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Peptide Nucleic Acid Probe-Based Analysis as a New Detection Method for Clarithromycin Resistance in Helicobacter pylori

 Da Hyun Jung  ;  Jie-Hyun Kim  ;  Su Jin Jeong  ;  Soon Young Park  ;  Il-Mo Kang  ;  Kyoung Hwa Lee  ;  Young Goo Song 
 Gut and Liver, Vol.12(6) : 641-647, 2018 
Journal Title
 Gut and Liver 
Issue Date
Clarithromycin ; Helicobacter pylori ; Melting array ; Peptide nucleic acids
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.
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1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
김지현(Kim, Jie-Hyun) ORCID logo https://orcid.org/0000-0002-9198-3326
송영구(Song, Young Goo) ORCID logo https://orcid.org/0000-0002-0733-4156
이경화(Lee, Kyoung Hwa) ORCID logo https://orcid.org/0000-0003-0033-1398
정다현(Jung, Da Hyun)
정수진(Jeong, Su Jin) ORCID logo https://orcid.org/0000-0003-4025-4542
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