Identification and subtyping of avian influenza viruses by reverse transcription-PCR
Identification and subtyping of avian influenza viruses by reverse transcription-PCR
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DOI：
10.1016/s0166-0934(01)00301-9
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发表时间：
2001-09-01
影响因子：
3.1
通讯作者：
Shieh, HK
中科院分区：
文献类型：
--
作者：
Lee, MS;Chang, PC;Shieh, HK
禽流感病毒有15种不同的血凝素（HA）亚型（H1-H15）。本文报道了一种用逆转录聚合酶链反应（RT-PCR）鉴定禽流感病毒HA亚型的方法。禽流感病毒通过RT-PCR使用一组对禽流感病毒的核蛋白（NP）基因具有特异性的引物进行鉴定。通过同时运行15个RT-PCR反应来确定禽流感病毒的HA亚型，每个RT-PCR反应使用一组对一种HA亚型具有特异性的引物。对于单一病毒株或分离株。15个RT-PCR反应中只有一个会产生预期大小的产物，因此确定了病毒的HA亚型。HA亚型的结果，然后通过PCR产物的序列分析确认。共80株禽流感病毒或分离株的亚型通过这种RT-PCR程序。RT-PCR检测结果与常规血清学方法检测结果的相关性为100%。我们开发的RT-PCR方法是快速和敏感的。可用于器官匀浆中禽流感病毒的鉴定和HA亚型分型。(C)2001 Elsevier Science B. V.保留所有权利。
Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1-H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate. only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure. and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive. and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates. (C) 2001 Elsevier Science B.V. All rights reserved.