猪流感病毒M基因核酸探针的制备与应用  

Preparation and Application of M Gene Nucleic Acid Probe of Swine Influenza Virus

作　　者：吕翠[1] 马小明[1] 尹燕博[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：《华北农学报》2009年第1期87-92,共6页Acta Agriculturae Boreali-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)标记猪流感病毒(SIV)M基因的保守片段制成核酸探针,与H1及H3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(PRV)的DNA进行斑点杂交,以检测探针的特异性,结果该探针仅与H1及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;敏感性试验显示,探针最低能检出约5 pg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9份阳性,与RT-PCR检测结果一致。本研究结果表明,建立的DIG标记的核酸探针特异性强,敏感性高,适合于对SI的诊断和流行病学调查。The matrix gene＇s conserved region of 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 H1 subtype SIV, H3 subtype SIV,PRRSV, CSFV and the DNA of PCV2,PRV for specificity. The result showed that the H1 and H3 subtype SIV were positive. As little as 5 pg cDNA of H3 subtype SIV could be detected by the DIG-labelled probe. 35 suspicious samples were detected by the methods of nucleic acid probe and RT-PCR, and 9 positive cases were detected by each method. 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：地高辛标记的核酸探针 猪流感病毒 检测 

分 类 号：S852.65[农业科学—基础兽医学]