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Detection of Helicobacter pylori DNA by a simple stool PCR method in adult dyspeptic patients

机译:简单粪便PCR法检测成人消化不良患者的幽门螺杆菌DNA

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Introduction. Helicobacter pylori is the major agent causing peptic ulcer, gastric cancer and mucosa-associated lymphoid tissue (MALT) gastric lymphoma. A simple stool polymerase chain reaction (PCR) method was performed and compared with the gold standards for the diagnosis of H. pylori infection.Material and methods. A total of 54 adult patients (mean age, 46.41 +/- 13.12 years) with dyspeptic symptoms from Gastroenterology at Dokuz Eylul University Hospital between May and November 2003 were included. Two antrum and corpus biopsies were taken from each patient. Infection by H. pylori was defined as positivity and negativity of the gold standards. DNA extraction of stool specimens was done using QIAamp DNA Stool Mini Kit (QIAGEN) and PCR conditions included amplification and reamplification steps using the H. pylori ureA gene specific primers (HPU1, HPU2) and were visualized on 1% agarose gel stained with ethidium bromide.Results. Forty-six of 54 patients (85.2%) were diagnosed positive and eight (14.8%) were negative for H. pylori infection by the gold standard methods. Thirty-two patients were positive (59.3%) and 22 of them (40.7%) were detected negative by stool PCR method. The stool PCR method and gold standard methods showed a statistical difference for the detection of H. pylori infection (p < .0001). Sensitivity, specificity, likelihood ratio, and positive and negative predictive values were 65.22%, 75%, 2.61%, 93.75%, and 27.7%, respectively.Discussion. The PCR on the stool specimens resulted as being a very specific test. We suggest that a simple stool PCR method that we developed can be used to detect H. pylori, virulence genes, and in drug resistance studies either first line diagnostic methods in the laboratory or in the clinical management of dyspeptic patients.
机译:介绍。幽门螺杆菌是引起消化性溃疡,胃癌和与粘膜相关的淋巴样组织(MALT)胃淋巴瘤的主要药物。进行了简单的粪便聚合酶链反应(PCR)方法,并将其与诊断幽门螺杆菌感染的金标准进行了比较。材料和方法。在2003年5月至2003年11月之间,共有54名成年人(平均年龄为46.41 +/- 13.12岁)患有消化不良症状,其消化不良症状来自Dokuz Eylul大学医院的胃肠病学。每位患者均进行了两次胃窦和体活检。幽门螺杆菌的感染被定义为金标准的阳性和阴性。使用QIAamp DNA Stool Mini Kit(QIAGEN)进行粪便样本的DNA提取,PCR条件包括使用幽门螺杆菌ureA基因特异性引物(HPU1,HPU2)进行的扩增和再扩增步骤,并在1%溴化乙锭琼脂糖凝胶上观察结果。通过金标准方法,诊断为幽门螺杆菌感染的54例患者中有46例(85.2%)被诊断为阳性,而8例(14.8%)为阴性。粪便PCR法检测出32例阳性(59.3%),其中22例(40.7%)被检测为阴性。粪便PCR方法和金标准方法在检测幽门螺杆菌感染方面显示出统计学差异(p <.0001)。敏感性,特异性,似然比以及阳性和阴性预测值分别为65.22%,75%,2.61%,93.75%和27.7%。粪便标本的PCR结果是非常特殊的测试。我们建议,我们开发的一种简单的粪便PCR方法可用于检测幽门螺杆菌,毒力基因,以及在耐药性研究中,无论是实验室中还是消化不良患者的临床管理中的一线诊断方法。

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