球形幽门螺杆菌致细胞空泡变性毒力研究

摘要

Aim To reveal the virulence variation related to vacuolating cytotoxicity of coccoid Helicobacter pylori . Methods The coccoid Heicobacter pylori, which was transfonmed from the bacterium with cagA gene and vacA gene by mesans of prolonged incubation and antibiotics with subinhibitory concentration, was detected the vacuolating cytotoxin to Hela cells, analysed the differences of protein between the verity strain snd its prototype by using SDS-PAGE, studied mutation of the vacA gene and cagA gene by means of PCR and PCR-SSCP.Results and Conclusions. The vacuolating cytotoxicity, the proteins with molecular weights above 74KD, especially 125KD, were decreased, no deletion was found in 259bp smplification fragments from vacA gene and 349bp amplificstion fragments from cagA gene but point mutation was existed in vacA gene of coccoid Hp.%目的　揭示球形幽门螺杆菌致细胞空泡变性毒力的变异情况。方法　采用延期培养和加亚抑菌浓度抗生素，对细胞毒相关蛋白基因阳性(cagA+)，和细胞空泡毒素基因阳性(vacA+)的高毒株幽门螺杆菌(Hp)进行球形诱变，进而检测该Hp至Hela细胞空泡变性的毒力，并用SDS-PAGE、PCR及PCR-SSCP技术，分析球形Hp蛋白产物及决定细胞空泡变性的cagA和vacA基因的变异情况。结果　与结论　球形Hp致细胞空泡毒力减弱，74KD以上的蛋白含量减少，特别是125KD条带尤为明显。259bp的vacA基因片段和349bpcagA基因片段未发生缺失，但两种方法　诱变的球形HpvacA基因均存在点突变。