[Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR].
OBJECTIVE
To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus.


METHODS
The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis.


RESULTS
Specific fragments were amplified from the sputum samples of SARS patients, which were confirmed by DNA cloning and sequencing to belong to SARS-associated coronavirus. The Result of Blast shows only the difference in one nucleic acid from the TOR2 strain of SARS-associated coronavirus.


CONCLUSIONS
Sequence analysis has confirmed the existence of SARS-associated coronavirus in the sputum samples of SARS patients, and nested RT-PCR is a quick, easy, and convenient way for the detection of the virus.
Wang
Yan
Y
Institute of Genetic Engineering, First Military Medical University, Guangzhou 510515, China.
Ma
Wen-li
WL
Song
Yan-bin
YB
Xiao
Wei-wei
WW
Zhang
Bao
B
Huang
Hai
H
Wang
Hong-min
HM
Ma
Xiao-dong
XD
Zheng
Wen-ling
WL
D001483:Base Sequence; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000302:isolation & purification
D012367:RNA, Viral