Widely used virus assay shown unreliable when compared to other methods

Oct 21, 2009

In the course of doing research on the mosquito-borne pathogens chikungunya virus (CHIKV) and o' nyong-nyong virus (ONNV), Virginia Tech researchers have discovered an inconvenient truth about an assay, strand-specific quantitative real-time PCR (ssqPCR), increasingly being used to detect and measure replicating viral RNA in infected cells and tissues. The method most labs are using for ssqPCR is unreliable.

The research appears in the Wednesday, October 14, 2009, issue of , in the article, "Accurate Strand-Specific Quantification of Viral RNA," by Nicole E. Plaskon of Richmond Va., a Master of Science in life sciences candidate in the College of Agriculture and Life Sciences, and entomology Assistant Professors Zach N. Adelman and Kevin M. Myles, all with the Fralin Life Science Institute..

CHIKV has sickened millions of people in India and Africa in the last five years - 1.3 million in India alone. ONNV has also previously caused large outbreaks of human disease with cases numbering in the millions. In studying of the mosquito, the Virginia Tech researchers developed a novel assay that detects and measures anti-genomic copies of the . 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 ssqPCR 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 are equivalent. It turns out they are not, and the most frequently used method is the most error prone," said Myles.

"A less frequently used ssqPCR assay turned out to be more accurate," said Adelman.

"The fact that many labs have been using assays prone to error may have led to some wrong conclusions," Adelman said. "Using the more accurate assays will lead to more accurate conclusions and better science."

Although Myles and Adelman developed their assays for CHIK and ONNV, the results should help improve the design of ssqPCR assays for the study of other RNA viruses as well.

More information: The PLOS One (Public Library of Science) article is available on line at dx.plos.org/10.1371/journal.pone.0007468

Source: Virginia Tech (news : web)

Explore further: Premature aging: Scientists identify and correct defects in diseased cells

Related Stories

Researchers create first chikungunya animal model

Feb 19, 2008

Researchers have developed the first animal model of the infection caused by chikungunya virus (CHIKV), an emerging arbovirus associated with large-scale epidemics that hit the Indian Ocean (especially the French Island of ...

Recommended for you

Why you need one vaccine for measles and many for the flu

16 hours ago

While the influenza virus mutates constantly and requires a yearly shot that offers a certain percentage of protection, old reliable measles needs only a two-dose vaccine during childhood for lifelong immunity. ...

Scientists turn blood into neural cells

16 hours ago

Scientists at McMaster University have discovered how to make adult sensory neurons from human patients simply by having them roll up their sleeve and providing a blood sample.

How our gut changes across the life course

18 hours ago

Scientists and clinicians on the Norwich Research Park have carried out the first detailed study of how our intestinal tract changes as we age, and how this determines our overall health.

User comments : 0

Please sign in to add a comment. Registration is free, and takes less than a minute. Read more

Click here to reset your password.
Sign in to get notified via email when new comments are made.