A one-step, real-time PCR assay for rapid detection of rhinovirus.

The Journal of Molecular Diagnostics : JMD
Duc H DoRobert M Wadowsky

Abstract

One-step, real-time PCR assays for rhinovirus have been developed for a limited number of PCR amplification platforms and chemistries, and some exhibit cross-reactivity with genetically similar enteroviruses. We developed a one-step, real-time PCR assay for rhinovirus by using a sequence detection system (Applied Biosystems; Foster City, CA). The primers were designed to amplify a 120-base target in the noncoding region of picornavirus RNA, and a TaqMan (Applied Biosystems) degenerate probe was designed for the specific detection of rhinovirus amplicons. The PCR assay had no cross-reactivity with a panel of 76 nontarget nucleic acids, which included RNAs from 43 enterovirus strains. Excellent lower limits of detection relative to viral culture were observed for the PCR assay by using 38 of 40 rhinovirus reference strains representing different serotypes, which could reproducibly detect rhinovirus serotype 2 in viral transport medium containing 10 to 10,000 TCID(50) (50% tissue culture infectious dose endpoint) units/ml of the virus. However, for rhinovirus serotypes 59 and 69, the PCR assay was less sensitive than culture. Testing of 48 clinical specimens from children with cold-like illnesses for rhinovirus by the PCR and cult...Continue Reading

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Citations

Jun 22, 2012·Journal of Clinical Microbiology·Manuel SchiblerCaroline Tapparel
Feb 1, 2018·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Kim Tien NgKok Keng Tee
May 15, 2010·Expert Review of Molecular Diagnostics·Jane Kuypers
Aug 29, 2012·The Pediatric Infectious Disease Journal·Manuel SteinerAngelika Berger
Jan 17, 2020·Journal of Medical Virology·Naru ZhangShibo Jiang
Dec 22, 2020·American Journal of Infection Control·Alessandro ZulliJordan Peccia
Apr 1, 2021·Emergent Materials·E Alperay TarimH Cumhur Tekin
May 27, 2021·Nature Communications·Ramesh YelagandulaUlrich Elling

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