Peptide Nucleic Acid Probe-Based Analysis as a New Detection Method for Clarithromycin Resistance in Helicobacter pylori.
- Author:
Da Hyun JUNG
1
;
Jie Hyun KIM
;
Su Jin JEONG
;
Soon Young PARK
;
Il Mo KANG
;
Kyoung Hwa LEE
;
Young Goo SONG
Author Information
- Publication Type:Original Article
- Keywords: Helicobacter pylori; Clarithromycin; Melting array; Peptide nucleic acids
- MeSH: Clarithromycin*; Fluorescence; Freezing; Helicobacter pylori*; Helicobacter*; Humans; Methods*; Peptide Nucleic Acids; Point Mutation; RNA
- From:Gut and Liver 2018;12(6):641-647
- CountryRepublic of Korea
- Language:English
- Abstract: BACKGROUND/AIMS: Helicobacter pylori eradication rates are decreasing because of increases in clarithromycin resistance. Thus, finding an easy and accurate method of detecting clarithromycin resistance is important. METHODS: We evaluated 70 H. pylori isolates from Korean patients. Dual-labeled peptide nucleic acid (PNA) probes were designed to detect resistance associated with point mutations in 23S ribosomal ribonucleic acid gene domain V (A2142G, A2143G, and T2182C). Data were analyzed by probe-based fluorescence melting curve analysis based on probe-target dissociation temperatures and compared with Sanger sequencing. RESULTS: Among 70 H. pylori isolates, 0, 16, and 58 isolates contained A2142G, A2143G, and T2182C mutations, respectively. PNA probe-based analysis exhibited 100.0% positive predictive values for A2142G and A2143G and a 98.3% positive predictive value for T2182C. PNA probe-based analysis results correlated with 98.6% of Sanger sequencing results (κ-value=0.990; standard error, 0.010). CONCLUSIONS: H. pylori clarithromycin resistance can be easily and accurately assessed by dual-labeled PNA probe-based melting curve analysis if probes are used based on the appropriate resistance-related mutations. This method is fast, simple, accurate, and adaptable for clinical samples. It may help clinicians choose a precise eradication regimen.
