Method of Testing RNA Unreliable

(Ivanhoe Newswire) -- Virginia Tech researchers have discovered an inconvenient truth about a frequently used assay to detect and measure mosquito-borne viral RNA in infected cells and tissues. The method most labs are using is unreliable.

The CHIKV virus has sickened millions of people in India and Africa in the last five years, and the ONNV virus has also caused large outbreaks of human disease with cases numbering in the millions. In studying virus infections borne by the mosquito, the Virginia Tech researchers developed a novel assay that is able to detect and measure anti-genomic copies of the viral genome. This differs from traditional assays that simply measure viral nucleic acids associated with infection, regardless of origin.

"The application of real-time PCR to the detection and quantification of specific strands of viral RNA is becoming an increasingly important tool in the study of RNA viruses. As a result, multiple types of assays have been described and are in widespread use. However, no study has yet determined if the accuracy with which the different types of assays detect and quantify specific strands of viral RNA are equivalent,” co-author and Assistant Professor of Entomology Kevin M. Myles was quoted as saying. “It turns out they are not, and the most frequently used method is the most error prone."
 
"A less frequently used . . . assay turned out to be more accurate," Entomology Assistant Professor and co-author Zach N. Adelman was quoted as saying. "The fact that many labs have been using assays prone to error may have led to some wrong conclusions," Adelman continued. "Using the more accurate assays will lead to more accurate conclusions and better science."

SOURCE: PLoS ONE, October 14, 2009

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