The design and application of DNA chips for early detection of SARS-CoV from clinical samples.
BACKGROUND
SARS coronavirus has been identified as the cause of severe acute respiratory syndrome (SARS). Few tests allow confirmation or exclusion of SARS within the first few days of infection. A gene chip is a useful tool for the study of microbial infections mainly for its capability of performing multi-target analysis in a single test.


OBJECTIVE
Investigate the possibility of early detection of SARS virus from clinical samples using the gene chip-based method.


METHODS
We purified RNA from SARS-CoV obtained from routinely collected peripheral blood and sputum samples of 34 patients who had been identified as probable SARS patients by following the interim U.S. case definition. Four segments of the SARS-CoV were amplified using reverse transcription-nested PCR and the products examined using the 70-mer gene chips for SARS-CoV detection.


RESULTS
A blind-test of both peripheral blood and sputum specimens lead to the positive detection of SARS-CoV in 31 out of 34 patients. SARS-CoV was not found in peripheral blood or sputum specimens from three patients. Two of the 34 patients were only 3 days post-onset of symptoms and were subsequently confirmed to be SARS positive. Our results indicate that the gene chip-based molecular test is specific for SARS-CoV and allows early detection of patients with SARS with detection rate about 8% higher than the single PCR test when the sputum sample is available.
Zhou
Yi-Ming
YM
State Key Laboratory of Biomembrane and Membrane Biotechnology, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Yang
Ren-quan
RQ
Tao
Sheng-ce
SC
Li
Ze
Z
Zhang
Qiong
Q
Gao
Hua-fang
HF
Zhang
Zhi-wei
ZW
Du
Jian-yu
JY
Zhu
Pei-xuan
PX
Ren
Li-li
LL
Zhang
Liang
L
Wang
Dong
D
Guo
Lan
L
Wang
Yan-bin
YB
Guo
Yong
Y
Zhang
Yan
Y
Zhao
Chuan-zan
CZ
Wang
Can
C
Jiang
Di
D
Liu
Yan-hua
YH
Yang
Hua-wei
HW
Rong
Li
L
Zhao
Yan-jun
YJ
An
Shuang
S
Li
Zhe
Z
Fan
Xiao-dong
XD
Wang
Jian-wei
JW
Cheng
Yun
Y
Liu
Ou
O
Zheng
Zhong
Z
Zuo
Huan-cong
HC
Shan
Quan-zhong
QZ
Ruan
Li
L
Lü
Zhan-xiu
ZX
Hung
Tao
T
Cheng
Jing
J
D001769:Blood / Q000821:virology; D002681:China; D042241:Early Diagnosis; D006801:Humans; D025202:Molecular Diagnostic Techniques; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis* / Q000235:genetics; D013183:Sputum / Q000821:virology
D012367:RNA, Viral