Department of Pediatrics, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga-shi, Saga, 849-8501, Japan.
Division of Gastroenterology, Department of Internal Medicine, Imamura Hospital, Tosu, Japan.
BMC Gastroenterol. 2020 Nov 23;20(1):397. doi: 10.1186/s12876-020-01549-9.
The resistance rate of Helicobacter pylori to clarithromycin (CAM) is high among infected children in Japan. Therefore, a new method for detecting CAM-resistant H. pylori using a minimally invasive technique is strongly desired. We aimed to investigate the clinical usefulness of our newly developed nested polymerase chain reaction-quenching probe (Nested PCR-QP) method using stool specimens.
We first evaluated our method using a residual solution of the H. pylori stool antigen test for adolescents. Then, we evaluated our method using culture testing for adults.
Among 57 middle school students with H. pylori, the Nested PCR-QP test results of 53 (90.3%) were able to be analyzed. A total of 28 students had CAM resistance mutations. We found a genetic mutation in 28 students and no mutation in 23 students, and these results were consistent with those of PCR-direct sequencing. In the 23 adults who were diagnosed with H. pylori infection using the rapid urease test and culture testing, we were able to use Nested PCR-QP for analyzing 21 adults who tested positive in the stool H. pylori antigen test. The results obtained for all 21 adults were consistent with those obtained via the drug susceptibility test.
Our novel method could be useful for non-invasively detecting CAM resistance mutations in H. pylori. This may help select a drug to reduce eradication failure rates against H. pylori. Trial registration This study was registered with the University Hospital Medical Information Network Clinical Trials Registry (no. UMIN000030632, https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000034977 ) on 29 December 2017.
在日本，感染儿童的幽门螺杆菌对克拉霉素（CAM）的耐药率很高。因此，强烈需要一种使用微创技术检测 CAM 耐药幽门螺杆菌的新方法。我们旨在使用粪便标本研究我们新开发的嵌套聚合酶链反应-淬灭探针（Nested PCR-QP）方法的临床实用性。
我们首先使用青少年幽门螺杆菌粪便抗原检测的残留溶液评估我们的方法。然后，我们使用成人培养检测评估我们的方法。
在 57 名中学生中，53 名（90.3%）的 Nested PCR-QP 检测结果可进行分析。共有 28 名学生有 CAM 耐药突变。我们在 28 名学生中发现了遗传突变，在 23 名学生中没有发现突变，这些结果与 PCR-直接测序的结果一致。在通过快速尿素酶试验和培养试验诊断为幽门螺杆菌感染的 23 名成年人中，我们能够使用 Nested PCR-QP 分析 21 名粪便幽门螺杆菌抗原检测呈阳性的成年人。所有 21 名成年人的结果均与药敏试验结果一致。
我们的新方法可用于非侵入性地检测幽门螺杆菌中的 CAM 耐药突变。这可能有助于选择药物来降低幽门螺杆菌根除失败率。
本研究于 2017 年 12 月 29 日在大学医院医疗信息网络临床试验注册中心（注册号 UMIN000030632，https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000034977）注册。
