空肠弯曲菌多重PCR快速检测方法建立的研究

STUDY ON MULTIPLEX PCR ASSAY FOR RAPID DETECTION OF CAMPYLOBACTER JEJUNI

[目的]建立一种能够快速检测鉴别样品中的空肠弯曲菌的多重PCR方法。[方法]根据空肠弯曲菌hipO、mapA、23S等基因序列,利用Primer Premier 5.0和Oligo 6软件设计了多对特异性引物,经过优化筛选出最佳引物组合,再对引物的浓度、Mg2+浓度、退火温度等条件进行优化后,建立了从样品中快速检测鉴别空肠弯曲菌的多重PCR方法。[结果]实验结果显示本方法的特异性较强,灵敏度高,达到15 cfu/ml。[结论]多重PCR方法对于样品中空肠弯曲菌的检测实验周期短,灵敏度高,操作简便,适宜推广应用于各卫生单位日常对样品中空肠弯曲菌污染的快速检测。

[Objective] To set up a multiplex PCR assay for rapid detection of Campylobacter jejuni from food samples. [Methods] Based on the sequence of hipO gene, mapA gene and 23S gene, some couples of specific primers were designed, with the utilization of primer select software of Primer Premier 5.0 and Oligo 6. Then the most specific primers pairs were chosen to carry out optimization of the concentration of primer, Mg^2＋ and Tm℃, etc. The assay of multiplex PCR assay for detection of Campylobacter jejuni from food samples was established. [ Results] The results showed that this assay possessed high specificity and sensibility for detection of Campylobacter jejuni, which achieved 15cfu/ml. [Conclusion] This assay of multiplex PCR can detect Campylobacter jejuni from food samples. It is low time consuming, convenient and high efficient. And it should be utilized in our daily detection work.