致泻性大肠埃希氏菌多重PCR检测试剂盒的质量评估

Evaluation Study of multiplex PCR assay for detection Diarrheagenic Escherichia coli

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 Diarrheagenic Escherichia coli multiplex PCR assay for detection of target bacteria, specificity, anti-jamming performance evaluation Method Diarrheagenic Escherichia coli by multiplex PCR respectively ac- cording to a variety of target bacteria kit instructions PCR mixture and PCR amplification procedures, the PCR products were Qiaxcel Advanced automated nucleic acid protein electrophoretic separation analyzer,reference reagent instruc- tions on the target bacteria identification; With the target bacteria by habitat similar to the coexistence of non- target or- ganisms and high frequency hybrid template to detect Diarrheagenie Escherichia coli on Diarrheagenic Escherichia coli multiplex PCR assay to assess the specificity;Through the target and non-target bacteria bacteria mixed template to de- tect foodborne pathogens multiplex PCR assay to assess anti-jamming capability Result Target bacteria added to the system at the same time, adding a large number of non- target organisms, target amplification fragments are still very good, except for the detection of target bacteria specific bands, no nonspecific bands produced .Conclusion The cor- responding target bacteria can give an accurate identification results, specificity is good, and the coexistence of habitat similar to high-frequency non-target bacteria on the detection of target bacteria do not produce interference.