Quantitative multiplex real-time polymerase chain reaction assay for the detection of Helicobacter pylori and clarithromycin resistance

• Published in: BMC microbiology Vol. 23; no. 1; pp. 155 - 8
• Main Authors: Kim, Ilsoo, Maeng, Lee-So, Kim, Joon Sung, Kim, Byung-Wook, Cheung, Dae Young, Kim, Jin Il, Park, Soo-heon
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 27.05.2023; BioMed Central; BMC
• Subjects: Accuracy; Analysis; Anti-Bacterial Agents - pharmacology; Antibiotics; Bacterial infections; Biopsy; Clarithromycin; Clarithromycin - pharmacology; DNA sequencing; Drug resistance in microorganisms; Drug Resistance, Bacterial - genetics; Endoscopy; Gastric cancer; Gene sequencing; Helicobacter infections; Helicobacter Infections - diagnosis; Helicobacter Pylori; Helicobacter pylori - genetics; Humans; Infections; Kappa coefficient; Macrolide Resistance; Molecular diagnosis; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Mutation; Oligonucleotides; Performance evaluation; Polymerase chain reaction; Prevention; Priming; Real-Time Polymerase Chain Reaction; Ribosomal RNA; Risk factors; RNA, Ribosomal, 23S - genetics; Sensitivity; Testing; Ulcers; South Korea; Macrolide Resistance; Helicobacter Pylori; Molecular diagnosis
• ISSN: 1471-2180; 1471-2180
• DOI: 10.1186/s12866-023-02868-z

Identifying clarithromycin resistance is essential for eradicating Helicobacter pylori (HP). Therefore, we evaluated the performance of Allplex™ H.pylori & ClariR Assay (Allplex™) for diagnosing and detecting clarithromycin resistance in HP. Subjects who underwent esophagogastroduodenoscopy between April 2020 and August 2021 at Incheon St. Mary's hospital were enrolled in this study. The diagnostic performances of Allplex™ and dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) were compared with sequencing as the gold standard. A total of 142 gastric biopsy samples were analyzed. Gene sequencing revealed 124 HP infections, 42 A2143G mutations, 2 A2142G mutations, one dual mutation, and no A2142C mutation. DPO-PCR showed 96.0% sensitivity and 100.0% specificity for HP detection; the corresponding rates for Allplex™ were 99.2% and 100.0%. DPO-PCR showed 88.3% sensitivity and 82.0% specificity for A2143G mutation, and Allplex™ showed 97.6% and 96.0%. The Cohen's Kappa coefficient for overall test results was 0.56 for DPO-PCR and 0.95 for Allplex™. Allplex™ showed comparable diagnostic performance with direct gene sequencing and non-inferior diagnostic performance to DPO-PCR. Further research is required to confirm whether Allplex™ is an effective diagnostic tool for the eradication of HP.