H3亚型猪流感病毒HA基因核酸探针的制备与应用  

Preparation and Application of HA Gene Nucleic Acid Probe of H3 Subtybe Swine Influenza Virus

作　　者：吕翠[1] 马小明[1] 尹燕博[1] 温建新[1] 单虎[1] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109

出　　处：《西北农业学报》2009年第3期6-10,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金："十一五"国家科技支撑计划(2006BAD06A11);农业部"948"项目(2004-Z40)

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。The hemagglutinin gene's conserved region of H3 subtype swine influenza virus(SIV) was labelled with digoxigenin as a cDNA probe.The DIG-labelled fragment was tested by dot-blot hybridization with the cDNA of H3 subtype 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/μL cDNA 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.

关 键 词：地高辛标记 核酸探针 H3亚型 猪流感病毒 检测 

分 类 号：S854.43[农业科学—临床兽医学]