摘要
目的建立检测幽门螺杆菌（Ｈｐ）的更为敏感的聚合酶链反应（ＰＣＲ），并探讨它在Ｈｐ感染的诊断及根除效果判定中的应用．方法以一对合成的与Ｈｐ１６ＳｒＲＮＡ基因互补的寡核苷酸为引物（ＣＰ１／ＣＰ２），建立了从胃粘膜中检测Ｈｐ的ＰＣＲ反应，并与常规检测方法进行比较．结果ＰＣＲ方法检测Ｈｐ标准菌株及５０株临床分离菌株均产生５００ｂｐ片段，检出的最小ＤＮＡ量为０１ｐｇ，相当于１００个细菌细胞；所有１３株其它细菌及无Ｈｐ感染的人胃粘膜则无扩增产物出现．用该方法检测９６名初诊患者Ｈｐ感染情况及２１名药物治疗后患者Ｈｐ根除情况，并与胃粘膜活组织尿素酶试验、细菌培养及银染色方法比较，证实ＰＣＲ方法可以检出常规方法不能检出的少量Ｈｐ．结论ＰＣＲ是检测Ｈｐ的最为敏感的方法。
AIM To establish the polymerase chain reaction(PCR) method for the detection of H.pylori from gastric mucosa specimens and to study its value in diagnosing H.pylori infection and confirming H.pylori eradication. METHODS A synthetic oligonucleotide primer pair derived from the 16S rRNA gene was used to develop PCR for the detection of H.pylori from gastric mucosa specimens and the results were compared to that of routine methods. RESULTS A 500bp fragment was amplified in 2 H.pylori reference strains and 50 clinically isolated strains, whereas 13 other bacterial species and normal human gastric mucosa were found not to yield amplified products. PCR can detect as little as 0 1pg of DNA, corresponding to about 100 bacteria cells. By comparing PCR with urease test, culture and Warthin_Starry stain in 96 patients with upper gastrointestinal symptoms and 21 patients with H.pylori associated duodenal ulcer after treatment, it is demonstrated that PCR was more sensitive than routine methods for detecting small numbers of bacteria which could not be detected by the routine diagnostic tests. CONCLUSION PCR is a rapid, sensitive and specific method for the detection of H.pylori . It can be used to diagnose H.pylori infection and to confirm eradication of H.pylori after treatment.
关键词
胃粘膜
微生物学
幽门螺杆菌
聚合酶链反应
Gastric mucosa/microbiology Helicobacter pylori /genetics DNA, bacterial/analysis Polymerase chain reaction
