Cağdaş Umut, Otağ Feza, Tezcan Seda, Sezgin Orhan, Aslan Gönül, Emekdaş Gürol
Mersin University Faculty of Medicine, Department of Medical Microbiology, Mersin, Turkey.
Mikrobiyol Bul. 2012 Jul;46(3):398-409.
Helicobacter pylori is reported as the etiological agent of gastritis, gastric and duodenal ulcer, gastric adenoid carcinoma and mucosa-associated lymphoid tissue lymphoma. In the diagnosis of H.pylori infections invasive (culture, histopathological examination, rapid urease test and molecular tests) and non-invasive (urea breath test, serological tests, stool culture and stool antigen/nucleic acid tests) methods may be used. Clarithromycin, amoxicillin and combination of metronidazole and protonpump inhibitor or ranitidine bismuth citrate triple treatment protocol is applied in order to treat and eradicate the infection. However, increasing rates of antibiotic resistance among H.pylori strains reduces the success of eradication therapy. The aim of this study was to investigate the presence of H.pylori in the gastric antral biopsy specimens and to determine the antimicrobial resistance of the isolates. A total of 149 gastric antral biopsy specimens obtained from patients (age range: 17-83 years; 73 were male) who admitted to Mersin University Faculty of Medicine Department of Internal Medicine Gastroenterology clinic with dyspeptic complaints were included in the study. H.pylori presence was investigated by culture, polymerase chain reaction (PCR) and urease test from gastric biopsy specimens, and H.pylori-specific antigen (HpSA) was investigated by ELISA in the stool samples of patients. Resistance to tetracycline, amoxicillin, metronidazole and levofloxacin was determined with E-test method. Clarithromycin resistance was determined both by E-test and PCR-RFLP (restriction fragment length polymorphism) methods. H.pylori was detected in 29.6% (43/145) of patients with culture, 55.2% (80/145) of patients with urease test, 57% (65/114) of patients with HpSA test and 71.3% (102/143) of patients with PCR. The sensitivity and specificity of culture, PCR, HpSA and urease tests were determined as 52.4% and 100%, 96.3% and 62.3%, 80.3% and 81.4%, 86.6% and 85.7%, respectively. According to the E-test results, resistance to clarithromycin was 18.2%, to tetracycline 9.1%, to metronidazole 45.5%, to levofloxacin 18.2% and no resistance was determined to amoxicillin. Clarithromycin resistance was searched in 94 of PCR positive 102 samples, and 17 (18.1%) of them yielded clarithromycin resistance. Of them 11 (64.7%) harbored A2144G (at 2144. nucleotide), and 6 (%35.3) harbored A2143G (at 2143. nucleotide) point mutations. In our study, PCR was determined as the most sensitive method, however due to its low specificity, the results should be confirmed with at least one of the other methods. The specificity of culture method was high, but sensitivity was found to be quite low compared with other methods. The sensitivity and specificity of urease and HpSA tests were found to be similar. In conclusion, in cases which endoscopy could not be done, non-invasive, rapid and practical HpSA method can be used in diagnosis and monitorization of the treatment. In the case of treatment failure, culture should be performed for antibiotic susceptibility testing of the isolate.
幽门螺杆菌被认为是胃炎、胃和十二指肠溃疡、胃腺癌及黏膜相关淋巴组织淋巴瘤的病原体。在幽门螺杆菌感染的诊断中,可采用侵入性方法(培养、组织病理学检查、快速尿素酶试验和分子检测)和非侵入性方法(尿素呼气试验、血清学检测、粪便培养及粪便抗原/核酸检测)。为治疗和根除感染,应用了克拉霉素、阿莫西林以及甲硝唑与质子泵抑制剂或枸橼酸铋雷尼替丁的联合三联治疗方案。然而,幽门螺杆菌菌株中抗生素耐药率的不断上升降低了根除治疗的成功率。本研究的目的是调查胃窦活检标本中幽门螺杆菌的存在情况,并确定分离菌株的抗菌耐药性。本研究纳入了149例因消化不良症状入住梅尔辛大学医学院内科胃肠病诊所的患者(年龄范围:17 - 83岁;男性73例)的胃窦活检标本。通过对胃活检标本进行培养、聚合酶链反应(PCR)和尿素酶试验来检测幽门螺杆菌的存在情况,并通过酶联免疫吸附测定法(ELISA)检测患者粪便样本中的幽门螺杆菌特异性抗原(HpSA)。采用E试验法测定对四环素、阿莫西林、甲硝唑和左氧氟沙星的耐药性。采用E试验法和PCR - 限制性片段长度多态性(RFLP)方法测定克拉霉素耐药性。培养法在43/145例患者(29.6%)中检测到幽门螺杆菌,尿素酶试验在80/145例患者(55.2%)中检测到,HpSA试验在65/114例患者(57%)中检测到,PCR在102/143例患者(71.3%)中检测到。培养法、PCR、HpSA和尿素酶试验的敏感性和特异性分别确定为52.4%和100%、96.3%和62.3%、80.3%和81.4%、86.6%和85.7%。根据E试验结果,对克拉霉素的耐药率为18.2%,对四环素为9.1%,对甲硝唑为45.5%,对左氧氟沙星为18.2%,对阿莫西林未检测到耐药。在102例PCR阳性样本中的94例中检测克拉霉素耐药性,其中17例(18.1%)出现克拉霉素耐药。其中11例(64.7%)存在A2144G(第2144位核苷酸)点突变,6例(35.3%)存在A2143G(第2143位核苷酸)点突变。在我们的研究中,PCR被确定为最敏感的方法,然而由于其特异性较低,结果应至少用其他方法之一进行确认。培养法的特异性较高,但与其他方法相比敏感性相当低。尿素酶试验和HpSA试验的敏感性和特异性相似。总之,在内镜检查无法进行的情况下,非侵入性、快速且实用的HpSA方法可用于诊断和治疗监测。在治疗失败的情况下,应进行培养以对分离菌株进行抗生素敏感性检测。