聚合酶链反应检测空肠和结肠弯曲菌的核酸

Specific detection of campylobacter jejuli and campylobacter coli by using polymerase chain reaction

为了解急性腹泻病人空肠和结肠弯曲菌的感染率，建立了空肠和结肠弯曲菌的ＰＣＲ检测方法。引物设计在结肠弯曲菌鞭毛Ａ基因高度保守区，扩增产物为４５０ｂｐ。结果显示该方法仅在空肠和结肠弯曲菌ＤＮＡ有扩增产物出现，最小检测量达５０ｆｇＤＮＡ，而其它细菌如幽门弯曲菌，亲水气单胞菌，豚鼠气单胞菌，副溶血弧菌和产肠毒素性大肠埃希菌均未见扩增产物。检测急性腹泻患者粪便１００份，１１例为阳性。

A PCR method was established to detect the infection rate of C. jejuli and C. coli in the patient suffering from acute diarrhea. The designed primer located in the conservative region of flagellin a gene of C. coli. The amplifited product was 450bp. The experiments of sensitivity,specificity and identification of the amplified product have been carried out. The results showed that,this assay could detect as little as 50fg of C.jejuli DNA;only amplified product of C.jejuli DNA could be viewed under UV light,and none of amplified products of helicobacter pylori,aeromonas hydrophila,aeromonas caviae,vibrio parahemolyticus and enterotoxigenic E. coli could be found. The amplificed product have been identified with target DNA and Marker.A handred stool samples were examined and eleven stool samples were positive. Meanwhile routine culture of stool were all negative.