Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5

HUANG Bing; MA Xiu-li; WANG Li-li; LI Yu-feng; LIU Yu-shan; CHEN Fu-uan

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October 2005

HUANG Bing, MA Xiu-li, WANG Li-li, LI Yu-feng, LIU Yu-shan and CHEN Fu-uan. Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5[J]. Virologica Sinica, 2005, 20(5): 498-502.

Citation: HUANG Bing, MA Xiu-li, WANG Li-li, LI Yu-feng, LIU Yu-shan, CHEN Fu-uan. Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5 .VIROLOGICA SINICA, 2005, 20(5) : 498-502.

H_9与H_5亚型禽流感病毒血凝素基因的快速鉴别

黄兵 ^1,2 ,
马秀丽 ² ,
王莉莉 ² ,
李玉峰 ² ,
刘玉山 ² ,
陈溥言 ^1*

1.
1.山东省农业科学院家禽研究所山东济南 2.江苏南京南京农业大学农业部动物疫病诊断与免疫重点开放实验室

摘要

建立一步法RT-PCR检测方法,对禽流感病毒(Avian influenza virus,AIV)的血凝素(Hemagglutinin,HA)分型进行了研究。参照AIV的HA基因序列设计1对引物,对H9和H5亚型AIV进行了扩增,产物大小分别为579bp和177bp。经测试,该引物不与新城疫病毒等鸡的其它传染性病原及鸡肌肉组织的核酸发生交叉反应。敏感性分析发现,从50pg的AIV总RNA中亦能扩增到目的条带。结果表明,此次利用1对引物建立的一步法RT-PCR方法简便适用,可以在一次反应中同时将H9和H5亚型AIV进行快速检测和分型。另外,两个亚型的扩增产物均包含了HA裂解位点在内的基因序列,可通过测序推导氨基酸顺序以预测H5或H9亚型禽流感病毒的潜在毒力。
- 禽流感病毒
- , 血凝素基因
- , 一步法RT-PCR
- , 分型

Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5

HUANG Bing ^1,2 ,
MA Xiu-li ² ,
WANG Li-li ² ,
LI Yu-feng ² ,
LIU Yu-shan ² ,
CHEN Fu-uan ^1*

Abstract

A method of one-step reverse transcription polymerase chain reaction was developed to subtype hemagglutinin(HA) gene of Avian influenza virus(AIV).One pair of primers were designed on the basis of HA gene of AIV,allowing simultaneous detection of AIV subtype H_9 and H_5,and the specific amplicons were 579bp and 177bp in size respectively.The primers were specific for AIV(H_9 and H_5 subtype) with no cross-reaction to RNA from the chicken muscle and other chicken pathogens such as Newcastle disease virus etc.The sensitivity of this assay was about 50pg of total RNA of AIV.Our data showed that this method was simple and feasible for rapid identification and subtyping of AIV subtype H_9 and H_5 at a single reaction.In addition,both diagnostic amplicons for AIV subtype H_9 and H_5 contained the cleavage site sequence of HA gene,from which the amino acid sequence might be deduced to predict the potential virulence of AIV.
- Avian influenza virus（AIV）
- , Hemagglutinin gene
- , One-step
- , RT-PCR

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Rapid Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H9 and H5

Abstract: A method of one-step reverse transcription polymerase chain reaction was developed to subtype hemagglutinin(HA) gene of Avian influenza virus(AIV).One pair of primers were designed on the basis of HA gene of AIV,allowing simultaneous detection of AIV subtype H_9 and H_5,and the specific amplicons were 579bp and 177bp in size respectively.The primers were specific for AIV(H_9 and H_5 subtype) with no cross-reaction to RNA from the chicken muscle and other chicken pathogens such as Newcastle disease virus etc.The sensitivity of this assay was about 50pg of total RNA of AIV.Our data showed that this method was simple and feasible for rapid identification and subtyping of AIV subtype H_9 and H_5 at a single reaction.In addition,both diagnostic amplicons for AIV subtype H_9 and H_5 contained the cleavage site sequence of HA gene,from which the amino acid sequence might be deduced to predict the potential virulence of AIV.