空肠弯曲菌实时荧光定量PCR检测方法的研究

目的建立一种快速廉价的检测空肠弯曲菌的实时荧光定量PCR方法。方法根据空肠弯曲菌flaA、gyrA基因设计引物,建立空肠弯曲菌实时荧光定量PCR检测方法。结果以flaA基因引物对空肠弯曲菌DNA进行实时荧光定量PCR,扩增曲线呈S形指数增长,大肠埃希菌等对照菌扩增阴性。方法的灵敏度为10CFU/ml菌悬液。结论建立的空肠弯曲菌荧光定量PCR检测方法具有快速、简便、准确等特点,具有一定的使用价值。

Establishment of real-time fluorescent PCR for detection of Campylobacter jejuni

Objective To establish a rapid and accurate method for detection of Campylobacterjejuni by real-time fluorescent PCR assay. Methods According to the open reading frame （ORF） of sequence of flaA gene, gyrA gene, several couples of specific primers were designed. Reaction system and determination approach of real-time fluorescent PCR were established for detection of C. jejuni. Results The flaA primer pairs were used in real-time PCR assay for detection of C. jejuni, the fluorescent amplification curves were observed as a typical ＂S＂ curve, other bacteria were also tested and no amplification was observed. It was found that the sensitivity of this assay was 10 CFU/ml. Conclusion This real-time PCR assay is fast, simple, sensitive and specific for the purposes of detection of C. jejuni.