High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens
Background. Antimicrobial resistance is limiting treatment options for Neisseria gonorrhoeae infections. To aid or replace culture and the syndromic management approach, molecular assays are required for antimicrobial susceptibility testing to guide appropriate and rapid treatment. Objective. We aim...
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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/10.1155/2022/9094328 |
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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 | Background. Antimicrobial resistance is limiting treatment options for Neisseria gonorrhoeae infections. To aid or replace culture and the syndromic management approach, molecular assays are required for antimicrobial susceptibility testing to guide appropriate and rapid treatment. Objective. We aimed to detect single-nucleotide polymorphisms and plasmids associated with antimicrobial resistance from N. gonorrhoeae isolates from a clinic population in South Africa, using real-time PCR as a rapid test for AMR detection. Methods. N. gonorrhoeae isolates, from female and male patients presenting for care at a sexually transmitted infections clinic in Durban, South Africa, were analysed using phenotypic and genotypic methods for identification and antibiotic susceptibility testing (AST). Real-time PCR and high-resolution melting analysis were used to detect porA pseudogene (species-specific marker) and resistance-associated targets. Whole-genome sequencing was used as the gold standard for the presence of point mutations. Results. The real-time porA pseudogene assay identified all N. gonorrhoeae-positive isolates and specimens. Concordance between molecular detection (real-time PCR and HRM) and resistance phenotype was ≥92% for blaTEM (HLR penicillin), rpsJ_V57M (tetracycline), tetM (tetracycline), and gyrA_S91F (ciprofloxacin). Resistance determinants 16SrRNA_C1192U (spectinomycin), mtrR_G45D (azithromycin), and penA_D545S, penA_mosaic (cefixime/ceftriaxone) correlated with the WHO control isolates. Conclusions. Eight resistance-associated targets correlated with phenotypic culture results. The porA pseudogene reliably detected N. gonorrhoeae. Larger cohorts are required to validate the utility of these targets as a convenient culture-free diagnostic tool, to guide STI management in a South African population. |
| format | Article |
| id | doaj-art-6918c0c8599c42c1ba362389b56a8126 |
| 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-6918c0c8599c42c1ba362389b56a81262025-02-03T06:11:19ZengWileyInternational Journal of Microbiology1687-91982022-01-01202210.1155/2022/9094328High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and SpecimensNireshni 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 SciencesBackground. Antimicrobial resistance is limiting treatment options for Neisseria gonorrhoeae infections. To aid or replace culture and the syndromic management approach, molecular assays are required for antimicrobial susceptibility testing to guide appropriate and rapid treatment. Objective. We aimed to detect single-nucleotide polymorphisms and plasmids associated with antimicrobial resistance from N. gonorrhoeae isolates from a clinic population in South Africa, using real-time PCR as a rapid test for AMR detection. Methods. N. gonorrhoeae isolates, from female and male patients presenting for care at a sexually transmitted infections clinic in Durban, South Africa, were analysed using phenotypic and genotypic methods for identification and antibiotic susceptibility testing (AST). Real-time PCR and high-resolution melting analysis were used to detect porA pseudogene (species-specific marker) and resistance-associated targets. Whole-genome sequencing was used as the gold standard for the presence of point mutations. Results. The real-time porA pseudogene assay identified all N. gonorrhoeae-positive isolates and specimens. Concordance between molecular detection (real-time PCR and HRM) and resistance phenotype was ≥92% for blaTEM (HLR penicillin), rpsJ_V57M (tetracycline), tetM (tetracycline), and gyrA_S91F (ciprofloxacin). Resistance determinants 16SrRNA_C1192U (spectinomycin), mtrR_G45D (azithromycin), and penA_D545S, penA_mosaic (cefixime/ceftriaxone) correlated with the WHO control isolates. Conclusions. Eight resistance-associated targets correlated with phenotypic culture results. The porA pseudogene reliably detected N. gonorrhoeae. Larger cohorts are required to validate the utility of these targets as a convenient culture-free diagnostic tool, to guide STI management in a South African population.http://dx.doi.org/10.1155/2022/9094328 |
| 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 High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens International Journal of Microbiology |
| title | High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens |
| title_full | High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens |
| title_fullStr | High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens |
| title_full_unstemmed | High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens |
| title_short | High-Resolution Melting Analysis to Detect Antimicrobial Resistance Determinants in South African Neisseria gonorrhoeae Clinical Isolates and Specimens |
| title_sort | high resolution melting analysis to detect antimicrobial resistance determinants in south african neisseria gonorrhoeae clinical isolates and specimens |
| url | http://dx.doi.org/10.1155/2022/9094328 |
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