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

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. Althoug...

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Bibliographic Details
Published in:Journal of virological methods 2004-09, Vol.120 (1), p.41-49
Main Authors: Dewhurst-Maridor, G, Simonet, V, Bornand, J.E, Nicod, L.P, Pache, J.C
Format: Article
Language:English
Subjects:
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Humans
Immunocompromised Host
Immunocompromised patients
Microbiology
Organ Transplantation
Organic Chemicals
Paramyxoviridae
Quantitative RT–PCR
Reproducibility of Results
Respiratory syncytial virus
Respiratory syncytial virus (RSV)
Respiratory Syncytial Virus Infections - diagnosis
Respiratory Syncytial Virus Infections - virology
Respiratory Syncytial Viruses - genetics
Respiratory Syncytial Viruses - isolation & purification
Reverse Transcriptase Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction - standards
Sensitivity and Specificity
Staining and Labeling
SYBR Green
TaqMan
Techniques used in virology
Virology
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Summary: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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2004.03.017