Rapid and sensitive detection of avian influenza virus subtype H7 using NASBA

被引:38
作者
Collins, RA
Ko, LS
Fung, KY
Chan, KY
Xing, J
Lau, LT
Yu, ACH
机构
[1] Hong Kong DNA Chips Ltd, Kowloon, Hong Kong, Peoples R China
[2] Peking Univ, Neurosci Res Inst, Beijing 100083, Peoples R China
[3] Peking Univ, Dept Neurobiol, Beijing 100083, Peoples R China
关键词
NASBA; avian influenza; H7; haemagglutinin; electrochemiluminescence;
D O I
10.1016/S0006-291X(02)02896-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) is an isothermal technique allowing rapid amplification and detection of specific regions of nucleic acid from a diverse range of sources. It is especially suitable for amplifying RNA. A NASBA/ECL technique has been developed allowing the detection of RNA from avian influenza virus subtype H7 derived from allantoic fluid harvested from inoculated chick embryos and from cell cultures. Degenerate amplification primers and amplicon capture probes were designed enabling the detection of low and highly pathogenic avian influenza of the H7 subtype from the Eurasian and North American lineages and the Australian sub-lineage. The NASBA/ECL technique is specific for subtype H7 and does not cross-react with other influenza subtypes or with viruses containing haemagglutinin-like genes. The assay is 10- to 100-fold more sensitive than a commercially available antigen capture immunoassay system. The NASBA/ECL assay could be used in high throughput poultry screening programmes. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:507 / 515
页数:9
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