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。

关键词: 地高辛标记的核酸探针, 猪流感病毒, 检测

Abstract: The matrix gene's conserved region of swine influenza virus (SN) was labelled with digoxigenin as a cDNAprobe. The DIGlabelled fragment was tested by dot-blot hybridization with the cDNA of Hl subtype SN,H3 subtypeSN,PRRSV,CSFV and the DNA of PCV2,PRV for specificity. The result showed that the Hl and H3 subtype SN werepositive. As little as 5 pg cDNA of H3 subtype SN could be detected by the DIGlabelled probe. 35 suspicious sampleswere detected by the methods of nucleic acid probe and RT-PCR,and 9 positive cases were detected by each method. Theresult of dot-blot hybridization was in accordance with that of RT-PCR.It showed that the probe had the high specificityand sensitivity and could be used in the field of clinical diagnosis and epidemic investigation.

Key words: DIGlabelled nucleic acid probe, Swine influenza virus, Detection

中图分类号: 

• S852.65