Establishment of a fluorescent polymerase chain reaction method for the detection of the SARS-associated coronavirus and its clinical application.
OBJECTIVE
To establish a fluorescent polymerase chain reaction (F-PCR) method for detecting the coronavirus related to severe acute respiratory syndrome (SARS) and to evaluate its value for clinical application.


METHODS
The primers and the fluorescence-labeled probe were designed and synthesized according to the published sequence of the SARS-associated coronavirus genes. A F-PCR diagnosis kit for detecting the coronavirus was developed, and 115 clinical nasopharyngeal gargling liquid samples were tested.


RESULTS
The sequence of PCR amplified products completely matched the related sequence of the SARS-associated coronavirus genome. Forty-nine out of 67 samples from identified SARS patients and 8 of 18 samples from persons having close contact with SARS patients showed positive results. All 30 samples from healthy controls were negative.


CONCLUSIONS
The F-PCR method established may be a rapid, accurate and efficient way for screening and for the early diagnosis of SARS patients.
Wu
Xinwei
X
Guangzhou Centre for Disease Control and Prevention, Guangzhou 510080, China. tom.wu@tom.com
Cheng
Gang
G
Di
Biao
B
Yin
Aihua
A
He
Yunshao
Y
Wang
Ming
M
Zhou
Xinyu
X
He
Lijuan
L
Luo
Kai
K
Du
Lin
L
D005453:Fluorescence; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D045169:Severe Acute Respiratory Syndrome / Q000821:virology