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目的考察致泻性大肠埃希氏菌试剂盒对目标菌检测的准确性、特异性、抗干扰性能。方法①致泻性大肠埃希氏菌PCR检测:按照试剂盒说明书进行PCR反应体系配制和PCR程序扩增,对PCR产物采用Qiaxcel Advanced全自动核酸蛋白分析仪进行电泳分离,参照试剂说明书对目标菌评定;②通过对与目标菌生境相似、高频并存的非目标菌的混合模板进行检测,对试剂盒的特异性进行评估;③通过对目标菌和非目标菌的混合模板进行检测,对试剂盒的抗干扰能力进行评估。结果该试剂盒对5种致泻性大肠埃希氏菌的特征基因均能正确扩增;体系中加入目标菌的同时,加入大量的非目标菌,目标片段仍有很好的扩增,除检出目标菌特异条带,无非特异性的条带产生。结论对相应的目标菌都能给出准确的鉴定结果,特异性良好,且生境相似高频并存的非目标菌对目标菌的检测不产生干扰。
Objective To investigate the accuracy, specificity and anti-interference ability of the diarrhea-causing Escherichia coli kit for detecting the target bacteria. Methods ① diarrhea Escherichia coli PCR detection: according to kit instructions for PCR reaction system and PCR amplification program, PCR products using Qiaxcel Advanced automatic nucleic acid protein analyzer electrophoresis separation, reference to reagent instructions for the target bacteria Assessment of the specificity of the kit through the detection of the mixed template of non-target bacteria with similar habitat and high frequency coexistence with the target bacteria; (3) through the detection of the mixed template of target bacteria and non-target bacteria, Anti-interference ability of the box is evaluated. Results The kit was able to correctly amplify the characteristic genes of all 5 kinds of diarrhea-causing Escherichia coli. In the system, a large number of non-target bacteria were added into the system, and the target fragment was still amplified well. Detection of target bacteria specific bands, no non-specific bands. Conclusion The corresponding target bacteria can give accurate identification results, good specificity, and non-target bacteria coexisting with similar high-frequency habitats do not interfere with the detection of the target bacteria.