应用聚合酶链式反应检测犬细小病毒

将具有犬细小病毒病典型症状的病犬肠溶物经ＳＤＳ－酚裂解法提取总ＤＮＡ，并以此ＤＮＡ为模板，通过人工合成的两条２０ｍｅｒ引物进行ＰＣＲ扩增，该对引物扩增的片段为犬细小病毒结构蛋白ＶＰ２基因中间１／３区段，ＰＣＲ产物经琼脂糖凝胶电泳，试验结果表明：用ＰＣＲ扩增检测犬细小病毒具有良好的特异性，应用该方法检测的２７份病犬肠溶物样品块获得了预计长度的ＤＮＡ片段（５３１ｂｐ），而健康犬的肠溶物样品ＰＣＲ结果为

Detection of Canine Parvovirus by Polymerase Chain Reaction

The total DNA was isolated from the enteric lysate of virus infected dog showing enteritis symptoms.A pair of primers,20 mer oligonucleotides,spanning to the mid 1/3 region of VP 2 gene of canine parvovirus were synthesized.The total DNA was used as the template for PCR amplification,and the products of PCR were identified by 1% agarose gel electrophoresis.The results show that the PCR has a good specificity,being able to amplify DNA of expected length (531 bp) from 27 infected enteric lysate samples.But all the uninfected enteric lysate samples give negative results.Sensitivity test of PCR indicated that the maximum dilution of total DNA which gives positive reaction is 10 -6 ,corresponding to 1.37 pg total DNA.This diagnostic technique is rapid and simple because the samples are directly assayed from virus infected enteric lysate and the results could be obtained within 24 hours.This method can be used for clinical detection of the canine parvovirus with high sensitivity and specificity.