Discovery of a Novel Mutation in DNA Gyrase and Changes in the Fluoroquinolone Resistance of Helicobacter pylori over a 14-Year Period: A Single Center Study in Korea
Abstract
:1. Introduction
2. Results
2.1. Patient Demographics and H. pylori Strains
2.2. Prevalence of Fluoroquinolone Resistance and Distribution of Minimum Inhibitory Concentrations (MICs)
2.3. The Mutation Analyses of DNA Gyrase Genes and the Association with MIC Levels for Fluoroquinolones
2.4. Transformation of H. pylori and Confirmation of Gyrase Mutations Related to Fluoroquinolone Resistance
3. Discussion
4. Materials and Methods
4.1. Study Population
4.2. H. pylori Culture and Isolation
4.3. PCR-Based Randomly Amplified Polymorphic DNA (RAPD) Fingerprinting of H. pylori
4.4. Determination of MICs of Fluoroquinolones
4.5. PCR Amplification and Nucleotide Sequencing
4.6. Natural Transformation of H. pylori
4.7. Statistical Methods
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Characteristics | 2005/2006 | 2017/2018 | p-Value |
---|---|---|---|
Number of patients (strains) | 143 (153) | 48 (48) | - |
Mean age ± SD (years) | 48.6 ± 14.6 | 61.9 ± 8.7 | <0.001 |
Sex (M/F) | 87/56 (60.8/39.2%) | 33/15 (68.8/31.2%) | 0.326 |
Endoscopic diagnosis | <0.001 | ||
Peptic ulcer disease | 65 (45.5%) | 15 (31.3%) | |
Gastric cancer | 3 (2.1%) | 11 (22.9%) | |
Gastric adenoma | 0 (0.0%) | 4 (8.3%) | |
MALT lymphoma | 0 (0.0%) | 1 (2.1%) | |
Gastritis | 75 (52.4%) | 17 (35.4%) |
Prevalence of Resistant Strains (Resistant Strains/Total Strains, %) | |||
---|---|---|---|
2005/2006 | 2017/2018 | p-Value | |
Levofloxacin | 19.0% (29/153) | 43.8% (21/48) | <0.001 |
Moxifloxacin | 19.0% (29/153) | 43.8% (21/48) | <0.001 |
Group | Substitution in gyrA | Substitution in gyrB | Number of Patients | MIC (µg/mL) Value | |||
---|---|---|---|---|---|---|---|
LVX | MOX | ||||||
Mean | p-Value * | Mean | p-Value * | ||||
2005/2006 (n = 29) | Asn-87→Lys | Asp-495→His | 3 (10.3%) | 4 | 0.771 | 4 | 0.823 |
Asn-87→Lys | ND | 12 (41.4%) | 3.5 | 3.3 | |||
Asp-91→Gly | ND | 12 (41.4%) | 3 | 3 | |||
ND | ND | 2 (6.9%) | 3 | 3 | |||
2017/2018 (n = 21) | Asn-87→Lys | ND | 13 (62.0%) | 10.5 | 0.652 | 10.5 | 0.652 |
Asp-91→Asn or Gly or Tyr | ND | 5 (23.8%) | 7.2 | 7.2 | |||
Ala-88→Val | ND | 1 (4.8%) | 8 | 8 | |||
Asn-87→Lys& Ala-88→Val | ND | 1 (4.8%) | 8 | 8 | |||
Gly-85→Cys | ND | 1 (4.8%) | 8 | 8 |
Isolated Strains | Amplified PCR Product Used in Transformation | Mutation in Transformed DNA | Transformation Frequency (MIC > 1 µg/mL) | Result | |
---|---|---|---|---|---|
First | Second | ||||
Negative control Positive control QA2 QA1 QA4 QA12 | gyrB gyrA gyrB gyrB gyrB gyrB | None Asn87→Lys None Asp495→His Asp495→His Asp495→His | 2.8 × 10−9 5.7 × 10−6 3.0 × 10−9 3.4 × 10−9 2.9 × 10−9 2.5 × 10−9 | 2.5 × 10−9 4.1 × 10−6 4.9 × 10−9 3.1 × 10−9 3.4 × 10−9 1.9 × 10−9 | 2.65 × 10−9 4.9 × 10−6 3.95 × 10−9 3.25 × 10−9 3.15 × 10−9 2.2 × 10−9 |
Donor Strain | MIC of Donor Strain (μg/mL) | gyrA Mutation of Donor Strain | MIC of Transformed Strains (μg/mL) | gyrA Mutation Detected in Transformed Strains | ||
---|---|---|---|---|---|---|
LVX | MOX | LVX | MOX | |||
QB1 | 8 | 8 | Gly-85→Cys | 0.5→2 | 0.25→2 | Gly-85→Cys |
QB16 | 8 | 8 | Ala-88→Val | 0.5→0.5 | 0.25→0.25 | Ala-88→Val |
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Rhie, S.Y.; Park, J.Y.; Shin, T.-S.; Kim, J.W.; Kim, B.J.; Kim, J.G. Discovery of a Novel Mutation in DNA Gyrase and Changes in the Fluoroquinolone Resistance of Helicobacter pylori over a 14-Year Period: A Single Center Study in Korea. Antibiotics 2020, 9, 287. https://doi.org/10.3390/antibiotics9060287
Rhie SY, Park JY, Shin T-S, Kim JW, Kim BJ, Kim JG. Discovery of a Novel Mutation in DNA Gyrase and Changes in the Fluoroquinolone Resistance of Helicobacter pylori over a 14-Year Period: A Single Center Study in Korea. Antibiotics. 2020; 9(6):287. https://doi.org/10.3390/antibiotics9060287
Chicago/Turabian StyleRhie, Su Yeon, Jae Yong Park, Tae-Seop Shin, Jeong Wook Kim, Beom Jin Kim, and Jae Gyu Kim. 2020. "Discovery of a Novel Mutation in DNA Gyrase and Changes in the Fluoroquinolone Resistance of Helicobacter pylori over a 14-Year Period: A Single Center Study in Korea" Antibiotics 9, no. 6: 287. https://doi.org/10.3390/antibiotics9060287