Development of a quantitative TaqMan RT-PCR for respiratory syncytial virus

Journal of Virological Methods
G Dewhurst-MaridorJ C Pache

Abstract

Respiratory syncytial virus (RSV) is a ubiquitous RNA virus of the family Paramyxoviridae that may interfere with graft tolerance and with other interstitial lung diseases. The low viral titre observed in the immunodeficient transplanted patients requires a highly sensitive detection method. Although different tests already exist for the detection of RSV, reverse transcription-polymerase chain reaction (RT-PCR) has been shown to have the best sensitivity. In this study, a SYBR Green assay was established for the detection of RSV A and RSV B in a common screening test, and two quantitative TaqMan RT-PCRs were developed to quantify both RSV subgroups separately. Standard dilutions obtained from RSV cell infections were included in each test, and the assay was normalised using a housekeeping gene. RSV was found in 16% of the transplanted patients tested. The quantitative TaqMan assay is fast, reproducible, specific and very sensitive, and could facilitate considerably the detection of RSV virus. This would in-turn facilitate studies on the role of RSV in graft rejection.

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Citations

Jun 30, 2009·The Pediatric Infectious Disease Journal·Valerie WatersScott A Halperin
Jan 3, 2014·Advances in Virology·Swapnil Subhash BawageShree Ram Singh
Sep 21, 2012·Scandinavian Journal of Infectious Diseases·Rogier A S HoekMenno M van der Eerden
Oct 20, 2009·The Journal of Heart and Lung Transplantation : the Official Publication of the International Society for Heart Transplantation·Ilker UçkayChristian van Delden
Sep 7, 2013·Journal of Medical Virology·A B TedcastleG L Toms
Apr 6, 2016·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·F M MoeskerP L A Fraaij
Apr 26, 2005·Journal of Virological Methods·D DoniaA Pana'
May 29, 2012·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·R SlingerF Chan

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