco, PhD. “Zalgen and the VHFC are already working with NOWDx to develop next generation diagnostic tests for Ebola, Lassa and other hemorrhagic fevers as well as other tropical diseases.”
Development of the ReEBOV Antigen Rapid Test for Ebola was supported by the National Institute of Allergy and Infectious Diseases (NIAID) of the National Institutes of Health (NIH) (grants 1R43AI088843 and 2R44AI088843). Additional support was provided by The Bill & Melinda Gates Foundation and the Paul G. Allen Family Foundation.
Under the terms of the agreement, Merck will provide access to its archive of over 200,000 bacterial pathogens gathered over the last 15 years through the Study for Monitoring Antimicrobial Resistance Trends (SMART), one of the world’s largest surveillance studies of antimicrobial resistance supported by Merck in collaboration with International Health Management Associates (IHMA). OpGen will perform genomic analysis, microbiology testing for drug resistance, and incorporate this information into its Acuitas® Lighthouse Knowledgebase and the development of rapid DNA tests. This new molecular testing and informatics approach is being developed to help transform antibiotic decision making for doctors managing acute care patients with blood, respiratory, urinary tract, and soft tissue infections. In addition to identifying resistance determinants to predict antibiotic failures, the OpGen technology is being evaluated as the foundation for utilizing molecular diagnostic tests to predict pathogen susceptibility and guide patient management choices to improve patient outcomes.
“This collaboration builds upon the promise of our DNA-based genetic tests, Lighthouse Knowledgebase and antibiotic resistance decision making tools to make a significant impact on hospital infections,” said Evan Jones, chairman and CEO of OpGen. “Access to Merck’s SMART surveillance network data has the potential to greatly accelerate our internal development efforts in validating our rapid diagnostic tools and bolster data acquisition for our Lighthouse Knowledgebase.”
Recent studies have indicated that antimicrobial resistant infections currently claim 50,000 lives each year across the United States and Europe alone, with many hundreds of thousands more dying in other areas of the world. In September, world leaders at the United Nations called rising antimicrobial resistance a fundamental threat to human health, development, and security. As a result, for the first time, Heads of State committed to taking a broad, coordinated approach to combat rising antimicrobial resistance including the development of new medicines and rapid diagnostics.
“Rapid diagnostics for pathogen identification and antibiotic susceptibility testing are central to developing global solutions for antimicrobial resistance,” said Dr. Eliav Barr, senior vice president, Infectious Diseases and Vaccines Clinical Development, Merck Research Laboratories. “By providing OpGen with access to our archive of bacterial pathogens, we hope to expedite the development of rapid diagnostic tests and enable prompt and informed antibiotic prescribing to improve patient outcomes.”
OpGen will initially perform molecular analyses on up to 10,000 pathogens to identify markers of resistance to support rapid decision making using the Acuitas Lighthouse® MDRO Management System (“Lighthouse Portal”), and to speed development of OpGen’s rapid diagnostic platforms. OpGen’s Lighthouse Portal and Knowledgebase are being developed to provide antibiotic stewardship and tracking information for drug resistant pathogens in hospitals and health systems. Merck will gain access to the high-resolution genotype data for the SMART isolates as well as access to OpGen’s Lighthouse Portal to support internal research and development programs.
About the SMART Study
The Study for Monitoring Antimicrobial Resistance Trends (SMART) was initiated by Merck in 2002 to monitor the in vitro susceptibility of clinical isolates to 12 commonly used antibiotics in different regions of the world to survey changing trends in antibiotic susceptibility. SMART currently monitors antibiotic activity against gram-negative bacteria isolated from two common types of infection: intra-abdominal and urinary tract infections. Isolates have been collected from patients with complicated intra-abdominal infections since 2002 and from patients with complicated urinary tract infections since 2010. In 2016, Merck anticipates that more than 41,000 isolates will be collected.
This research agreement is executed between OpGen and Merck Sharp & Dohme Corp., a wholly-owned subsidiary of Merck & Co. Inc. The Merck Global Health Innovation Fund, a venture capital arm of Merck, is an investor in OpGen Inc.
During the AVS 63rd International Symposium and Exhibition being held November 6-11, 2016, in Nashville, Tennessee, Stephane Evoy, an applied physicist from the University of Alberta, will explain how the team recognized the limited reliability of antibodies in providing bacteria detection with specificity. Instead they used phage-derived proteins, proteins developed from the bacteria-invading viruses, for detection of pathogenic bacteria to address this deficiency. This work has implications not only in disease diagnosis, but also in food and water safety.
"The high specificity of phages offers a potent alternative for the targeting of pathogens," Evoy said. "More specifically, recombinant phage-receptor-binding proteins (RBPs) responsible for phage-host specificity can be used as biological probes and present numerous advantages over the use of a whole phage."
The study used skim cow milk spiked with different phages or combinations of phages, such as mycobacteria (MAP) and Escherichia coli cells, and a unique process to capture the DNA after incubation. The entire process took less than 24 hours and resulted in significantly better sensitivity of detecting targeted DNA.
"The use of phage-derived proteins in such a manner was quite unique when we started that work back in 2005, but since then the approach thrived, and multinational companies integrated this into their product line," Envoy said. "However, there is still a lot of work to be done in terms of applying the technology to diseases such as tuberculosis and staphylococcus infections."
In addition to demonstrating this capture technique, the research team designed and developed a sophisticated bacteria detector comprised of an array of microresonators, able to enumerate bacteria over a large area and detect the attachment of a single cell anywhere on the array. The devices were prepared with their phage proteins, adding this high specificity of detection to the spatial precision offered by the array design.
"We are looking forward to adapting this technology for the rapid diagnosis of drug-resistant bacteria," Evoy said. "It could go a long way toward make microbial testing methods both more rapid and affordable."
The OSOM® Ultra Flu A&B Test delivers fast, accurate results in 10-15 minutes by utilizing a simple procedure across multiple sample types, including nasal swabs, nasopharyngeal swabs, and in moderate settings, nasopharyngeal aspirate/wash. The performance of the OSOM® kit by users at CLIA waived sites was measured against the preferred standard of polymerase chain reaction (PCR), with a Positive Percent Agreement of 89.2% for Flu A and 86.4% for Flu B and Negative Percent Agreement of 99.4% for Flu A and 99.0% for Flu B.
According to the CDC Foundation, there are approximately 31.4 million outpatient visits and 200,000 hospitalizations each year related to influenza. An estimated 20 million rapid tests have been performed in the U.S. each year since 2013, with a projected 4.5% CAGR through 2019. ]2,3]
CLIA waiver designation will enable healthcare providers to perform the OSOM® Ultra Flu A&B Test using nasal or nasopharyngeal swabs, to deliver simple and accurate test results in locations such as the emergency room, community-based settings, urgent care and physician offices. By using the OSOM® Ultra Flu A&B Test healthcare practitioners will provide patients with a rapid, accurate diagnosis while on site, thereby helping prevent unnecessary antibiotic prescriptions, reduce hospitalization, and minimize influenza transmission. 
"The excellent performance benefits of the CLIA Waived OSOM® Ultra Flu A&B Test will aid in the fast, accurate detection of influenza through an easy to use product that provides physicians and patients the convenience of rapid results and treatment in one visit," says Robert Schruender, President and Chief Operating Officer of Sekisui Diagnostics. "The test complements our other OSOM® CLIA waived rapid diagnostic tests and is another example of meeting our promise to our customers to deliver the products they need."
Along with influenza, Sekisui Diagnostics provides a broad line of OSOM® rapid tests for Strep A, Mononucleosis, Trichomonas, Bacterial Vaginosis, Helicobacter pylori, hCG and Fecal Occult Blood. Sekisui's expanding Point-of-Care product line also includes the FastPack® IP System, which offers a convenient, rapid, decentralized testing solution for Vitamin D, Testosterone, PSA, Free PSA, TSH, Free T4, and hCG.
Point-of-Care Testing (POCT) is a highly relevant and growing area of diagnostics, particularly due to the increased focus on cost efficiency through rapid and appropriate treatment. Market analysis and consulting firm Enterprise Analysis Corporation estimated the global market for POC testing at $5,924 million in 2015. 
2. GHX data
3. Kalorama 2014 Physician Office Laboratory Markets
4. Theocharis, George.Vouloumanou, Evridiki et al."Evaluation of a direct test for seasonal influenza in outpatients", European Journal of Internal Medicine, Vol. 21(2010) 434-438.
5. Enterprise Analysis Corporation, IVD Market Research 2016.
The new assay follows the path MetaSystems and Miacom Diagnostics have chosen and will be initially available to be used on the Metafer RPI Platform allowing an automated, high-throughput slide analysis directly after the samples are prepared using Miacom’s standard Direct Multiplex Imaging procedure. The MRSA/MSSA differentiation will be available as either a stand-alone assay for screening species of Staphyloccus for MRSA or as a combined multiplex assay detecting more than 10 important Gram positive and Gram negative bacteria in a sepsis. As all of miacom’s assays, this assay relies on the FDA-approved Direct Multiplex Imaging (DMI) technology which allows to test patient materials directly without the need for amplification in a multiplex fashion differentiating up to 14 pathogens in one single test. The identification of the bacterial strains is made possible via detection of fluorescent light given by the molecular probes used when they find their specific target molecules.
The launch of this addition to Miacom’s existing test kits is expected to take place in 2017 and will offer physicians to improve their treatment strategies. ”In the long run this will result in the reduction of the use of broadband antibiotics, reduce patient morbidity and mortality and also help clinics to save time and resources”, stated Dr. Mirko Stange, CEO Miacom Diagnostics.
Naturally occurring in freshwater lakes and rivers, the Legionella bacterium is harmless in small enough quantities, but problems start when it multiplies in plumbing systems, air conditioning units, Jacuzzis, decorative fountains or in a public water supply. Here it can be transmitted to humans when it condenses into droplets of fine mist which are inhaled and then settle in the lungs.
Roughly 5,000 cases are reported in the United States every year, while 2013 saw 5,851 cases reported by 28 EU Member States and Norway, according to the European Centre for Disease Prevention & Control (ECDC).
The European group POSEIDON, (or ‘Plasmonic-based automated lab-on-chip sensor for the rapid in-situ detection of Legionella’) intends to change all this, having developed their scanner to spot the deadly Legionella bacteria in under one hour, a process that normally takes 10 days of cultivation and analysis.
Equipped with tiny sensors, the device works by using the photonics technique of Surface Plasmon Resonance (SPR), a procedure that reads information from a refracted laser beam, allowing fast, highly sensitive, inexpensive detection from a small sample without the need for ‘labelling’, the process of binding to a protein in order to be detected.
SPR occurs when polarized beams of light hit a metal film at the interface of two media. A charge density oscillation of free electrons (or “surface plasmons”) at the metal film occurs, reducing the intensity of reflected light. The scale of the reduction depends on the substance on the metal at the interface. Information then gathered from the refracted can then be analysed, and a pre-programmed pathogen confirmed, resulting in an unambiguous detection of the bacteria in situ.
Surviving and flourishing at temperatures between 25º to 45º C, Legionella bacteria are normally prevented by heating water units above 70º C in order to kill them off. However new bacteria can form quickly, and not all of the pathogens are necessarily removed. The POSEIDON project aims to remove the uncertainty involved. Scientific coordinator, Roberto Pierobon explains:
“POSEIDON is a first for detecting Legionella with light and provides an inexpensive, user-friendly, state of the art early warning system on an air-conditioning unit. We aim to reduce the time involved in a diagnosis from 10 days to less than 1 hour. In order to prevent outbreaks at critical times of the year, we should be talking about a matter of minutes, rather than days.”
“Cells remain intact throughout the whole fluid transportation system in the device, and do not adhere to the fluidic piping and microfluidic channels. Virtually all of the bacteria cells in the sample are delivered to the sensing unit, giving extremely high sensitivity and specificity,” said Pierobon.
Hoping to have these revolutionary new pathogen detectors ready within 3 years, Bruno Bellò, project coordinator and CEO of Clivet, is excited about the implications for the future,
“The exciting feature of this device is that with future development, it could be recalibrated to look for other pathogens, which would provide incredible safety options for the environmental, medical or food industries,” Bellò said.
Earlier last year the POSEIDON consortium received funding of €4,068,781 from the Photonics Public Private Partnership, via the European Commission’s H2020 program for a three year research project. Coordinated in Italy, POSEIDON is comprised of a number of European partners, including Protolab, Clivet, A.R.C (Italy), Catlab (Spain), Metrohm Applikon (Netherlands), and Uppsala University(Sweden).