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目的建立环境水体中霍乱弧菌的二次PCR检测方法。方法对霍乱弧菌的外环境水监测样本进行PCR检测;确定弱电泳条带或无电泳条带的稀释度,将第一次PCR的产物梯度稀释后进行二次PCR。采用琼脂凝胶糖电泳进行分析。结果对378件环境水样分别进行一次、二次PCR检测,有4件水样经两种方法检测均为阳性,有6件水样二次PCR检测为阳性。一次PCR检测霍乱弧菌的阳性率[1.06%(4/378)]低于二次PCR[2.65%(10/378)],差异有统计学意义(P<0.05)。结论二次PCR操作简便快速、检出率高、费用低廉,适用于环境水样中霍乱弧菌的快速监测。
Objective To establish a second PCR detection method for Vibrio cholerae in environmental water. Methods Vibrio cholerae external environmental water monitoring samples were detected by PCR. The dilution of weak electrophoresis bands or no electrophoresis bands was determined. The products of the first PCR were diluted and then subjected to secondary PCR. Agarose gel electrophoresis analysis. Results A total of 378 environmental water samples were subjected to one and two PCR tests respectively. There were 4 water samples tested positive by both methods, and 6 water samples were positive by secondary PCR test. The positive rate of Vibrio cholerae by one PCR was lower than 1.06% (4/378) and lower than that by PCR (2.65% (10/378)). The difference was statistically significant (P <0.05). Conclusion The secondary PCR is simple and rapid, with high detection rate and low cost. It is suitable for the rapid monitoring of Vibrio cholerae in environmental water samples.