【摘要】 用已建立的RT-PCR方法对采集的疑似猪流行性腹泻病猪的粪便样品进行检测，选用Vero细胞分离培养检测猪流行性腹泻病毒(PEDV)阳性的样品，连续传3代后用RT-PCR方法鉴定，对其S1基因进行序列测定和遗传进化分析，并用Vero细胞对分离毒体外培养所需的接毒时间、收毒时间、接毒剂量及胰蛋白酶浓度进行试验，筛选出该株PEDV体外培养最佳条件。结果表明，该分离株为G2b型，在Vero细胞增殖最佳接毒时间为培养24 h细胞长成单层时，最佳收毒时间为接毒后48 h,最佳接毒剂量为10%,最佳胰蛋白酶浓度为15μg/mL,研究结果可为PEDV的分离培养及提高病毒效价提供参考。更多还原

【Abstract】 In order to understand the molecular variation and in vitro culture characteristics of PEDV in Shandong province, in this experiment, the established RT-PCR method was used to detect the fecal samples of pigs suspected of epidemic diarrhea, and Vero cells were selected for virus isolation.After three generations of continuous transmission, it was identified by RT-PCR,and its S1 gene was sequenced and its genetic evolution was analyzed.The inoculation time, harvest time, inoculation dose and trypsin concentration of the PEDV culture were tested with Vero cells to screen out the optimal conditions for the proliferation of PEDV.The results showed that the phylogenetic tree analysis of S1 gene showed that the isolate was G2b type.The optimal inoculation time of PEDV in Vero cell proliferation was 24 hours after cell culture, and the optimal harvest time was 48 hours after infection.The optimal inoculation dose was 10%,and the optimal trypsin concentration was 15 g/ml.The results can provide reference for the isolation and culture of PEDV and the improvement of virus titer.更多还原