慢性乙型肝炎患者肝组织中螺杆菌菌属特异性16SrRNA基因及HP特异性基因分析

目的对慢性乙型肝炎患者肝组织进行螺杆菌菌属特异性16SrRNA基因及幽门螺杆菌（HP）特异性基因检测,探讨螺杆菌在慢性乙型肝炎发生、发展中的作用。方法对56例行肝穿刺活检的慢性乙型肝炎患者的肝组织应用针对螺杆菌菌属特异性16SrRNA基因的通用引物进行基因扩增及微需氧分离培养,并对螺杆菌菌属特异性16SrRNA基因阳性者进一步应用HPcagA、vacA和glmM基因特异引物进行扩增。结果对56例慢性乙型肝炎患者的肝组织DNA应用螺杆菌菌属特异性16SrRNA基因通用引物进行扩增,共有35例患者的肝组织中发现了螺杆菌菌属特异性16SrRNA基因,其中肝硬化组（72.7%）和原发性肝癌组（87.5%）的检出率明显高于慢性肝炎组（42.3%,P〈0.05）,而慢性肝炎组中随着炎症评分的增加,检出率亦增加。对这35例螺杆菌菌属特异性16SrRNA基因阳性的肝组织DNA进一步应用HPcagA、vacA和glmM基因特异性引物进行扩增,结果证实有21例为HPDNA。所有肝组织经微需氧分离培养均未发现可疑菌落生长。结论慢性乙型肝炎患者肝组织中除存在HPDNA外,可能还存在其他螺杆菌DNA。螺杆菌在慢性乙型肝炎向肝硬化和肝癌的发展中可能发挥致病作用。

Identification of Helicobacter genus-specific 16SrRNA and H. pylori specific genes in liver samples in patients with the chonichepatitis B

Objective To detect Helicobacter genus-specific 16SrRNA and H.pylori specific genes in liver samples in patients with chonic hepatitis B,and to investigate whether Helicobacter species play a role in the development of the chronic hepatitis B.Methods Helicobacter were sought by PCR using primers for the 16SrRNA and histology culture in the liver samples from 56 patients with chronic hepatitis B,and H.pylori cagA、vacA and glmM genes were further amplified by specific primers amplification in the samples with Helicobacter genus-16SrRNA positive. Results Helicobacter genus-specific 16SrRNA was found in 35 of 56 samples,and the detection rate in the patients with HCC,chronic cirrhosis were significantly higher than that in the patients with chronic hepatitis （87.5%,72.7% versus 42.3%,P﹤0.05）. In the patients with chronic hepatitis,the positive rate increased with the inflammation score. Of the 35 patients,21 were detected positively for H.pylori cagA、vacA and glmM genes by PCR. The other 14 samples were sequenced,and found to have 99.2% similarity to the 16SrRNA of H.hepaticus and 98.7% similarity to the 16SrRNA of H.rodentium,96.9% similarity to the 16SrRNA of H. pullorum,95.2% similarity to the 16SrRNA of H.bilis.Conclusion H.pylori DNA and other helicobacter DNA coexisted in the liver samples of patients with chronic hepatitis B,and they might play a synergistic role in the development of chronic hepatitis to cirrhosis and HCC.