Detecting Highly Pathogenic Avian Influenza Virus H5N1 Subtype for Human Being by RT-Multiplex PCR
- VernacularTitle:逆转录多重PCR检测人感染高致病禽流感H5N1亚型
- Author:
Ding LIU
;
Zheng WANG
;
Yi ZHANG
;
Yuanchao LI
;
Ping CHEN
;
Fenglin L
- Publication Type:Journal Article
- Keywords:
Avian influenza virus;
Highly pathogenic avian influenza virus;
H5N1;
RT-M PCR
- From:
Chinese Journal of Nosocomiology
2009;0(13):-
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To establish and optimize the detection of highly pathogenic influenza virus H5N1 subtype by RT-Multiplex (RT-M) PCR method. METHODS The viral RNA was reversely transcripted with universal primer,and the cDNA was sequenced and analyzed. According to the cleavage site of H5 and the conservative sequence of N1,designed two pairs of specificity primer,RT-M PCR was developed by these primers. Then verified the sensitivity of method and its specificity by detecting comparing with NDV,IBV and IBDV. RESULTS The results of H5N1 gene sequence showed the high homology with the Anhui H5N1 strain [avian influenza A/Anhui/1/2005 (H5N1)] after comparing in the GenBank. Two fragments of 302 bp and 567 bp were amplified by RT-M PCR. The method could detect 0.500 pg virus RNA at least,and own high specificity. CONCLUSIONS This method can be used for highly pathogenic avian influenza.
