| 人源性甲型流感病毒(H1N1)分离株血凝素生物学特征分析 |
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| 引用本文： | 王华洪,袁丹,何太平. 人源性甲型流感病毒(H1N1)分离株血凝素生物学特征分析[J]. 实用预防医学, 2010, 17(12): 2386-2388 |
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| 作者姓名： | 王华洪 袁丹 何太平 |
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| 作者单位： | 1. 湖南省岳阳市疾病预防控制中心,湖南,岳阳,414000
2. 广东医学院 |
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| 摘 要： | 目的分离人甲型H1N1流感病毒,探讨血凝素基因的生物学特征。方法采集流感患者鼻咽分泌物,MDCK细胞进行病毒分离,血凝抑制实验测定病毒滴度,提取病毒总RNA,设计特异性引物,RT-PCR扩增HA基因ORF开放阅读框cDNA,克隆到pUCm-T载体,转化大肠杆菌XL1-Blue,经蓝白斑筛选阳性克隆,PCR鉴定后进行基因序列测定,通过生物信息学软件对HA的基因组结构和生物学特征进行分析。结果通过MDCK细胞培养成功分离出人甲型H1N1流感病毒,并于接种后24 h出现CPE,第二代病毒培养物滴度为256;成功扩增出产物大小为1 701 bp的H1N1流感病毒HA基因;构建了pUCm-T/HA载体并测序成功;BLAST结果显示与A/California/04/2009(H1N1)病毒株核苷酸与氨基酸同源性均为99%,所分离的毒株包含20种566个氨基酸。结论甲型H1N1流感病毒的分离和HA基因的测序,为后续HA蛋白的功能学研究奠定了坚实基础。
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| 关 键 词： | A型流感病毒 病毒分离 HA 生物学特征 |
Antigenic and Genetic Characteristics of Hemagglutinin Gene of H1N1 Influenza Viruses Circulating in Human |
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| WANG Hua-hong,YUAN Dan,HE Tai-ping. Antigenic and Genetic Characteristics of Hemagglutinin Gene of H1N1 Influenza Viruses Circulating in Human[J]. Practical Preventive Medicine, 2010, 17(12): 2386-2388 |
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| Authors: | WANG Hua-hong YUAN Dan HE Tai-ping |
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| Affiliation: | WANG Hua-hong,YUAN Dan,HE Tai-ping(Yueyang Municipal Center for Disease Control and Prevention,Yueyang 414000,Hunan,China) |
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| Abstract: | Objective To isolate the influenza A(H1N1) virus in vitro,and to explore the biological characteristics of the hemagglutinin gene. Methods Nasopharyngeal secretion was collected from the influenza virus sufferers and viral isolation was carried out by MDCK cells.The total RNA was extracted.One step RT-PCR was utilized to amplify the ORF fragment of HA gene with specific primers,and then the product was cloned to pUCm-T vector and pUCm-T/HA was transferred to Escherichia coli XL1-Blue.After the positive clon... |
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| Keywords: | Influenza A Viral isolation HA Biological characteristics |
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