Sen Nazime, Yilmaz Ozlem, Simşek Ilkay, Küpelioğlu Ahmet Ali, Ellidokuz Hülya
Department of Microbiology and Clinical Microbiology, Faculty of Medicine, Dokuz Eylül University, Inciralti 35340 Izmir, Turkey.
Helicobacter. 2005 Aug;10(4):353-9. doi: 10.1111/j.1523-5378.2005.00326.x.
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.
A total of 54 adult patients (mean age, 46.41 +/- 13.12 years) with dyspeptic symptoms from Gastroenterology at Dokuz Eylül 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.
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.
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.
幽门螺杆菌是导致消化性溃疡、胃癌和黏膜相关淋巴组织(MALT)胃淋巴瘤的主要病原体。我们采用了一种简单的粪便聚合酶链反应(PCR)方法,并与诊断幽门螺杆菌感染的金标准进行比较。
纳入2003年5月至11月在多库兹艾吕尔大学医院胃肠病科有消化不良症状的54例成年患者(平均年龄46.41±13.12岁)。从每位患者采集两块胃窦和胃体活检组织。幽门螺杆菌感染根据金标准的阳性和阴性来定义。粪便标本的DNA提取使用QIAamp DNA Stool Mini Kit(QIAGEN),PCR条件包括使用幽门螺杆菌ureA基因特异性引物(HPU1、HPU2)进行扩增和再扩增步骤,并在经溴化乙锭染色的1%琼脂糖凝胶上进行可视化分析。
通过金标准方法,54例患者中有46例(85.2%)被诊断为幽门螺杆菌感染阳性,8例(14.8%)为阴性。粪便PCR方法检测出32例阳性(59.3%),其中22例(40.7%)检测为阴性。粪便PCR方法与金标准方法在幽门螺杆菌感染检测方面存在统计学差异(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结果显示是一种非常特异的检测方法。我们建议,我们开发的这种简单的粪便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,无论是作为实验室的一线诊断方法还是用于消化不良患者的临床管理。