[Determination of SARS-CoV by simplified nested fluorescent RT-PCR].
OBJECTIVE
To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR.


METHODS
Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested.


RESULTS
The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160).


CONCLUSIONS
The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.
Li
Zhen-yong
ZY
Key Laboratory of Chem-biology, Department of Chemistry, Zhengzhou University, Zhengzhou 450052, China. zhenyongl@sabc.com.cn
Shao
Da-xiao
DX
Li
Zhi-tao
ZT
Feng
Yan-ming
YM
Jing
Jian-zhou
JZ
Wang
Qui-qi
QQ
Wang
Yun-long
YL
Qu
Ling-bo
LB
Zhao
Yu-fen
YF
D012634:Bodily Secretions / Q000821:virology; D017931:DNA Primers; D006801:Humans; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000097:blood / Q000175:diagnosis / Q000821:virology*
D017931:DNA Primers; D012367:RNA, Viral