| H3亚型猪流感病毒HA基因核酸探针的制备与应用 |
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| 引用本文： | 吕翠,马小明,尹燕博,温建新,单虎.H3亚型猪流感病毒HA基因核酸探针的制备与应用[J].西北农业学报,2009,18(3):6-10. |
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| 作者姓名： | 吕翠 马小明 尹燕博 温建新 单虎 |
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| 作者单位： | 青岛农业大学,动物科技学院,山东青岛,266109 |
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| 基金项目： | 国家科技支撑计划，农业部引进国际先进农业科学技术计划(948计划) |
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| 摘 要： | 以地高辛(DIG)标记H3亚型猪流感病毒(SIV)HA基因的保守片段制成核酸探针,与H3及H1亚型的猪流感病毒(SIV)的cDNA、猪呼吸与繁殖综合征病毒(PRRSV)的cDNA、猪圆环病毒Ⅱ型(PCV2)的DNA、猪瘟病毒(CSFV)cDNA、猪伪狂犬病病毒(PRV)的DNA进行斑点杂交,检测探针的特异性.结果显示,该探针仅与H3亚型的猪流感病毒(SIV)的cDNA杂交呈阳性,与其余病毒杂交呈阴性;敏感性试验结果显示,探针最低能检出约6 pg/μL的H3亚型的SIV RT-PCR产物.应用该探针对35份疑似SI临床病料进行检测,结果检出4份阳性病料,与RT-PCR检测结果一致.研究表明,建立的DIG标记的核酸探针特异性强,敏感性高,适合于对H3亚型SI的诊断和流行病学调查.
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| 关 键 词： | 地高辛标记 核酸探针 H3亚型 猪流感病毒 检测 |
| 收稿时间： | 2008/11/3 0:00:00 |
| 修稿时间： | 2009/1/5 0:00:00 |
Preparation and Application of HA Gene Nucleic Acid Probe of H3 Subtybe Swine Influenza Virus |
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| L&#; Cui,MA Xiaoming,YIN Yanbo,WEN Jianxin and SHAN Hu.Preparation and Application of HA Gene Nucleic Acid Probe of H3 Subtybe Swine Influenza Virus[J].Acta Agriculturae Boreali-occidentalis Sinica,2009,18(3):6-10. |
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| Authors: | L&#; Cui MA Xiaoming YIN Yanbo WEN Jianxin and SHAN Hu |
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| Institution: | Faculty of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China;Faculty of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China;Faculty of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China;Faculty of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China;Faculty of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China |
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| Abstract: | The hemag glutinin genes conserved region of H3 subtype swine influenza virus(SIV) was labelled with digoxigenin as a cDN A pro be.The DIG-labelled fragment was tested by dot-blot hybridization with the cDNA of H3 subty pe SIV,H1subtype SIV,PRRSV and the DNA of PCV2,PRV for specificity.The result showed that only the H3 subtype SIV was positive.As little as 6 pg/LcDNA of SIV could be detected by the DIG-labelled probe.35 samples were detected by the methods of nucleic acid probe and RT-PCR,and 4 positive cases were detected.The result of dot-blot hybridization was in accordance with that of RT-PCR.It showed that the probe had the high specificity and sensitivity and could be used in the field of clinical diagnosis and epidemic investigation. |
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| Keywords: | DIG-labelled Nnucleic acid probe H3 subtype Swine influenza virus Detection |
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