Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa
Acinetobacter baumannii (A. baumannii) has developed several resistance mechanisms. The bacteria have been reported as origin of multiple outbreaks. This study aims to investigate the use of efflux pumps and quinolone resistance-associated genotypic mutations as mechanisms of resistance in A. bauman...
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| Format: | Article |
| Language: | English |
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Wiley
2021-01-01
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| Series: | International Journal of Microbiology |
| Online Access: | http://dx.doi.org/10.1155/2021/9923816 |
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| author | Noel-David Nogbou Granny M. Nkawane Khanyisa Ntshane Charles K. Wairuri Dikwata T. Phofa Kagiso K. Mokgokong Mbudzeni Ramashia Maphoshane Nchabeleng Lawrence C. Obi Andrew M. Musyoki |
| author_facet | Noel-David Nogbou Granny M. Nkawane Khanyisa Ntshane Charles K. Wairuri Dikwata T. Phofa Kagiso K. Mokgokong Mbudzeni Ramashia Maphoshane Nchabeleng Lawrence C. Obi Andrew M. Musyoki |
| author_sort | Noel-David Nogbou |
| collection | DOAJ |
| description | Acinetobacter baumannii (A. baumannii) has developed several resistance mechanisms. The bacteria have been reported as origin of multiple outbreaks. This study aims to investigate the use of efflux pumps and quinolone resistance-associated genotypic mutations as mechanisms of resistance in A. baumannii isolates at a tertiary hospital. A total number of 103 A. baumannii isolates were investigated after identification and antimicrobial susceptibility testing by VITEK2 followed by PCR amplification of blaOXA-51. Conventional PCR amplification of the AdeABC efflux pump (adeB, adeS, and adeR) and quinolone (parC and gyrA) resistance genes were performed, followed by quantitative real-time PCR of AdeABC efflux pump genes. Phenotypic evaluation of efflux pump expression was performed by determining the difference between the MIC of tigecycline before and after exposure to an efflux pump inhibitor. The Sanger sequencing method was used to sequence the parC and gyrA amplicons. A phylogenetic tree was drawn using MEGA 4.0 to evaluate evolutionary relatedness of the strains. All the collected isolates were blaOXA-51-positive. High resistance to almost all the tested antibiotics was observed. Efflux pump was found in 75% of isolates as a mechanism of resistance. The study detected parC gene mutation in 60% and gyrA gene mutation in 85%, while 37% of isolates had mutations on both genes. A minimal evolutionary distance between the isolates was reported. The use of the AdeABC efflux pump system as an active mechanism of resistance combined with point mutation mainly in gyrA was shown to contribute to broaden the resistance spectrum of A. baumannii isolates. |
| format | Article |
| id | doaj-art-5d6c70184a654b759f9872a27e2d6eb8 |
| institution | Kabale University |
| issn | 1687-918X 1687-9198 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Microbiology |
| spelling | doaj-art-5d6c70184a654b759f9872a27e2d6eb82025-02-03T07:24:04ZengWileyInternational Journal of Microbiology1687-918X1687-91982021-01-01202110.1155/2021/99238169923816Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South AfricaNoel-David Nogbou0Granny M. Nkawane1Khanyisa Ntshane2Charles K. Wairuri3Dikwata T. Phofa4Kagiso K. Mokgokong5Mbudzeni Ramashia6Maphoshane Nchabeleng7Lawrence C. Obi8Andrew M. Musyoki9Department of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaSchool of Science and Technology, Sefako Makgatho Health Sciences University, Pretoria, South AfricaDepartment of Microbiological Pathology, School of Medicine, Sefako Makgatho Health Sciences University, Pretoria, South AfricaAcinetobacter baumannii (A. baumannii) has developed several resistance mechanisms. The bacteria have been reported as origin of multiple outbreaks. This study aims to investigate the use of efflux pumps and quinolone resistance-associated genotypic mutations as mechanisms of resistance in A. baumannii isolates at a tertiary hospital. A total number of 103 A. baumannii isolates were investigated after identification and antimicrobial susceptibility testing by VITEK2 followed by PCR amplification of blaOXA-51. Conventional PCR amplification of the AdeABC efflux pump (adeB, adeS, and adeR) and quinolone (parC and gyrA) resistance genes were performed, followed by quantitative real-time PCR of AdeABC efflux pump genes. Phenotypic evaluation of efflux pump expression was performed by determining the difference between the MIC of tigecycline before and after exposure to an efflux pump inhibitor. The Sanger sequencing method was used to sequence the parC and gyrA amplicons. A phylogenetic tree was drawn using MEGA 4.0 to evaluate evolutionary relatedness of the strains. All the collected isolates were blaOXA-51-positive. High resistance to almost all the tested antibiotics was observed. Efflux pump was found in 75% of isolates as a mechanism of resistance. The study detected parC gene mutation in 60% and gyrA gene mutation in 85%, while 37% of isolates had mutations on both genes. A minimal evolutionary distance between the isolates was reported. The use of the AdeABC efflux pump system as an active mechanism of resistance combined with point mutation mainly in gyrA was shown to contribute to broaden the resistance spectrum of A. baumannii isolates.http://dx.doi.org/10.1155/2021/9923816 |
| spellingShingle | Noel-David Nogbou Granny M. Nkawane Khanyisa Ntshane Charles K. Wairuri Dikwata T. Phofa Kagiso K. Mokgokong Mbudzeni Ramashia Maphoshane Nchabeleng Lawrence C. Obi Andrew M. Musyoki Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa International Journal of Microbiology |
| title | Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa |
| title_full | Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa |
| title_fullStr | Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa |
| title_full_unstemmed | Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa |
| title_short | Efflux Pump Activity and Mutations Driving Multidrug Resistance in Acinetobacter baumannii at a Tertiary Hospital in Pretoria, South Africa |
| title_sort | efflux pump activity and mutations driving multidrug resistance in acinetobacter baumannii at a tertiary hospital in pretoria south africa |
| url | http://dx.doi.org/10.1155/2021/9923816 |
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