Application of 3′-mismatched reverse primer PCR compared with real-time PCR and PCR-RFLP for the rapid detection of 23S rDNA mutations associated with clarithromycin resistance in Helicobacter pylori

• Published in: International journal of antimicrobial agents 2004-04, Vol.23 (4), p.349-355
• Main Authors: Elviss, Nicola C, Lawson, Andrew J, Owen, Robert J
• Format: Article
• Language: English
• Subjects: 23S rDNA mutations; Anti-Bacterial Agents - pharmacology; Antibacterial agents; Antibiotics. Antiinfectious agents. Antiparasitic agents; Biological and medical sciences; Clarithromycin - pharmacology; Clarithromycin resistance; DNA Primers; DNA, Ribosomal - genetics; Drug Resistance, Bacterial - genetics; Helicobacter pylori; Helicobacter pylori - drug effects; Helicobacter pylori - genetics; Humans; Medical sciences; Microbial Sensitivity Tests; Mutation; Pharmacology. Drug treatments; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; RNA, Ribosomal, 23S - genetics
• ISSN: 0924-8579; 1872-7913
• DOI: 10.1016/j.ijantimicag.2003.09.015

Helicobacter pylori clarithromycin (Cla) resistance dramatically reduces efficacy of eradication therapy. In this study, 3′-mismatched reverse primer PCR (3M-PCR), real-time PCR (LightCycler), and PCR-RFLP assays were investigated to determine their sensitivity for detecting clarithromycin resistance associated with 23S rDNA mutations (A2142G, A2142C, and A2143G). For 84.8% (123/145) of isolates, the same allelic type was detected by each method although methods differed in efficiency of detecting mutations in cultures either containing mixtures of two alleles (24 isolates), or that were dual allelic variants (two isolates). The novel 3M-PCR assay format was the most sensitive, detecting all alleles at ≥0.02 ng/μl in DNA mixtures, and thus provides more precise information to guide clinical management of patients at risk of treatment failure.