PCR、斑点杂交及间接免疫荧光试验对鸡传染性贫血病临床诊断的应用比较

Application and Comparison of the PCR, Dot blot hybridization Assay and IIFA for Diagnosis of Chicken Anemia Virus

应用能特异性扩增出鸡传染性贫血病毒（ Ｃ Ａ Ｖ） ０５８ ｋｂ Ｄ Ｎ Ａ 的已知引物，对江苏某地区疑为 Ｃ Ａ Ｖ感染的 １５～３０ 日龄病鸡的肝 Ｄ Ｎ Ａ 样品进行了 Ｐ Ｃ Ｒ 扩增。结果，在被检的 ２０ 份样品中，有 ６ 份为 Ｐ Ｃ Ｒ 阳性，阳性率 ３０％ 。利用地高辛标记的 ０８５ ｋ ｂ 的 Ｃ Ａ Ｖ 核酸探针对相同样品进行斑点杂交，结果与 Ｐ Ｃ Ｒ 扩增相同。对应的病鸡血清经间接免疫荧光试验（ Ｉ Ｉ Ｆ Ａ）发现，有 ７ 份为 Ｃ Ａ Ｖ 抗体阳性，与 Ｐ Ｃ Ｒ 扩增结果的符合率为 ７７％ 。初步结果表明，江苏某地区存在 Ｃ Ａ Ｖ 感染， Ｉ Ｉ Ｆ Ａ 与 Ｐ Ｃ Ｒ

The specific fragments (0.58 kb) of the chicken aneamia virus (CAV) DNA were amplified by the polymerase chain reaction (PCR). The CAV DNA was detected by PCR and dot blot hybridization assay using the 0.85 kb PCR products labeled with Digoxigenin as probes. Using the two methods, the tissue DNA samples of 15 30 day old different sick chickens collected from the areas of Jiangsu province were detected and 6 out of 20 chicken flocks were positive (the positive rate was 30%). For the serums of the same samples, 7 out of 20 (35%) were positive by the indirect immunofluorescence assay (IIFA). The result of the IIFA compared with the PCR/dot blotting showed 83% coincidence rate. The preliminary investigation showed that the CAV infection is prevalent in the areas of Jiangsu province and that there are some different results between the IIFA and PCR/dot blotting.