Real-Time Polymerase Chain Reaction for the Detection of Helicobacter pylori and Clarithromycin Resistance

Abstract

Background/Aims: Real-time polymerase chain reaction (RT-PCR) is a fast and simple method for the simultaneous detection of clarithromycin (CLR) resistance and Helicobacter pylori. We evaluated the effectiveness of RT-PCR compared to that of the rapid urease test (RUT) and assessed its value in verifying CLR resistance.Methods: A total of 70 specimens with confirmed H. pylori infection in culture were enrolled and analyzed in this prospective study. All specimens were subjected to RT-PCR assay using fluorescence melting peak signals to detect H. pylori infection and CLR resistances caused by either A2142G or A2143G mutations in the 23S ribosomal RNA gene (23S rRNA). The results were compared to those of RUT and antimicrobial susceptibility culturing tests to investigate the efficacy of RT-PCR.Results: Among the 70 specimens analyzed, the positivity rate was 97.1% (68/70) with RT-PCR and 82.9% (58/70) with RUT. CLR resistance (minimum inhibitory concentration >1.0 μg/mL) was confirmed in 18.6% (13/70), and fluorescence melting curve analysis showed that 84.6% (11/13) had point mutations in 23S rRNA. Ten specimens had only A2143G mutation, and one specimen contained both A2142G and A2143G mutations.Conclusions: RT-PCR assay was found to be more efficient than RUT in detecting H. pylori infection and could effectively verify CLR resistance compared to the antimicrobial susceptibility culturing test. Considering the high sensitivity and accessibility of RT-PCR method, it could be used to easily detect CLR-resistant H. pylori, thus helping clinicians select suitable treatment regimen and improve the eradication rate.

Efficacy and safety of vonoprazan–amoxicillin dual therapy for Helicobacter pylori first-line treatment: a single-center, randomized, controlled trial

“…But this method is technically demanding, time-consuming, and with a low success rate, which limits its clinical use. 34 Clarithromycin inhibits bacterial growth by targeting the 50S subunits of the ribosome. The 50S subunit of H. pylori is encoded by 23S rRNA.…”

“…Of these, the most common point mutations are A2146C, A2146G, and A2147G. Detection of the above three mutations using molecular biological approaches can identify about 85% of clarithromycin-resistant strains, 34 , 35 which can meet clinical needs. In our study, we detected H. pylori 23S rRNA mutations associated with clarithromycin resistance (A2142-43G, A2142C) using real-time PCR technique and analyzed the effect of clarithromycin resistance on the eradication rates of VA dual therapy.…”

Optimizing Helicobacter pylori Treatment: An Updated Review of Empirical and Susceptibility Test-Based Treatments

“…78 Another study compared the diagnostic accuracy of real-time PCR against MIC measurements by culture test, and it showed a high concordance rate; 84.6% of subjects diagnosed with clarithromycin-resistant infections by the culture method also had point mutations in 23S rRNA. 79 DNA gyrase, which is used to break the DNA double helix structure during DNA replication, is required for H. pylori to proliferate. DNA gyrase exists as a tetramer formed by the subunits encoded by the gyrA and gyrB genes.…”

Bismuth-Based Quadruple Therapy as First-Line Treatment for Clarithromycin-Resistant Helicobacter pylori Infection: A Prospective Randomized Comparison of 7- and 14-Day Treatment Regimens

“…2 The RT-PCR assay detects point mutations in the 23S rRNA gene of HP, and the A2142G and A2143G point mutations have been reported to be found in clarithromycinresistant HP strains. 3,4 HP eradication with identification of clarithromycin resistance has shown high success rates. [5][6][7][8] Bismuth-based quadruple therapy (BQT), consisting of a PPI, bismuth subcitrate, metronidazole, and tetracycline, is one of therapeutic options for first-line treatment where local clarithromycin resistance rates are over 15% and for clarithromycin-resistant HP infections.…”