[Development of methods for detection of H5N1 from human clinical specimens].
BACKGROUND
To establish a method for H5N1 RNA detection and laboratory diagnosis of suspected human avian influenza (H5N1) virus infected cases.


METHODS
The typing and sub-typing primers were designed according to M and H5 and N1 gene respectively, and the RT-PCR and real-time PCR were developed using these primers.


RESULTS
The RT-PCR and real-time PCR could be used for H5N1 detection specifically, and there was no cross reaction with other influenza subtypes such as H1 and H3. The sensitivity for RT-PCR and real-time PCR was 1 TCID50 and 0.01 TCID50 respectively. Thirteen laboratory confirmed human H5N1 cases were detected from 42 suspected cases by using these methods.


CONCLUSIONS
The established RT-PCR and real-time PCR methods can be used for laboratory detection of suspected human H5N1 cases.
Wen
Le-ying
LY
Chinese Influenza Center, Institute for Viral Disease Control and Prevention, State Key Laboratory for Infectious Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Xu
Hong
H
Lan
Yu
Y
Zhao
Xiang
X
Zhang
Xiao-guang
XG
Wang
Da-yan
DY
Yao
Li-hong
LH
Dong
Jie
J
Zhang
Jia-huai
JH
Guo
Yuan-ji
YJ
Shu
Yue-long
YL
D000818:Animals; D002642:Chick Embryo; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics
D017931:DNA Primers; D014764:Viral Proteins