Multidrug resistant (MDR) extended-spectrum β-lactamase (ESBL)-producing and carbapenem-resistant Gram-negative bacteria have become an international health issue limiting treatment options. The objective of this study was to determine the prevalence of carbapenem-susceptible (CS) and carbapenem-resistant (CR) Enterobacteriaceae, Acinetobacter baumannii and Pseudomonas aeruginosa and investigate clinical isolates for their resistant genes/ determinants. A total of 98 bacterial isolates (59 CS and 39 CR) were collected between February 2012 and February 2013 at NHLS, after being recovered from various clinical specimens from PE hospital complex. Antimicrobial susceptibility testing was performed using the VITEK 2® system, E-test and microbroth dilution method. PCR and DNA sequencing were used to investigate: (i) ESBLs: CTX-M, TEM, SHV and OXA-1; (ii) plasmidmediated quinolone resistance (PMQR) genes: qnrA, qnrB, qnrC, qnrD, qnrS, qepA and aac(6’)-lb-cr; (iii) Escherichia coli sequence type 131 (ST131); (iv) carbapenemases: NDM, VIM, IMP, KPC, BIC, SME, IMI, NMC-A, GES, OXA-23, OXA-24, OXA-48, OXA-51 and OXA-58; and (v) insertion sequence ISAba1 upstream blaOXA-23/-24/-51/-58 genes. Porin loss in CR isolates was determined by SDSPAGE while genetic relatedness between E. coli ST131 isolates was determined by pulsed-field gel electrophoresis (PFGE). MDR ESBL-producing Enterobacteriaceae (mainly K. pneumoniae) and CR A. baumannii isolates were recovered from neonatal/ infant specimens. The majority of CS and CR isolates were MDR, possessing multiple ESBL genotypes (CTX-M, TEM, SHV and OXA-1). ESBL variants identified included: CTX-M-1, CTX-M-3, CTX-M-15, CTX-M-22, CTX-M-9, CTX-M-14, TEM-1, SHV-1, SHV-11 and OXA-1. PMQRs identified included: aac(6’)- lb-cr, qnrB1, qnrB2, qnrB13 and qnrS1. Twelve of 21 (57.1 percent) E. coli isolates belonged to the ST131 clonal complex and were genetically diverse, mainly producing CTX-M-15. Carbapenem resistance mechanisms identified included: (i) OXA-23 preceded by ISAba1 in 10 A. baumannii and 2 P. aeruginosa isolates; (ii) IMI-2 carbapenemase in an E. asburiae isolate; and (iii) combination of porin loss and ESBL production in 1 K. pneumoniae and 1 E. coli isolate. This is the first report in South Africa describing the occurrence of CTX-M-9, CTX-M-22 and IMI-2 among Enterobacteriaceae; CTX-M-15 in A. baumannii; and OXA-23 in combination with OXA-51 in P. aeruginosa. However, resistance determinants could not be detected for 24 carbapenem-resistant isolates which requires further investigation.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:nmmu/vital:10353 |
| Date | January 2014 |
| Creators | Gqunta, Kwanele |
| Publisher | Nelson Mandela Metropolitan University, Faculty of Science |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis, Masters, MSc |
| Format | xvi, 191 leaves, pdf |
| Rights | Nelson Mandela Metropolitan University |
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