DETERMINATION OF ERYTHROMYCIN AND PENICILLIN RESISTANCE GENES IN STREPTOCOCCUS PNEUMONIAE FROM PATIENTS ATTENDING SELECTED HOSPITALS IN KADUNA STATE, NIGERIA

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Date
2019-06
Authors
LUSA, Haruna Yadock
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Abstract
Streptococcus pneumoniae (Pneumococcus) is both an aggressive pathogen and a normal flora of the human microbiome with an array of over 90 serotypes.Penicillin-resistant S. pneumoniae (PRP) and Erythromycin resistant Pneumococci (ERP) has emerged as a worldwide problem. S. pneumoniae develops penicillin resistance through recombination with other PRP strains or closely related streptococcal species that alter its penicillin binding protein (PbP) genes pbp1a, pbp2b, and pbp2x. These altered PbP gene sequences of PRP differ greatly within the PRP subset and from the sequences of penicillin-susceptible S.pneumoniae (PSSP),whileresistanceto macrolides in S.pneumoniae arises primarily due to macrolide resistant Genes (MRGs) including; erythromycin ribose methyltransferase encoded by ermB or an efflux system leading to selective efflux of 14 and15 membered macrolides encoded by mef(A).Erythromycin ribose methyltransferase (ermB) typically confers high level resistance to macrolides, lincosamides and streptogramin B (MLSB phenotype) whereas mef A confers low le vel resistance to macrolides. The study aimed to detect streptococcus pneumoniae strains with erythromycin and penicillin resistance genes. A facility based cross sectional study with 450 patients attending selected hospitals in Kaduna State. The enrolled patients were selected by systematic random sampling presenting with clinically suspected respiratory tract infections (RTIs). Two hundred and thirty-seven (52.7%) of the subjects were males and 213(47.3%) were females aged between 10- 70 years with symptoms of respiratory tract infections. Two hundred and seven (46.0%) of the samples yielded positive growth of which 41(9.1%) were Streptococcus pneumoniae isolated in 5% blood agar in the presence of 5% CO2,,morphology, identified with bile solubility, optochin and antibiotic susceptibilities. MIC determination was done with E-test for penicillin and erythromycin while molecular determination was done for serotypes macrolide and penicillin resistance genes by PCR. Incidence of 20.5% Penicillin resistant pneumococci (PRP) and 29.4% Erythromycin resistant pneumococci (ERP) were established among the elderly patients age ≥ 50 years. Economic status of the patients had a significant relationship to Penicillin and Erythromycin resistance (P=0.0004**) determined by disc diffusion technique. Thirty-one (75.6%) of the Pneumococcal isolates (n=41) were penicillin non-susceptible pneumococci (PNSP)[MICs 1.0-15.5μg/mL; MIC50=8.0μg/mL) and 31(75.6%) were erythromycin non-susceptible pneumococci(ENSP) [MICs 0.12-6.0μg/mL;MIC50=0.90μg/mL]determined by E-Test methodology. Four pneumococcal serotypes were detected in the study area: 3(33.3%), 13(33.3%), 20(16.7%), 23F (16.7%) by conventional PCR serotyping strategy. PCR assay and Agar-gel electrophoresis for the simultaneous detection of lyt A, Penicillin resistance genes (PRGs: PbP1a, 2b, 2x) and Macrolide resistance genes (MRGs: ermB and Mef A) was carried out. The results show 3 of the pneumococcal strains harbor mutations in PbP1a and PbP 2x; six of the strains express ermB and mefA genes. This study shows that pneumococcal drug resistance in the area is predominantly mediated by ermB and mefA genes with serotypes 3 and 13 having the highest frequency among the isolates suggesting the use of more specific gene targets may in the near future solve the problem of inaccurate identification of pneumococcal infections of the lower respiratory tract. The finding suggests the use of more specific gene targets in the near future may solve the problem of inaccurate identification of pneumococcal infections of the lower respiratory tract. This study also highlights the danger in empirical use of these groups of antibiotics. Our facilities must be enforced to put in place policies that will guarantee protection and safety of patients, their relatives and heath personnel.
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A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES AHMADU BELLO UNIVERSITY IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF DOCTOR OF PHYLOSOPHY DEGREE (Ph.D.) IN MEDICAL MICROBIOLOGY DEPARTMENT OF MICROBIOLOGY, FACULTY OF LIFE SCIENCES, AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA
Keywords
ERYTHROMYCIN,, PENICILLIN RESISTANCE GENES,, STREPTOCOCCUS PNEUMONIAE,, PATIENTS,, KADUNA STATE,, NIGERIA.
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