A repeat partnership

Rheonix, NYUCD to collaborate for rapid Zika virus diagnostic

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ITHACA, N.Y.—Rheonix Inc., a developer of fully automated molecular testing solutions, in collaboration with the New York University College of Dentistry (NYUCD), has received funding to develop a rapid diagnostic for Zika virus infection. The $656,414 award from the National Institute of Dental and Craniofacial Research (NIDCR) of the National Institutes of Health (NIH) will allow the Rheonix/NYUCD team to pursue the development of a fully automated screening and self-confirming assay that will simultaneously detect and confirm the presence of Zika virus in a small sample of saliva or blood. The proposed approach will build upon previous success in which the Rheonix/NYUCD team developed a dual assay for the simultaneous detection of HIV antibodies and viral RNA in a single specimen.
“In partnership with NYUCD, we were able develop a system that can detect both HIV antibodies and the actual viral RNA in a single specimen of saliva or blood, even in the earliest stages of the disease, to quickly confirm whether or not a patient is infected. We believe this system will significantly improve HIV testing by detecting early, acute HIV infection and addressing the well-known ‘seroconversion window’ when antibodies are not yet detectable, therefore eliminating the need for multiple patient visits to healthcare providers,” Dr. Greg Galvin, CEO and chairman of Rheonix, tells DDNews.
According to Galvin, the collaboration between Rheonix and NYUCD began five years ago. Dr. Daniel Malamud, who is professor of basic sciences and director of the HIV/AIDS Research Program at NYUCD, was the principal investigator on an NIH-funded grant that aimed to develop a rapid point-of-care test for HIV. Malamud and his team developed a novel assay, and then reached out to Rheonix to work together and automate the assay, Galvin says. This effort led to the Rheonix and NYUCD teams being awarded a Small Business Innovation Research Phase I/II Fast-track grant from the NIDCR to further simplify the original assay and to develop the microfluidic device and the instrument required to automate the process. The success of this technology has led the teams to collaborate again and pursue a rapid Zika virus diagnostic.
“We have had a longstanding and very productive collaborative relationship with Dr. Dan Malamud’s laboratory at NYUCD, and it has been through those efforts that we successfully developed the dual assay for anti-HIV antibodies and viral RNA,” commented Dr. Richard Montagna, senior vice president for scientific and clinical affairs at Rheonix and the principal investigator on the grant. “It seemed to be a logical extension of those efforts to attempt the same approach for the Zika virus.”
The assay for the Zika virus diagnostic will be performed on the Rheonix Chemistry and Reagent Device (Rheonix CARD) cartridge interfaced with the Encompass Optimum automated workstation. Once a raw sample is placed on the Rheonix CARD cartridge, the automated workstation runs with no user intervention through the process of sample extraction, purification, amplification and detection. This eliminates the need for multiple pieces of existing equipment, helping to make the testing process quicker, more efficient, less expensive and less likely to result in human error.
“As we continue to demonstrate the utility of our novel microfluidic-based technology, we remain committed to deploying the technology to address global health needs,” said Galvin. “Addressing the need to test for Zika virus certainly fits the bill, given the elevated health alerts issued throughout the world.”
In February 2016, the World Health Organization declared Zika virus a public health emergency of international concern. According to the Centers for Disease Control and Prevention, Zika virus disease is caused by Zika viruses that spread to people primarily through the bite of an infected Aedes aegypti or Aedes albopictus mosquito. Transmission from mother to child, through sexual contact and through blood transfusion have also been reported. According to the NIH, there is a tentative link between Zika virus infections in pregnant women and the occurrence of microcephaly in their newborn babies. In addition, a possible connection exists between Zika infection and Guillain-Barré syndrome, a condition in which the immune system attacks parts of the peripheral nervous system.
“The Zika virus appears to disappear from blood in six to 10 days, but is still detectable in saliva and urine,” explained Malamud. “Anti-Zika antibodies can be detected several days after infection. A combined RNA and antibody test will enable detection of both early and late Zika virus infections.”
According to Galvin, the teams plan to split the sample on the disposable Rheonix CARD cartridge using Rheonix’s microfluidic device. One portion of the sample will flow through a pathway to detect antibodies in a manner similar to an enzyme-linked immunosorbent assay. The other portion will be processed for molecular amplification using loop-mediated isothermal amplification (LAMP) technology with steps including viral lysis and extraction and purification of viral RNA. Finally, the sample will go through amplification of Zika-specific sequences by LAMP and detection.
“The development efforts proposed to the NIDCR were initiated within the past few weeks, and we hope to have a working prototype within the next few months,” says Galvin.

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