犬钩端螺旋体PCR检测方法的应用

Application of PCR Method for Detection of Canine Leptospira

采用荷兰皇家热带病研究院WHO钩体病参考中心设计的一对引物进行普通PCR检测。经反应条件与体系优化,对犬钩端螺旋体与霍乱弧菌,伤寒杆菌,沙门氏菌,产毒性大肠杆菌,致病性大肠杆菌,痢疾,溶藻菌,非O1、O139霍乱弧菌,副溶血性弧菌,O157菌进行检测,除犬钩端螺旋体以外均为阴性,表明钩端螺旋体PCR检测方法与以上菌株无交叉反应,证明了该钩端螺旋体PCR检测方法具有特异性;双链DNA灵敏度检测结果显示PCR方法可以检测到的核酸浓度最低为3×10^(3)拷贝。此结果证明该犬钩端螺旋体PCR检测方法具有较好的特异性和灵敏度,可用于犬钩端螺旋体样品的检测。

A pair of primers designed by WHO reference center of leptospirosis of the Royal Institute of Tropical Diseases in the Netherlands was used to detect leptospirosis.After optimization of reaction conditions and system,the detection of Canine Leptospira with Vibrio cholerae,Salmonella typhi,enterotoxigenic Escherichia coli,pathogenic Escherichia coli,dysentery,alginolytic bacteria,non-O1,O139 Vibrio cholerae,Vibrio paralyticus and O157 were conducted.The results showed that there was no cross reaction between the PCR detection method and the above strains.The results of double strand DNA sensitivity test showed that the PCR method could detect 3×10^(3) copies of nucleic acid with good specificity and sensitivity,and thus could be used for the detection of Canine Leptospira samples.