猪水肿病PCR快速检测试剂盒的研制

Development of Rapid PCR Detection Kit for Porcine Edema Disease

实验在大肠杆菌产生的志贺样毒素2型变异体(Shiga-like toxinⅡe,Stx2e)研究的基础上,设计了一对特异性引物,通过优化,建立了以PCR为基础,高效快速检测猪水肿病的PCR试剂盒。该试剂盒扩增的阳性条带为172bp的特异性DNA片段,最低检出菌体量为120CFU,在-20℃至少可保存12个月,重复性良好。应用猪水肿病PCR快速检测试剂盒对22份新鲜样本进行了检测,结果显示,其中的5个样本含Stx2e阳性大肠杆菌,PCR检测结果与剖检、细菌学和生化检验结果相一致。

Shiga-like toxin Ⅱ e（Stx2e） is responsible for edema disease of swine. A pair of specific primers was designed based on the research of Stx2e and the polymerase chain reaction （PCR） kit was constructed to detect edema disease in swine. The PCR kit could amplify one single 172 bp DNA band from Stx2e gene specifically. The sensitivity of the PCR kit was 120 CFU of bacteria and could be stored over 12 months at -20℃. 22 fresh pig samples was detected by the PCR kit and the result showed/ive pigs contain Stx2e gene, which were coincided with their anatomized and histological diagnosis.