Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i>
<i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> are the predominant thermophilic species responsible for foodborne gastroenteritis worldwide. Elevated resistance to certain antibiotics was observed due to antimicrobial therapy in farm animals and humans, while reduc...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-12-01
|
| Series: | Microorganisms |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-2607/11/12/2927 |
| _version_ | 1797379981692633088 |
|---|---|
| author | Véronique Zeller-Péronnet Nancy Bretschneider Johanna Lausch Nadera Hanifi Melanie Pavlovic Michael Zarske Huong Quynh Luu Ulrich Busch Kerstin Stingl Ingrid Huber |
| author_facet | Véronique Zeller-Péronnet Nancy Bretschneider Johanna Lausch Nadera Hanifi Melanie Pavlovic Michael Zarske Huong Quynh Luu Ulrich Busch Kerstin Stingl Ingrid Huber |
| author_sort | Véronique Zeller-Péronnet |
| collection | DOAJ |
| description | <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> are the predominant thermophilic species responsible for foodborne gastroenteritis worldwide. Elevated resistance to certain antibiotics was observed due to antimicrobial therapy in farm animals and humans, while reduced antimicrobial usage partially reduced antibiotic resistance. Monitoring the antimicrobial resistance demonstrated a substantial fraction of multi-resistant isolates, indicating the necessity of reliable tools for their detection. In this study, resistance determinants in 129 German and 21 Vietnamese isolates were selected to establish a novel multiplex real-time PCR (qPCR), facilitating the simultaneous detection of four resistance determinants. These comprised <i>tet</i>(O) gene variants associated with tetracycline resistance, point mutations GyrA_T86I and GyrA_T86V associated with ciprofloxacin resistance, and the <i>erm</i>(B) gene together with the point mutation A2075G in the 23S rRNA gene, associated with erythromycin resistance. Moreover, the performance of the qPCR assay was evaluated by comparing the results of qPCR to phenotypic antimicrobial resistance profiles, obtained with standardized EUCAMP3 microdilution panel, which showed 100% similarity (inclusivity and exclusivity). Variation in measurement methods, including qPCR machines and master mixes showed robustness, essential for laboratories. The assay can be used for the rapid detection of resistance determinants, and is beneficial for monitoring the spread of antibiotic resistance in <i>C. jejuni</i> and <i>C. coli.</i> |
| first_indexed | 2024-03-08T20:30:52Z |
| format | Article |
| id | doaj.art-60fd15c4a1244f9381d420665195b0e9 |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-08T20:30:52Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-60fd15c4a1244f9381d420665195b0e92023-12-22T14:26:06ZengMDPI AGMicroorganisms2076-26072023-12-011112292710.3390/microorganisms11122927Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i>Véronique Zeller-Péronnet0Nancy Bretschneider1Johanna Lausch2Nadera Hanifi3Melanie Pavlovic4Michael Zarske5Huong Quynh Luu6Ulrich Busch7Kerstin Stingl8Ingrid Huber9Department for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyNational Reference Laboratory for Campylobacter, Department of Biological Safety, German Federal Institute for Risk Assessment (BfR), 10589 Berlin, GermanyNational Institute of Veterinary Research (NIVR), Hanoi 100000, VietnamDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, GermanyNational Reference Laboratory for Campylobacter, Department of Biological Safety, German Federal Institute for Risk Assessment (BfR), 10589 Berlin, GermanyDepartment for Food and Food Hygiene, Bavarian Health and Food Safety Authority (LGL), 85764 Oberschleissheim, Germany<i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> are the predominant thermophilic species responsible for foodborne gastroenteritis worldwide. Elevated resistance to certain antibiotics was observed due to antimicrobial therapy in farm animals and humans, while reduced antimicrobial usage partially reduced antibiotic resistance. Monitoring the antimicrobial resistance demonstrated a substantial fraction of multi-resistant isolates, indicating the necessity of reliable tools for their detection. In this study, resistance determinants in 129 German and 21 Vietnamese isolates were selected to establish a novel multiplex real-time PCR (qPCR), facilitating the simultaneous detection of four resistance determinants. These comprised <i>tet</i>(O) gene variants associated with tetracycline resistance, point mutations GyrA_T86I and GyrA_T86V associated with ciprofloxacin resistance, and the <i>erm</i>(B) gene together with the point mutation A2075G in the 23S rRNA gene, associated with erythromycin resistance. Moreover, the performance of the qPCR assay was evaluated by comparing the results of qPCR to phenotypic antimicrobial resistance profiles, obtained with standardized EUCAMP3 microdilution panel, which showed 100% similarity (inclusivity and exclusivity). Variation in measurement methods, including qPCR machines and master mixes showed robustness, essential for laboratories. The assay can be used for the rapid detection of resistance determinants, and is beneficial for monitoring the spread of antibiotic resistance in <i>C. jejuni</i> and <i>C. coli.</i>https://www.mdpi.com/2076-2607/11/12/2927food safety<i>Campylobacter</i> spp.food and clinical isolatesantimicrobial resistance determinantssusceptibility testingreal-time PCR assay |
| spellingShingle | Véronique Zeller-Péronnet Nancy Bretschneider Johanna Lausch Nadera Hanifi Melanie Pavlovic Michael Zarske Huong Quynh Luu Ulrich Busch Kerstin Stingl Ingrid Huber Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> Microorganisms food safety <i>Campylobacter</i> spp. food and clinical isolates antimicrobial resistance determinants susceptibility testing real-time PCR assay |
| title | Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> |
| title_full | Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> |
| title_fullStr | Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> |
| title_full_unstemmed | Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> |
| title_short | Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne <i>Campylobacter jejuni</i> and <i>Campylobacter coli</i> |
| title_sort | multiplex real time pcr for the detection of tetracycline ciprofloxacin and erythromycin resistance determinants from human and foodborne i campylobacter jejuni i and i campylobacter coli i |
| topic | food safety <i>Campylobacter</i> spp. food and clinical isolates antimicrobial resistance determinants susceptibility testing real-time PCR assay |
| url | https://www.mdpi.com/2076-2607/11/12/2927 |
| work_keys_str_mv | AT veroniquezellerperonnet multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT nancybretschneider multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT johannalausch multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT naderahanifi multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT melaniepavlovic multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT michaelzarske multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT huongquynhluu multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT ulrichbusch multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT kerstinstingl multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii AT ingridhuber multiplexrealtimepcrforthedetectionoftetracyclineciprofloxacinanderythromycinresistancedeterminantsfromhumanandfoodborneicampylobacterjejuniiandicampylobactercolii |