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

2 Citations (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

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 Kim, {Jie Hyun} and Jeong, {Su Jin} and Park, {Soon Young} and Kang, {Il Mo} and Lee, {Kyoung Hwa} and Song, {Young Goo}",
year = "2018",
month = "11",
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, 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

Y1 - 2018/11

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

C2 - 30037168

AN - SCOPUS:85057082835

VL - 12

SP - 641

EP - 647

JO - Gut and Liver

JF - Gut and Liver

SN - 1976-2283

IS - 6

ER -