摘要
目的建立鉴定幽门螺杆菌(H.pylori)cagA,vacA基因型别的多重聚合酶链反应(PCR)方法,并对90例H.pylori临床分离株的cagA,vacA型别进行鉴定。方法将鉴定cagA基因的引物P1,P2及鉴别vacA基因型别的引物VAG-F,VAG-R,VA1-F,VA1-R,SS1-F加入同一反应管中,一次性扩增出2～4条特异性DNA片段,并判定H.pylori cagA,vacA基因型别。结果建立了鉴定H.pylori cagA,vacA基因型别的多重PCR系统,其灵敏度为300CFU。90例H.pylori菌株中,82例(91.1％)含cagA基因;vacA基因信号区sla,slb和s2型分别为46例(51.1％),36例(40.0％)和8例(8.90％);中间区ml和m2型分别为46例(51.1％),44例(48.9％)。82例sl型H.pylori菌株中79例(96.3％)为cagA^+,8例s2型H.pylori菌株中2例(25.0％)为cagA^+,其差异有显著统计学意义(X^2=38.39,P<0.001)。结论建立的鉴定H.pylori cagA,vacA基因型别的多重PCR方法具有简便、快速、特异、经济的特点。西安地区人群感染的H.pylori菌株绝大多数为cagA^+菌株,H.pylori vacA基因的sl型(特别是sla型)和cagA基因的出现密切相关。
AIM To establish a multiplex polymerase chain reaction(PCR) method for determination of Helicobacter pylori (H. pylori ) cagA, vacA genotypes and to type cagA, vacA genotypes of 90 H. pylori isolates. METHODS In single PCR tube, 2-4 specific DNA bands were concurrently amplified with 7 primers (primers P1 and P2 for cagA gene, primers VAG-F, VAG-R, VA1-F, VA1-R and SS1-F for vacA genotypes). According to specific products, H. pylori cagA and vacA genotypes were dertermined. RESULTS A multiplex PCR method was established to dertermin H. pylori cagA, vacA genotypes. The sensitivity of this system was about 300CFU. Of 90 H. pylori isolates, 82 (91.1%) were cagA positive. H. pylori strains of s1a, s1b and s2 vacA genotypes were 46 (51.1% ), 36(40.0% ) and 8(8.9% ) respectively. H. pylori strains of m1 and m2 types were 46(51.1%) and 44(48.9%). In total, 96.3% of type vacA s1 H. pylori strains and 25.0% of type vacA s2 H. pylori strains were cagA positive ( X^2 = 38.39, P<0.001 ). CONCLUSION This multiplex PCR system is a highly specific, rapid, simple and economical method. H. pylori strains from Xi'an area were almost cagA positive. Thepresense of the cagA gene was closely associated with type vacA s1 (especially type s1a) gene of H. pylori strain.
出处
《世界华人消化杂志》
CAS
2001年第1期34-38,共5页
World Chinese Journal of Digestology
关键词
幽门螺杆菌
聚合酶链反应
胃粘膜
基因型
基因表达
Helicobacter pylori
polymerase chain reaction
gastric mucosa
genotype
gene expression
gene amplification
