NEW YORK, U.S.: Early detection of any disease or virus is critical. The 2015 Zika outbreak in Brazil, which led the World Health Organization to declare Zika and its link to birth defects a public health emergency, is a prime example of this. Now, researchers at the New York University College of Dentistry (NYU Dentistry) are developing a novel test for the Zika virus that uses saliva to identify diagnostic markers of the virus in a fraction of the time of current commercial tests.
Working with the research company Rheonix, the scientists built upon their earlier collaborative work into the detection of HIV, for which they developed a rapid saliva test that can recognize both viral RNA and antibodies. “When we developed the HIV test, we knew we could use the same model for any infectious disease. All we need to know is the nucleic acid sequence and an antigen to identify specific antibodies,” said study author Dr. Daniel Malamud, Professor of Basic Science at NYU Dentistry.
Testing for pathogens often involves two separate steps: one to detect a pathogen’s nucleic acids (RNA or DNA) and another to test for antibodies. Currently, the main way of testing for Zika is through real-time polymerase chain reaction tests, which can take around 3 hours—with specific antibody tests taking up to several weeks. Blood is usually used to test for the virus; however, the researchers believe that it may not be the best option, as the virus’s nucleic acids disappear in the blood a week or two after a person is infected, but can persist longer in saliva, semen and urine. In addition, antibodies can remain for months or years in those bodily fluids, which is why it is essential for diagnosis to also detect antibodies after infection.
Funded by the National Institutes of Health, the NYU Dentistry researchers are now developing a test that combines both nucleic acid and antibody assays using saliva, given that the Zika virus’s nucleic acids and antibodies persist in this fluid. A saliva test is also noninvasive, cost-effective and easier to collect than blood or urine. According to the researchers, the new test has the potential to produce results in a matter of minutes.
“The sooner you can identify a pathogen, the sooner steps can be taken to treat and isolate people. During an epidemic, you could test people before they get on a plane. The future of going through security at the airport may not be taking off your shoes, but instead spitting into a tube,” said Malamud.
For the test, the researchers altered the model to detect Zika nucleic acid sequences instead of those for HIV. They then used a portable isothermal amplification device—which could be used for point-of-care testing—to identify Zika RNA. With evidence showing that both the nucleic acid and antibody tests work, the researchers can combine them in the Rheonix CARD cartridge, a fully integrated microfluidic device, to process both diagnostic assays automatically and simultaneously.
The test is described in two studies, titled “Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva” and “Zika Virus Specific Diagnostic Epitope Discovery,” which were published in PLOS ONE and the Journal of Visualized Experiments on Feb. 5, 2018, and Dec. 12, 2017, respectively.
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