荧光PCR法检测人胃组织中幽门螺旋杆菌核酸的实验研究

Real-time quantitative PCR for detection of Helicobacter pylori in human gastric tissue

目的探讨荧光定量PCR方法检测人胃组织中幽门螺杆菌的意义。方法收集399例上消化道疾病患者的胃粘膜活检标本。其中食管炎8例,食管溃疡5例,食管癌15例,胃炎301例,胃溃疡30例,胃癌5例,十二指肠炎4例,十二指肠溃疡29例,十二指肠癌2例。核酸测序作为金标准方法,分析荧光定量PCR方法检测胃粘膜幽门螺杆菌的价值。结果荧光PCR法检测胃组织中,阳性数206份,阳性率51.6%,其中胃炎阳性151例(50.2%,151/301),其余各组阳性率与金标准一致,阴性数193份,阴性率48.4%。Mcnemar检验示,配对比较显示金标准法与荧光PCR法无统计学差异(P=1.000)。以测序法为金标准,荧光PCR法的灵敏度为99.5%,特异性为98.5%,准确度为99.25%,Kappa值=0.985(P=0.009)。结论荧光PCR法能够快速、准确检测人胃组织幽门螺杆菌。

Aim To explore the real-time quantitative PCR-based assay to detect Helicobacter pylori in human gastric tissue.Methods 399 human gastric biopsy samples from patients with upper digestive disease were collected,including esophagitis（n=8）,oesophageal ulcer（n=5）,esophageal carcinoma（n=15）,gastritis（n=301）,gastric ulcer（n=30）,gastric carcinoma（n=5）,duodenitis（n=4）,duodenal ulcer（n=29）and duodenum carcinoma（n=2）.Based on the ＂golden＂ method,PCR-based assay value for detection of Helicobacter pylori in human gastric tissue was evaluated by Mcnemar test and Kappa test.Results By the PCR assay,206 gastric samples（51.6%）were positive to Helicobacter pylori.Helicobacter pylori positivity was 50.2% in patients with gastritis.The sensitivity of PCR assay was 99.5% and the specificity was 98.5%.The agreement test showed the kappa value was 0.985（P=1.000）.Conclusion Real-time-PCR method is a rapid and sensitive technique to detect Helicobacter pylori human gastric tissue.