鹅副黏病毒核酸探针检测方法的建立与应用

Establishment and application of digoxigenin-labeled nucleic acid probe for GPMV

根据GenBank中已发表的鹅副黏病毒F基因序列,设计合成1对引物,利用RT-PCR扩增出1条与目的片段大小一致,约856bp的基因片段,回收并纯化此PCR产物。用随机引物法合成cDNA,地高辛标记,建立了地高辛标记探针检测鹅副黏病毒的方法。该探针能与鹅副黏病毒核酸发生特异性杂交,最低检出限量为3ng/L;而与H9N2亚型禽流感病毒、鹅细小病毒、大肠杆菌等核酸杂交均为阴性。对疑似鹅副黏病毒感染病变组织检测结果表明,气管、肺脏、脾脏、肝脏均可检测出鹅副黏病毒,以气管的检出率为最高。该研究为鹅副黏病毒的诊断和流行病学调查提供了一种可靠的方法。

According to the sequence of the F gene published in GenBank,a pair of primers were designed for amplifying the 856 bp fragment in RT-PCR.The PCR product was labeled with digoxigenin as cDNA probe for detecting the GPMV.The result of the detection showed that the RNA of the GPMV were positive,the other virus and bacteria were negative.The sensitivity result showed that as few as 3 μg/L of GPMV was detected by DIG-labeled probe.The detection results for clinical samples tissue demonstrated that GPMV could be...