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

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)641-647
Number of pages7
JournalGut and liver
Volume12
Issue number6
DOIs
Publication statusPublished - 2018 Nov 1

Fingerprint

Nucleic Acid Probes
Peptide Nucleic Acids
Clarithromycin
Helicobacter pylori
Freezing
Mutation
Point Mutation
Fluorescence
RNA
Temperature
Genes

All Science Journal Classification (ASJC) codes

  • Hepatology
  • Gastroenterology

Cite this

Jung, Da Hyun ; Kim, Jie-Hyun ; Jeong, Su Jin ; Park, Soon Young ; Kang, Il Mo ; Lee, Kyoung Hwa ; Song, Young Goo. / Peptide nucleic acid probe-based analysis as a new detection method for clarithromycin resistance in helicobacter pylori. In: Gut and liver. 2018 ; Vol. 12, No. 6. pp. 641-647.
@article{49b6348bb3714e15bc19491eb8075f23,
title = "Peptide nucleic acid probe-based analysis as a new detection method for clarithromycin resistance in helicobacter pylori",
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.",
author = "Jung, {Da Hyun} and Jie-Hyun Kim and Jeong, {Su Jin} and Park, {Soon Young} and Kang, {Il Mo} and Lee, {Kyoung Hwa} and Song, {Young Goo}",
year = "2018",
month = "11",
day = "1",
doi = "10.5009/gnl18111",
language = "English",
volume = "12",
pages = "641--647",
journal = "Gut and Liver",
issn = "1976-2283",
publisher = "Joe Bok Chung",
number = "6",

}

Peptide nucleic acid probe-based analysis as a new detection method for clarithromycin resistance in helicobacter pylori. / Jung, Da Hyun; Kim, Jie-Hyun; Jeong, Su Jin; Park, Soon Young; Kang, Il Mo; Lee, Kyoung Hwa; Song, Young Goo.

In: Gut and liver, Vol. 12, No. 6, 01.11.2018, p. 641-647.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Peptide nucleic acid probe-based analysis as a new detection method for clarithromycin resistance in helicobacter pylori

AU - Jung, Da Hyun

AU - Kim, Jie-Hyun

AU - Jeong, Su Jin

AU - Park, Soon Young

AU - Kang, Il Mo

AU - Lee, Kyoung Hwa

AU - Song, Young Goo

PY - 2018/11/1

Y1 - 2018/11/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85057082835&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85057082835&partnerID=8YFLogxK

U2 - 10.5009/gnl18111

DO - 10.5009/gnl18111

M3 - Article

VL - 12

SP - 641

EP - 647

JO - Gut and Liver

JF - Gut and Liver

SN - 1976-2283

IS - 6

ER -