OJO

DETECTION OF RESISTANT AND VIRULENCE GENES ON MULTIPLE ANTIBIOTIC RESISTANT STAPHYLOCOCCUS AUREUS FROM CLINICAL WOUNDS AND BURNS PATIENTS USING QUANTITATIVE POLYMERASE CHAIN REACTION (QPCR)

*1Ojo, S.K.S., 2Esumeh, F.I., 1Sunmonu, G.T.

*1Department of Microbiology, Federal University Oye-Ekiti, Ekiti State, Nigeria; 2Department of Microbiology, Ambrose Alli University, Ekpoma, Edo State, Nigeria.

*1Corresponding author/Presenter: stephen.ojo@fuoye.edu.ng; +2348067966393

Abstract

Aim: To detect the genes (resistant and virulence) associated with multiple antibiotic resistant Staphylococcus aureus isolated from wounds and burns patients.

Background: S. aureus is one of the major causes of nosocomial infections and are most profound in community in previously healthy individuals and are associated with increased rates of illness and death.

Methods: Two hundred clinical wound and burn samples were obtained from tertiary health care facilities for S. aureus isolation, which was identified and characterized using standard microbiological procedures. Methicillin resistance was determined using β-lactamase assay and oxacillin disk (Oxoid) susceptibility test. Quantification of the S. aureus strains was performed using qPCR assay with primers and probes designed for antibiotic resistance genes (mecA) and enterotoxin genes (sea). Agarose gel electrophoresis was carried out on the qPCR products using 1.5 % agarose gel with a standard DNA ladder (100 bp), visualized under UV transilluminator and the image taken using digital camera.

Results: 44 (22 %) S. aureus were isolated and characterized with 36 (82 %) of the 44 S. aureus strains producing β-lactamase enzyme and were resistant to oxacillin (MRSA) while 8 (18%) of 44    S. aureus strains do not produce β-lactamase enzyme and were sensitive to oxacillin (MSSA). The    β-lactamase and non-β-lactamase isolates were resistant to other antibiotics. The quantification of PCR products indicated that sea genes (virulence enterotoxin factor) were detected from the antibiotic resistant staphylococci ranging from 0 – 13551.84 nmoles while the quantification of mecA genes detected ranged from 0 – 2601.76 nmoles. The agarose gel electrophoresis of the PCR products of mecA and sea genes showed amplicon size of 657 bp for mecA and 526 bp for sea genes after amplification of the antibiotic resistant S. aureus strains.

Conclusion: Owing to the detection of interested genes from the MSSA isolates, there is the need for urgent clinical and pharmaceutical attention.

 

Name of Presenter: OJO, S.K.S (PhD)

Affiliation of Presenter: Department of Microbiology, Federal University Oye-Ekiti

P.M.B. 373, Oye-Ekiti, Ekiti State, Nigeria

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