原位杂交检测鸡肿瘤病

Detection of Chicken Neoplastic Diseases by in Situ Hybridization

本研究旨在运用原位杂交检测方法检测鸡的肿瘤病。根据GenBank中登录的MDV、ALV-J和REV参考毒株序列,设计并合成3对引物,经PCR方法扩增出相应的片段,并进行了克隆测序。将测序正确的片段亚克隆至pSPT-18,经转录得到DIG标记的特异cRNA探针。将探针分别与3种肿瘤病阳性组织石蜡切片进行杂交,可看到阳性信号,特异性、敏感性较好。运用该方法对32份疑似肿瘤病料进行了检测,MDV、ALV-J和REV的阳性率分别为25.00%、18.75%和3.13%。结果提示,鸡肿瘤病在鸡场中仍广泛存在。

This experiment was conducted to using a method of in situ hybridization to detect chicken neoplastic diseases.Three pairs of primers were designed and synthesized based on the published sequences of MDV,ALV-J and REV.The PCR amplified products were subcloned into pSPT-18 plasmid to generate DIG-labeled cRNA probes targeting MDV,ALV-J and REV respectively.Positive sections was hybridized with the three probes and the results showed good specificity and sensitivity.Thirty-two suspected neoplastic tissue samples from different farms were processed as paraffin-embedded sections and hybridized with RNA probes above.Of these samples,MDV,ALV-J and REV positive rates were 25.00%,18.75% and 3.13% respectively.The study showed that chicken neoplastic diseases were still widely existed in the chicken farms.