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Updated: Nov 29, 2025
Detection of Helicobacter pylori Infection and Antibiotic Resistance via Stool Quantitative Polymerase Chain Reaction Analysis
Published on: May 16, 2025
Assessment of a novel method to detect clarithromycin-resistant Helicobacter pylori using a stool antigen test
Toshihiko Kakiuchi1, Kazutoshi Hashiguchi2, Ichiro Imamura2
1Department of Pediatrics, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga-shi, Saga, 849-8501, Japan. kakiucht@cc.saga-u.ac.jp.
Insights
A new nested polymerase chain reaction-quenching probe (Nested PCR-QP) method effectively detects clarithromycin (CAM)-resistant Helicobacter pylori in children and adults using stool samples. This minimally invasive approach aids in selecting appropriate treatment to reduce eradication failure rates.
Area of Science:
- Microbiology
- Molecular Diagnostics
- Clinical Medicine
Background:
- High rates of Helicobacter pylori resistance to clarithromycin (CAM) in Japanese children necessitate novel detection methods.
- Minimally invasive techniques for identifying CAM-resistant H. pylori are crucial for effective treatment.
Purpose of the Study:
- To evaluate the clinical utility of a newly developed nested polymerase chain reaction-quenching probe (Nested PCR-QP) method.
- To assess the feasibility of using stool specimens for detecting CAM resistance mutations in H. pylori.
Main Methods:
- The Nested PCR-QP method was initially validated using residual solutions from H. pylori stool antigen tests in adolescents.
- Subsequently, the method was evaluated in adults using culture testing for H. pylori confirmation.
Main Results:
- The Nested PCR-QP method successfully analyzed stool samples from 90.3% of 57 H. pylori-infected middle school students.
- CAM resistance mutations were identified in 28 students, with results consistent with PCR-direct sequencing.
- In adults, the Nested PCR-QP method showed consistent results with drug susceptibility testing for detecting resistance mutations.
Conclusions:
- The novel Nested PCR-QP method offers a non-invasive approach for detecting CAM resistance mutations in H. pylori.
- This diagnostic tool has the potential to improve treatment selection and reduce H. pylori eradication failure rates.
Background:
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.
Methods:
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.
Results:
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.
Conclusions:
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.
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