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. 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.
Dewhurst-Maridor
G
G
Division of Clinical Pathology, CMU, Michel-Servet 1, Ch-1211 Geneva 4, Switzerland.
Simonet
V
V
Bornand
J E
JE
Nicod
L P
LP
Pache
J C
JC
D052160:Benzothiazoles; D003959:Diamines; D006801:Humans; D016867:Immunocompromised Host; D016377:Organ Transplantation; D009930:Organic Chemicals; D011804:Quinolines; D015203:Reproducibility of Results; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I