Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis
Introduction. Treatment of gonorrhoea infection is limited by the increasing prevalence of multidrug-resistant strains. Cost-effective molecular diagnostic tests can guide effective antimicrobial stewardship. The aim of this study was to correlate mRNA expression levels in Neisseria gonorrhoeae anti...
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| Format: | Article |
| Language: | English |
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Wiley
2022-01-01
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| Series: | International Journal of Microbiology |
| Online Access: | http://dx.doi.org/10.1155/2022/7318325 |
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| author | Nireshni Mitchev Ravesh Singh Veron Ramsuran Arshad Ismail Mushal Allam Stanford Kwenda Florah Mnyameni Nigel Garrett Khine Swe Swe-Han Abraham J. Niehaus Koleka P. Mlisana |
| author_facet | Nireshni Mitchev Ravesh Singh Veron Ramsuran Arshad Ismail Mushal Allam Stanford Kwenda Florah Mnyameni Nigel Garrett Khine Swe Swe-Han Abraham J. Niehaus Koleka P. Mlisana |
| author_sort | Nireshni Mitchev |
| collection | DOAJ |
| description | Introduction. Treatment of gonorrhoea infection is limited by the increasing prevalence of multidrug-resistant strains. Cost-effective molecular diagnostic tests can guide effective antimicrobial stewardship. The aim of this study was to correlate mRNA expression levels in Neisseria gonorrhoeae antibiotic target genes and efflux pump genes to antibiotic resistance in our population. Methods. This study investigated the expression profile of antibiotic resistance-associated genes (penA, ponA, pilQ, mtrR, mtrA, mtrF, gyrA, parC, parE, rpsJ, 16S rRNA, and 23S rRNA) and efflux pump genes (macAB, norM, and mtrCDE), by quantitative real-time PCR, in clinical isolates from KwaZulu-Natal, South Africa. Whole-genome sequencing was used to determine the presence or absence of mutations. Results. N. gonorrhoeae isolates, from female and male patients presenting for care at clinics in KwaZulu-Natal, South Africa, were analysed. As determined by binomial regression and ROC analysis, the most significant (p≤0.05) markers for resistance prediction in this population, and their cutoff values, were determined to be mtrC (p=0.024; cutoff <0.089), gyrA (p=0.027; cutoff <0.0518), parE (p=0.036; cutoff <0.0033), rpsJ (p=0.047; cutoff <0.0012), and 23S rRNA (p=0.042; cutoff >7.754). Conclusion. Antimicrobial stewardship includes exploring options to conserve currently available drugs for gonorrhoea treatment. There is the potential to predict an isolate as either susceptible or nonsusceptible based on the mRNA expression level of specific candidate markers, to inform patient management. This real-time qPCR approach, with few targets, can be further investigated for use as a potentially cost-effective diagnostic tool to detect resistance. |
| format | Article |
| id | doaj-art-c491591fb5c743a18c17b0576d543c1c |
| institution | Kabale University |
| issn | 1687-9198 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Microbiology |
| spelling | doaj-art-c491591fb5c743a18c17b0576d543c1c2025-02-03T01:07:56ZengWileyInternational Journal of Microbiology1687-91982022-01-01202210.1155/2022/7318325Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression AnalysisNireshni Mitchev0Ravesh Singh1Veron Ramsuran2Arshad Ismail3Mushal Allam4Stanford Kwenda5Florah Mnyameni6Nigel Garrett7Khine Swe Swe-Han8Abraham J. Niehaus9Koleka P. Mlisana10School of Laboratory Medicine and Medical SciencesSchool of Laboratory Medicine and Medical SciencesSchool of Laboratory Medicine and Medical SciencesSequencing Core FacilitySequencing Core FacilitySequencing Core FacilitySequencing Core FacilityCentre for the AIDS Programme of Research in South AfricaSchool of Laboratory Medicine and Medical SciencesSchool of Laboratory Medicine and Medical SciencesSchool of Laboratory Medicine and Medical SciencesIntroduction. Treatment of gonorrhoea infection is limited by the increasing prevalence of multidrug-resistant strains. Cost-effective molecular diagnostic tests can guide effective antimicrobial stewardship. The aim of this study was to correlate mRNA expression levels in Neisseria gonorrhoeae antibiotic target genes and efflux pump genes to antibiotic resistance in our population. Methods. This study investigated the expression profile of antibiotic resistance-associated genes (penA, ponA, pilQ, mtrR, mtrA, mtrF, gyrA, parC, parE, rpsJ, 16S rRNA, and 23S rRNA) and efflux pump genes (macAB, norM, and mtrCDE), by quantitative real-time PCR, in clinical isolates from KwaZulu-Natal, South Africa. Whole-genome sequencing was used to determine the presence or absence of mutations. Results. N. gonorrhoeae isolates, from female and male patients presenting for care at clinics in KwaZulu-Natal, South Africa, were analysed. As determined by binomial regression and ROC analysis, the most significant (p≤0.05) markers for resistance prediction in this population, and their cutoff values, were determined to be mtrC (p=0.024; cutoff <0.089), gyrA (p=0.027; cutoff <0.0518), parE (p=0.036; cutoff <0.0033), rpsJ (p=0.047; cutoff <0.0012), and 23S rRNA (p=0.042; cutoff >7.754). Conclusion. Antimicrobial stewardship includes exploring options to conserve currently available drugs for gonorrhoea treatment. There is the potential to predict an isolate as either susceptible or nonsusceptible based on the mRNA expression level of specific candidate markers, to inform patient management. This real-time qPCR approach, with few targets, can be further investigated for use as a potentially cost-effective diagnostic tool to detect resistance.http://dx.doi.org/10.1155/2022/7318325 |
| spellingShingle | Nireshni Mitchev Ravesh Singh Veron Ramsuran Arshad Ismail Mushal Allam Stanford Kwenda Florah Mnyameni Nigel Garrett Khine Swe Swe-Han Abraham J. Niehaus Koleka P. Mlisana Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis International Journal of Microbiology |
| title | Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis |
| title_full | Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis |
| title_fullStr | Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis |
| title_full_unstemmed | Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis |
| title_short | Assessment of Antibiotic Resistance and Efflux Pump Gene Expression in Neisseria Gonorrhoeae Isolates from South Africa by Quantitative Real-Time PCR and Regression Analysis |
| title_sort | assessment of antibiotic resistance and efflux pump gene expression in neisseria gonorrhoeae isolates from south africa by quantitative real time pcr and regression analysis |
| url | http://dx.doi.org/10.1155/2022/7318325 |
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