Diagnostic Performance of glmM Gene and Histological Stains for Detection of Helicobacter pylori in Gastric Biopsy from Patients Admitted to Wad Madani Teaching Hospital, Sudan

Helicobacter pylori is the microbial agent most responsible for gastro-duodenal ulcer and chronic gastritis, which can develop into carcinoma of the stomach. This study was performed in Wad Medani Teaching Hospital, Sudan to detect Helicobacter pylori in stomach samples, and evaluate the performance of the tests used, which were histological stains and PCR. Gastric biopsies were obtained from 105 referred patients during endoscopy, and fixed specimens examined by haematoxylin-eosin and Warthin-Starry silver stains, while DNA was extracted for glmM gene amplification. Epigastric pain was the most common symptom at 78% (82/105) and chronic gastritis recorded with 71% (68/105) of endoscopy results. Warthin-Starry silver stain gave 31% (33/105) as positive for Helicobacter pylori followed by glmM gene 27% (28/105) and haematoxylin-eosin 24% (25/105). The study indicated good performance of histological staining and high specificity of glmM gene in detection of Helicobacter pylori from gastric biopsies.

奶牛乳样和粪样中幽门螺杆菌PCR检测方法的建立及初步应用

【目的】建立一种特异、灵敏的牛源幽门螺杆菌(Helicobacter pylori,Hp)检测方法,为家畜感染幽门螺杆菌的流行病学调查鉴定提供支持。【方法】分别选取Hp的16SrRNA、UreA、glmM为靶基因设计特异性引物,以H.pylori动物模型适应菌株SS1株为标准菌株建立PCR检测方法,并进行PCR条件优化,运用所建立的最优PCR方法检测临床奶牛乳样和粪样中Hp的分布情况。【结果】PCR特异性试验结果显示,仅Hp SS1株能扩增出特异性条带,而对照菌株金黄色葡萄球菌、沙门氏菌、单核细胞增生李斯特氏菌、大肠杆菌及蜡样芽孢杆菌均无扩增条带。PCR灵敏度试验结果显示,16SrRNA、UreA、glmM基因在乳样中的最低检出浓度分别为101,103和103CFU/mL,在粪样中的最低检出浓度分别为101,104和104 CFU/mL。应用该PCR方法对采集自规模化养殖场及散养的共计41头奶牛的乳样和粪样进行检测,成功检出Hp DNA,靶基因16SrRNA、UreA和glmM序列与NCBI上所发表的J166和ATCC43504菌株序列的同源性均达98%以上。【结论】成功建立了特异、灵敏的牛源Hp的PCR检测方法,可以应用于临床奶牛乳样和粪样中Hp的检测。