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| H5、H7和H9亚型禽流感病毒多重反转录聚合酶链式反应检测方法的建立 | |
| 引用本文： | 张文慧,王伟利,郑聪,刘明,钱爱东. H5、H7和H9亚型禽流感病毒多重反转录聚合酶链式反应检测方法的建立[J]. 中国兽医学报, 2010, 30(3) |
| 作者姓名： | 张文慧 王伟利 郑聪 刘明 钱爱东 |
| 作者单位： | 1. 吉林农业大学,吉林,长春,130118 2. 吉林出入境检验检疫局,吉林,长春,130062 3. 中国农业科学院,哈尔滨兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001 |
| 基金项目： | 吉林省科技厅资助重大项目(20065020) |
| 摘 要： | 参考GenBank登录的H5、H7和H9亚型禽流感病毒(AIV)的血凝素基因序列,利用DNAStar软件分析其同源性,利用Primer Premier5.0软件设计3对分别针对AIVH5、H7和H9亚型的特异性引物。3对引物所扩增的cDNA片段大小分别为427、228、830bp。结果显示,利用3对引物,通过对多重RT-PCR扩增条件的优化,建立了同时检测H5、H7和H9亚型AIV的多重RT-PCR技术。该方法对H5、H7和H9亚型AIV能同时扩增出3条大小分别为427、228、830bp的cDNA片段,与其他常见禽病病原不存在交叉反应。该方法对H5、H7和H9亚型AIVcDNA的最低检出量分别为10-4、10-2和10-4。结果表明,本试验建立了可同时检测鉴别H5、H7和H9亚型AIV的多重RT-PCR技术。 |
| 关 键 词： | 多重反转录聚合酶链式反应 禽流感病毒 H5亚型 H7亚型 H9亚型 |
Development of multiplex reverse transcription polymerase chain reaction for identification of H5,H7 and H9 subtypes of avian influenza virus |
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| ZHANG Wen-hui,WANG Wei-li,ZHENG Cong,LIU Ming,QIAN Ai-dong. Development of multiplex reverse transcription polymerase chain reaction for identification of H5,H7 and H9 subtypes of avian influenza virus[J]. Chinese Journal of Veterinary Science, 2010, 30(3) | |
| Authors: | ZHANG Wen-hui WANG Wei-li ZHENG Cong LIU Ming QIAN Ai-dong |
| Affiliation: | ZHANG Wen-hui1,WANG Wei-li2,ZHENG Cong1,LIU Ming3,QIAN Ai-dong1 (1.Jilin Agricultural University,Changchun 130118,China,2.Jilin Entry-Exit Inspection , Quarantine Bureau of People's Republic of China,Changchun 130062,3.National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute of CAAS,Harbin 150001,China) |
| Abstract: | H5,H7 and H9 subtypes avian influenza virus hemagglutinin gene sequences were searched in GenBank,and their homology were analyzed using DNAStar.The sets of specific oligonucleotide primers were designed.The results of this specific multiplex RT-PCR demonstrated that the specific cDNA bands of 427 bp,228 bp and 830 bp were observed simultaneously for RNA isolated from H5,H7 and H9 subtypes AIV.There was no cross-reaction with other avian pathogens.The minimal amount of detection was 10-4 cDNA of H5,10-2 cDN... |
| Keywords: | multiplex RT-PCR avian influenza virus H5 subtype H7 subtype H9 subtype |
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