DNA-based PCR Test Accurately Predicts Antibiotic Resistance

A rapid high-throughput PCR test developed by OpGen “accurately and reliably” predicted antibiotic resistance in common bacteria at a rate of 90% or higher, researchers reported.

According to a study published in Antimicrobial Agents and Chemotherapy, the researchers evaluated more than 7,500 highly antibiotic-resistant clinical isolates of Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Pseudomonas aeruginosa for antibiotic-resistance genes and compared them with phenotypic resistance across penicillins, cephalosporins, carbapenems, aminoglycosides, trimethoprim-sulfamethoxazole, fluoroquinolones and macrolides.

“In the most comprehensive study of its kind, OpGen’s informatics technology (Acuitas Lighthouse) rapidly predicted antibiotic resistance to 15 antibiotics accurately and reliably, with an average positive predictive value of 90%,” G. Terrance Walker, PhD, senior vice president of research and development at OpGen, told Infectious Disease News. “On a larger scale, the paper supports a potential breakthrough in the global effort to speed up the rapid detection of antimicrobial resistance.

“Today, phenotypic methods based on bacterial growth are the standard of care. It can take days to provide results to patients. However, DNA testing can be completed in just hours and represents an attractive potential solution. Showing that DNA testing can be used to accurately predict antibiotic resistance has been a critical challenge that scientists have been working on for many years. This paper demonstrates that a well-defined set of DNA markers can potentially be used to perform rapid, accurate antibiotic resistance detection in key pathogens.”

Walker and colleagues reported testing 2,919 E. coli isolates, 1,974 K. pneumoniae isolates, 1,150 P. mirabilis isolates and 1,484 P. aeruginosa isolates, of which 34% originated in hospitals in North America, 23% in Europe, 13% in Asia, 12% in South America, 7% in Africa and 1% in Oceania, according to the study. Another 9% of tested isolates were of unknown origin.

According to the study, they “developed statistical methods to predict phenotypic resistance from resistance genes for 49 antibiotic-organism combinations, including gentamicin, tobramycin, ciprofloxacin, levofloxacin, trimethoprim-sulfamethoxazole, ertapenem, imipenem, cefazolin, cefepime, cefotaxime, ceftazidime, ceftriaxone, ampicillin, and aztreonam.”

The findings revealed that the test’s average positive predictive value for genotypic prediction of phenotypic resistance was 91% for E. coli, 93% for K. pneumoniae, 87% for P. mirabilis and 92% for P. aeruginosa, according to the study.

OpGen expects the first FDA clearances for the test this year, Walker noted.

“In high-risk patients needing urgent care and patients with complicated urinary tract infections, the faster determination of appropriate antibiotic therapy translates to improved health outcomes, lower hospital costs and better antibiotic stewardship,” Walker said. “OpGen’s technology is the application of precision medicine to infectious disease management, which has become more complex with the proliferation of antimicrobial resistance.”

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Advanced Detection Tool to Limit the Spread of Devastating Tree Pathogens

Seeking to prevent the introduction and spread of quarantine tree pathogens, the EU Horizon 2020-funded project HOMED (HOlistic Management of Emerging forest pests and Diseases) supports the development of an innovative tool for on-site detection of pathogens. The tool was developed by a team of scientists from the Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR) and the Department of Agrifood Production and Environmental Sciences (DISPAA), University of Florence and is described in an open access paper, published in the journal AMB Express.

Invasive alien species in trees and plants can have severe economic, ecological and sociological impact. Due to international trade, tourism and other human activities, non-native pathogens spread into new environments, causing a major threat to biodiversity, economy and human health. An effective framework for early warning and rapid response is a crucial element to intercept biological invasions of plant pathogens.

The HOMED-supported innovative tool works for on-site detection of quarantine pathogens, such as Xylella fastidiosa, Ceratocystis platani and Phytophthora ramorum. These are three of the most devastating pathogens of trees and ornamental plants accidentally introduced to Europe, which are causing epidemics with heavy consequences.

Requiring minimum equipment and easy to transport the new diagnostic tool can be also used by non-scientists to quickly and reliably check the health status of live trees and plants. Such user-friendly and compact tools greatly reduce the time usually necessary to take and analyse samples, thus allowing prompt reaction and increasing chances of confinement.

HOMED recognises the need for action and engages forest managers, biosecurity agencies, policy makers and environmental NGOs in the project with the goal to manage emerging forest diseases and thus preserve biodiversity.

Applying a holistic approach, HOMED improves strategies of risk assessment and management by targeting the successive phases of invasion, and developing mitigation methods for each phase - prevention, detection and diagnosis, surveillance, eradication and control tools.

The newly developed detection tool can be used to check the health status of live trees or tree parts, reducing time of analyses and in this way allowing a prompt reaction and potential control of environmental and economic losses. The tool allows a complete analysis in only 30 minutes time and can be applied as point-of-care diagnostics and represents a great advantage to preventing introductions and for applying control measures.

Author and HOMED partner Dr. Alberto Santini comments:

"The use of rapid, specific and sensitive point-of-care methods like the LAMP assays developed in this study could enable phytosanitary services to make immediate management decisions, helping in containing environmental and economic losses. The application of such a portable diagnostic tool, requiring minimum equipment and a few, if any, specific scientific skills could be profitably used to check the health status of live trees or tree parts at the points of entry or in field, thus reducing time of analyses and allowing a prompt reaction."

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binx health Receives CE Mark For Rapid Chlamydia And Gonorrhea Test For Mass Markets

binx health™, the pioneer in anywhere care today announced that its proprietary Chlamydia and Gonorrhea (CT/NG) test has received European CE Mark approval. The new multiplex test works with the binx io platform, delivering a laboratory-quality result in about 30 minutes, creating a new model for "test-and-treat" care for two of the most prevalent Sexually Transmitted Infections (STIs) globally.

More than one million STIs are acquired worldwide every day with chlamydia and gonorrhea representing two of the most prevalent1.  Recent data released by the European Centre for Disease Prevention and Control (ECDC) notes a striking rise in the cases of gonorrhea – up 17% between 2016 and 2017 across the reporting EU/EEA countries, with some countries noting year-over-year spikes of more than 40%2.  Similarly, in the United States, infection rates have risen an astonishing 67% since 2013, with rates continuing to be highest in the 20-24 year-old age group.3

Today, the majority of CT/NG tests are sent to central laboratories, meaning there can be up to a seven day waiting period from first visit to treatment, during which time patients may continue to spread infection, with up to 40% of positive patients never returning for treatment4. binx is positioned to radically change this paradigm, enabling patients and consumers to leave a retail pharmacy, primary care office, clinic, urgent care facility and any other brick-and-mortar location, with treatment in hand in a single visit (while the binx point-of-care platform is not yet available in the United States, the company recently completed its multi-center clinical study for submission to FDA for 510(k) clearance). The binx io platform is designed for ease-of-use, enabling non-laboratory-trained users to run a test with the same accuracy as a central lab test, making it ideal for use in a variety of approved medical and consumer settings.

"Achieving a 30-minute diagnostic turnaround time for two of the most common STIs is a major public health breakthrough," explains Howard Heller, MD MPH, binx health's Chief Medical Officer. "Rapid results equal rapid, appropriate treatment.  This new panel enables a path to not just same day, but same visit testing and treatment, eliminating an immense barrier to care and cure."

"Modern healthcare delivery requires tools that meet consumers' needs where they live, work and shop," said Jeffrey Luber, binx health's Chief Executive Officer. "Doing this effectively means delivering fast, accurate, cost-effective solutions at brick-and-mortar locations, as well as physician-ordered, medical guideline-based testing online, that work to bring consumers from 'worry to well' in the locations most convenient for them. Through scientific rigor, guideline-based testing, and an 'anywhere' mindset that puts the consumer at the center of all we do, today we embark on a path to deliver omnichannel, high quality care in family health, starting with young people and STIs as our launching point."

About binx health 

binx health™  is the pioneer in anywhere care for consumers where they live, work and shop. We do this with (1) proprietary, rapid desktop PCR instruments for retail pharmacy, urgent care, primary care, health centers, and any other brick-and-mortar location where onsite, on demand testing and rapid results on par with central lab testing, are paramount, and (2) mobile solutions for at-home, physician-ordered testing that aim to bring consumers from "worry-to-well" in the palm of their hands. Starting with sexually-transmitted infections, we aim to reverse trends of rising infections among young people, and move into broader family health through the expansion of our point-of-care and at home offerings. binx health's investors include financial investors, Johnson & Johnson Innovation, Novartis Venture Fund, LSP Venture Capital, BB Biotech, RMI Investments and Technology Venture Partners and strategic investors, Consort Medical (whose wholly owned subsidiary Bespak are the makers of the company's low-cost, proprietary multi-plex cartridges) and China-based Wondfo Biotech.

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RIT Professor Develops Device to Better Detect Ebola Virus

A faculty-researcher at Rochester Institute of Technology has developed a prototype micro device with bio-sensors that can detect the deadly Ebola virus. With this type of device, those infected can be treated earlier, and the early detection process can potentially decrease the spread of infections.

Ke Du, a faculty-researcher in RIT’s Kate Gleason College of Engineering, developed a microfluidic device that utilizes CRISPR gene-editing technology to monitor and detect the nucleic acid markers that indicate Ebola virus. The virus is highly contagious and there is limited treatment once an individual has been diagnosed, he said. There are several prominent strains of Ebola, and his research team has focused on the EBOV strain, which has a high mortality rate.

“If an individual travels from one infected community to another, they can easily spread the epidemic. That is why before any symptoms of Ebola, such as cough or fever present, individuals can take a blood test before being allowed to travel,” said Du, an assistant professor of mechanical engineering. He leads a multidisciplinary team of engineers and biochemists developing a rapid point-of care system and biochemistry array for in-field pathogen diagnosis. According to early results, the team has found that the Ebola RNA in test environments can be detected within five minutes by combining automated sample processing, fluorescence sensing and a unique CRISPR-Cas13a assay originated from a bacterial adaptive immune system.

The microfluidic device is an automated and small chip with a highly sensitive fluorescence sensing unit embedded into the device. Physicians take patient samples and add them into the device where Ebola RNA can be seen by activating the CRISPR mechanism. Du is also developing a device that could detect multiple virus strains from Ebola to influenza and zika, for example.

Du’s research was published in the April 2019 issue of ACS Sensors. The article “Rapid and Fully Microfluidic Ebola Virus Detection with CRISPR-Cas13a,” features an international and multidisciplinary team assessing the use of CRISPR technology—gene editing technology—to improve virus detection. The group members are from University of California, Berkeley; Tsinghua Berkeley Shenzhen Institute (China); Dong-A University (Korea); Texas Biomedical Research Institute; and Boston University.

“For this work, we are trying to develop a low-cost device that is easy to use especially for medical personnel working in developing countries or areas where there are outbreaks. They’d be able to bring hundreds of these devices with them for testing, not just one virus or bacteria at one time, but many different kinds,” he explained.

Researchers have tried for the past 40 years to develop an effective Ebola vaccine. Early detection remains an important strategy for controlling outbreaks, the most recent in the Congo, where more than 1,000 individuals have died, according to the Centers for Disease Control.

“If you look at this like influenza, and people don’t look at it as a virus which also can kill people each year. Some strains may not be as deadly as Ebola, but we know that infectious diseases, regardless of the type, are problems that can threaten the public,” Du said. “I grew up in China and experienced the 2002-2004 SARS outbreak. I have seen many people lose their relatives and friends because of infectious diseases. If we can have early detection systems to help screen for all types of diseases and patterns, this can be very useful because it can provide information to medical doctors and microbiologists to help develop the vaccines, and early detection and identification can control and even prevent outbreaks.”

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New 30-Minute Microarray Blood Test to Reduce Sepsis Deaths

A new detector using photonics to identify E. coli bacteria from a tiny drop of blood in less than 30 minutes could help reduce the mortality rate from sepsis by more than 70%.

Programmed to detect proteins and E. coli, one of the deadly bacteria that can cause the human body to go into septic shock, the detector uses light to look for specific biomarkers (the tell-tale signs or indicators of a disease) that are as small as a few nanometers in size, or 1/1,000,000th of the thickness of a single human hair.

The rapid microarray detector looks at a small blood sample taken from a thumb or forefinger. The patient’s blood sample is then separated in a centrifuge so that a clinician can examine the plasma, the part of the blood sample where all the proteins are contained.

The sepsis detector uses photonic technology to make a clear and accurate diagnosis. The plasma sample flows over a microarray, a collection of tiny spots containing specific antibodies on a nanostructured gold slide. Two light beams are then shone through the full microarray, with one of them passing through the sample, while the other one goes through the clear part of the slide, acting as a reference. The beams passing through the biomarker and the clear regions on the slide are then checked for any changes in intensity.

“Depending on the amount and type of biomarker attached to each antibody, we obtain a unique image, a signature pattern if you like,” said Roland Terborg, project coordinator. “The image patterns tell us what is present in the plasma sample, which we then record with a CMOS sensor, the same technology used in a digital camera that converts light into electrons.”

Preclinical trials have already begun at the Vall d'Hebron University Hospital where the device has been in operation since 2018. Clinical trials are expected to take place at the end of 2019.

Current techniques for detecting sepsis, a condition that kills more than 20,000 people per day worldwide, can take hours or even days to produce the results and diagnosis. While bacteria need to reproduce in large quantities before an accurate determination can be made, a patient can often be waiting more than a day before a course of treatment can be determined. If caught early enough, patients can be treated for sepsis with simple antibiotics.

The detector could also possibly be extended to perform other types of disease screenings or multiple simultaneous diagnoses, especially those requiring a rapid detection of large numbers of biochemical targets (more than one million) on a single microarray.

Developed by the RAIS project (or scalable point-of-care and label-free microarray platform for rapid detection of sepsis), the project was coordinated by ICFO in Barcelona and is a success story for the Photonics Public Private Partnership.

The RAIS consortium received a grant of €2,988,572 ($3,333,528) from the EU via the H2020 program and was made up of partners from Spain, Switzerland, Germany, Belgium, the U.K., and Italy. RAIS was coordinated by ICFO, and oversaw the collaboration of the Ecole Polytechnique Federale de Lausanne, iXscient, Diesse, the Catalan Institute of Nanoscience and Nanotechnology, microTEC, Trinean, Hospital Vall d’Hebron, and Thermo Fischer Brahms.

Additional information may be found on the RAIS Project website.

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3M Salmonella Molecular Test Validated for Animal Feed and Pet Food

3M Food Safety announced that the 3M Molecular Detection Assay 2 – Salmonella has earned matrix extensions from AFNOR Certification for its NF VALIDATION.

In February 2017, the test was first certified by AFNOR (Certificate reference number: 3M 01/16-11/16) for detection of Salmonella spp. in all human foods as well as in samples from the food processing environment, excluding primary production samples. The pathogen test’s validation now encompasses samples taken from primary production as well as from animal feed and pet food.

The Salmonella pathogen test kit is part of the 3M Molecular Detection System platform, award-winning technology that combines isothermal DNA amplification and bioluminescence detection to produce rapid, accurate results with fewer steps, reducing costs and technician time. Enhanced with a proprietary 3M nanotechnology, the test gives processors a streamlined workflow that enables same-day or next-day pathogen test results and faster, more confident food safety decisions. Comparative research shows that the 3M Molecular Detection Assay 2 – Salmonella will process a batch of 96 samples 1.7 times faster than the closest competitive technology. Last year, the 3M Molecular Detection System became a primary method of the USDA Food Safety and Inspection Service for the detection of both Salmonella and Listeria.

“We are committed to providing a wide range of customers with pathogen tests that have been rigorously and relevantly validated,” said Lisa Monteroso, 3M Food Safety regulatory affairs specialist. “Salmonella is an organism that can persist in a variety of foods – including those that are low-moisture – so we are pleased to share additional scientific support for our assay’s performance and use in these important categories.”

The NF VALIDATION certification process begins with a scientific comparison of testing methods; a single, expert lab thoroughly tests the technology’s effectiveness versus the NF EN ISO 6579 reference method. Subsequent to that, multiple laboratories coordinate to compare the efficacy of both the new test method and reference method under reproducible and repeatable conditions.

3M Food Safety is a leader of innovative solutions that help the food and beverage industries optimize the quality and safety of their products to enable consumer protection. It provides solutions that mitigate risk, improve operational efficiencies and impact bottom lines.

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Chembio Diagnostics Announces Approval of Zika, Dengue and Chikungunya Point-of-Care Multiplex Test by Brazil’s Health Regulatory Agency

Chembio Diagnostics, Inc., a leading point-of-care diagnostics company focused on infectious diseases, today announced the approval of its DPP Zika/Dengue/Chikungunya System by Agência Nacional de Vigilância Sanitária (ANVISA), Brazil’s health regulatory agency, in collaboration with Bio-Manguinhos/Fiocruz.

Chembio’s DPP Zika/Dengue/Chikungunya multiplex test allows simultaneous and discrete detection of antibodies for both active (IgM) and prior exposure (IgG) to the Zika, dengue, and chikungunya viruses, which is important for both treatment and surveillance. The test is performed using a small (10µL) drop of fingertip blood and provides quantitative results in approximately 15 minutes when used with the company’s handheld DPP Micro Reader. An evaluation of the multiplex test by Brazil’s Instituto Nacional de Controle de Qualidade em Saúde (INCQS) demonstrated outstanding analytical performance using patient samples in this highly endemic region, with all six results yielding sensitivity between 95.2% - 100% and specificity between 96.6% - 100%.

“Brazil faces a complex epidemiological scenario, characterized by simultaneous circulation of three arboviruses - Zika, dengue, and chikungunya - all transmitted by the Aedes aegypti mosquito,” stated John Sperzel, Chembio’s Chief Executive Officer. “Co-circulation hampers clinical differential diagnoses, as those arboviruses share common signs and symptoms. This ANVISA approval paves the way for commercialization in Brazil, and we believe our multiplex test will be an important tool in combating these serious diseases.”

This ANVISA approval advances Chembio’s previously announced commercial agreement with Bio-Manguinhos, a Brazilian government agency and supplier of diagnostic products to Brazil’s Ministry of Health, to develop and supply high-quality, rapid diagnostic tests for Zika, dengue, and chikungunya using Chembio’s DPP technology platform.

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GenePOC Obtains CE Marked for its GenePOC™ Carba Test

GenePOC Inc., member of the Debiopharm Group, announces its third test to be CE marked, the GenePOC Carba assay, to be used with the revogene™ device.

About carbapenem-producing organisms (CPO)

CPO are considered a serious global public health threat and are associated with significant morbidity, mortality and hospital costs.[1] In 2015, 13 of the 38 countries of the European Union and European Economic Area (EU-EEA) reported interregional spread or an endemic situation.2 High mortality rates, ranging from 30% to 75%, have been reported for patients with severe CPO infections.[3]

About the GenePOC Carba assay

The GenePOC Carba assay is a qualitative, in vitro diagnostic test designed for the detection and differentiation of the blaKPC, blaNDM, blaVIM, blaOXA-48-like, and blaIMP gene sequences associated with carbapenem-non-susceptibility.  The assay can provide results from one up to eight samples in approximately 70 minutes using characterized carbapenem-non-susceptible isolated colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa.

"The fight against carbapenemase-producing Enterobacteriaceae (CPE), a rising threat in healthcare facilities, requires rapid and efficient detection and differentiation," mentioned Professor Thierry Naas, MD, from the bacteriology services at Hôpital de Bicêtre, France.

It is important to note that CPO are adapted to spread in healthcare settings as well as in the community. To prevent transmission from CPO-positive patients, hospitals should consider enhanced infection control measures such as contact precautions, isolation and dedicated nurses for patients who are confirmed CPO-positive.[2]

"We are proud to receive the CE marked for our GenePOC Carba assay," said Patrice Allibert, PhD, CEO of GenePOC. "A test which offers rapid and accurate results will contribute to the identification of colonized patients, therefore limit the spread of these organisms in healthcare settings and save on hospital costs. Our GenePOC Carba assay also demonstrates the power of our technology to be compatible with panel detection," concluded Patrice Allibert.

About revogene

The revogene is an automated and stand-alone device, enabling testing of single-use proprietary microfluidic cartridges with fluorescence-based real-time polymerase chain reaction technology to deliver an accurate diagnosis.

About GenePOC

GenePOC, a member of the Debiopharm group, specializes in the development of diagnostic devices which enable the prevention and detection of infectious diseases.

References

1. Borer A et al. Infect Control Hosp Epidemiol. 2009;30(10) :972-6
2. https://ecdc.europa.eu/sites/portal/files/media/en/publications/Publications/carbapenem-resistant-enterobacteriaceae-risk-assessment-april-2016.pdf 
3. Tischendorf J. et al. Am J Infect Control. Epub 2016 Feb 15.

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New Biological Detection System Predicted to Provide Faster Less Expensive Results for Veterinarians

Veterinarians and agricultural inspectors who seek to detect and contain the spread of animal diseases can now turn to a newer, faster and less expensive biological detection system.

Known as the Axiom Microbiome Array, or AMA, the new biological detection system is the most comprehensive microorganism detection platform built to date and the first high-throughput microarray.

The AMA is the commercialized successor to Lawrence Livermore National Laboratory’s earlier microarray, called the Lawrence Livermore Microbial Detection Array (LLMDA).

The LLMDA was licensed in 2016 to Waltham, Massachusetts-based Thermo Fisher Scientific and went on sale later that year as Applied Biosystems AMA.

In a yearlong evaluation, published earlier this month in the online scientific journal PLOS ONE, a team of researchers from LLNL, Kansas State University and Thermo Fisher Scientific concluded that the microbiome array “is an efficient tool to rapidly analyze large numbers of clinical and environmental samples for the presence of multiple viral and bacterial pathogens.”

“Two of the array’s advantages are that it’s faster and cheaper,” said Raymond “Bob” Rowland, professor of diagnostic medicine and pathobiology at Kansas State’s College of Veterinary Medicine. “I live in the world of veterinary medicine and cost is critical.

“If we were to do a test using the LLMDA, it would have cost us about $250 just for the reagents; now with the AMA we can do the same tests for $40.”

When LLNL biologists and computer scientists first unveiled the versatile LLMDA in 2010, it could analyze samples for nearly 3,000 bacteria and viruses, all within 24 hours. But it could only analyze four samples a day.

With the 96-well AMA, the new detection system can analyze 96 samples in three days. Additionally, each of the 96 wells contain about 1.4 million probes, so samples are analyzed by about 132 million probes.

“One of the most important conclusions of our study is that this is the first high-throughput microarray that has been developed,” said LLNL biologist Crystal Jaing, who heads the LLMDA/AMA efforts. “It increases the throughput by 10- to 20-fold and decreases the cost by five-fold.”

As LLNL and Thermo Fisher Scientific researchers moved to a higher throughput microarray, one of the team’s prime goals has been to maintain a high-resolution and sensitivity of detection.

In their PLOS ONE paper, researchers from the three institutions note that with the new array, they were able to detect both Shigella, a food-borne bacterial pathogen, and Aspergillus, a fungal pathogen, at 100 genome copies, as well as vaccinia virus DNA, a genetic relation to smallpox, at 1,000 genome copies.

In veterinary tests with pigs, the microbiome array detected two viruses – porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 – as well as other common viral and bacterial microbiome species.

During their evaluation, the researchers conducted tests with 14 veterinary samples and 30 environmental samples, finding that the AMA performed at a resolution similar to the LLMDA, according to Jaing.

Because of the microbiome array, veterinarians can now conduct multiple tests on a single animal in Rowland’s view.

“This is a technology that is meant for the veterinary technician to use. It’s the everything test for everything.

“We base our testing on pathology. When we see a change in the tissue of an organ, that’s where your pathogen can usually be found. If we see changes in multiple organs and multiple tissues, now we can sample them all,” Rowland said.

According to the PLOS ONE paper authors, “Rapid detection and characterization of bacterial and viral pathogens is important for clinical microbiological diagnostics, public health, veterinary diagnostics, drug and food safety, environmental monitoring and biodefense.”

The current version of the AMA has the capability to detect more than 12,000 microorganisms and can identify 6,901 bacteria, 4,770 viruses and a combined total of 842 archaea, fungi and protozoa.

The new arrays, which come in 24- and 96-well formats, can analyze a variety of different sample types and can be used for nutrigenomics, agrigenomics, plus animal research and modeling.

In addition to Rowland and Jaing, the paper’s lead author, other paper co-authors are LLNL biologists James Thissen and Nicholas Be, LLNL bioinformaticists Kevin McLoughlin and Shea Gardner (deceased), retired LLNL computer scientist Tom Slezak and Thermo Fisher Scientific biologists Paul Rack and Michael Shapero.

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