Performance of Single-Step Gel-Based Reverse Transcription-PCR (RT-PCR) Assays Equivalent to That of Real-Time RT-PCR Assays for Detection of the Severe Acute Respiratory Syndrome-Associated Coronavirus

Simple gel-based one-step reverse transcription-PCR (RT-PCR) assays, used to investigate patients during the 2003 severe acute respiratory syndrome (SARS) outbreak in Singapore, were found to be as sensitive as commercial and in-house real-time RT-PCR assays. The detection limit was approximately 1...

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Bibliographic Details
Published in:Journal of Clinical Microbiology 2005-08, Vol.43 (8), p.4262-4265
Main Authors: Inoue, Masafumi, Barkham, Timothy, Keong, Lee Kok, Gee, Lim Seng, Wanjin, Hong
Format: Article
Language:English
Subjects:
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Humans
Infectious diseases
Medical sciences
Microbiology
Miscellaneous
Reverse Transcriptase Polymerase Chain Reaction - methods
SARS Virus - genetics
SARS Virus - isolation & purification
Sensitivity and Specificity
Virology
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Summary:Simple gel-based one-step reverse transcription-PCR (RT-PCR) assays, used to investigate patients during the 2003 severe acute respiratory syndrome (SARS) outbreak in Singapore, were found to be as sensitive as commercial and in-house real-time RT-PCR assays. The detection limit was approximately 1 genome equivalent (GE) per 5 [micro]l PCR mixture. One PFU of SARS coronavirus was estimated to be 258 ± 46 GE.
ISSN:0095-1137
1098-660X
DOI:10.1128/JCM.43.8.4262-4265.2005