Key result
A two-tube multiplex RT-PCR assay for detecting sixteen respiratory viruses yielded no false positives and six false negatives among 247 evaluated specimens.
Why the study?
Does a two-tube multiplex RT-PCR assay accurately detect sixteen common respiratory viruses compared to established assays in clinical specimens?
Does a two-tube multiplex RT-PCR assay accurately detect sixteen common respiratory viruses compared to established assays in clinical specimens?
The developed two-tube multiplex RT-PCR assay is a sensitive and specific tool with great potential for routine surveillance of respiratory virus infections.
Supports surveillance use with high specificity; extends multiplex RT-PCR options but leaves open larger validation studies.
There is a need for the development of a rapid and sensitive diagnosis of respiratory viral pathogens. With an intended application in provincial Centers for Diseases Control and Prevention, in this study, we present a two-tube multiplex RT-PCR assay (two-tube assay) using automatic electrophoresis to simultaneously detect sixteen common respiratory viruses. The specificity and the sensitivity of the assay were tested. The assay could detect 20-200 copies per reaction when each viral type was assayed individually, 2000 copies with 9 premixed viral targets in the multiplexed assay in tube 1, and 200 copies with 8 premixed templates in tube 2. A total of 247 specimens were used to evaluate the two-tube assay, and the results were compared with those obtained from the Luminex xTAG RVP Fast assay. The discordant results were confirmed by sequencing or by the Seeplex RV15 ACE detection kit. There were no false positives, but six false negatives occurred with the two-tube assay. In conclusion, the two-tube assay is demonstrated to have great potential for routine surveillance of respiratory virus infection in China.
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Li et al. (2013) studied Respiratory virus infection (n=247). Two-tube multiplex RT-PCR assay vs. Luminex xTAG RVP Fast assay was evaluated on Detection of respiratory viruses (false positives and false negatives). A two-tube multiplex RT-PCR assay for detecting sixteen respiratory viruses yielded no false positives and six false negatives among 247 evaluated specimens.
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