📄 Development and evaluation of a multitarget real-time Taqman reverse transcription-PCR assay for detection of the severe acute respiratory syndrome-associated coronavirus and surveillance for an apparently related coronavirus found in masked palm civets

Severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) is the etiological agent of SARS. It is believed that SARS-CoV originates from wild animals. We have developed a multitarget real-time Taqman reverse transcription-PCR (RT-PCR) assay for the quantitative detection of SARS-CoV. The sequences of the Taqman probes with a minor groove binder and the corresponding primers were based on the sequences of the N gene, open reading frame (ORF) 3, and ORF 8. The overall linear range of this assay was from at least 10 1 to 106 copies per reaction, and the detection limit could reach less than 10 copies per reaction. The quantification results for SARS-CoV from cell culture correlated well with those of the RT-PCR by using any two of the three sets of primer and probe used in this assay. However, the results of quantification of SARS-CoV obtained by using a few available throat swab specimens from SARS patients and the N gene as the target were almost 10 times higher than those obtained by using ORF 3 and ORF 8. Using this assay, we also detected an apparently SARS-CoV-related coronavirus in the throat swab specimens from masked palm civets in the west part of Hubei Province, People's Republic of China. Copyright © 2005, American Society for Microbiology.

keywords 🔗 reading frame (222) 🔗 etiological agent (62) 🔗 reverse transcription (205) 🔗 palm civets (20) 🔗 respiratory syndrome (2004) 🔗 acute respiratory (1734) 🔗 cell culture (240) 🔗 open reading (215) 🔗 throat swab (43)

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