DETECTION AND PREVALENCE OF EFFLUX PUMP-MEDIATED DRUG RESISTANCE IN CLINICAL ISOLATES OF MULTIDRUG-RESISTANT GRAM-NEGATIVE BACTERIA FROM NORTH KERALA, INDIA

Authors

  • Manju Suresh Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635
  • Nithya N. Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635
  • Jayasree P. R. School of Health Sciences University of Calicut, Malappuram-673635
  • Manish Kumar P. R. Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

Abstract

Objectives: The present study was carried out to detect the prevalence of efflux pump-mediated drug resistance in clinical isolates of multidrugresistant
(MDR) Gram-negative
bacteria
isolated
from
North Kerala.
Methods: Clinical isolates (n = 123) of MDR Gram-negative bacteria were collected from various clinical laboratories in North Kerala, and their effluxmediated
drug resistance
was
detected
by
two
simple phenotypic assays
-
ethidium
bromide
(EB)-agar
cartwheel
method and
efflux pump inhibitor

(EPI)-based
microplate
assay,
employing
phenylalanine-arginine
β-naphthylamide
as inhibitor.
Results: The 123 Gram-negative MDR strains tested comprised Escherichia coli, Pseudomonas aeruginosa, Acinetobacter spp., and Klebsiella spp. The
EB-agar cartwheel method of screening revealed efflux activity in 20% (n=25) of the strains with representatives from all 4 genera. The efflux activity
was revealed at a minimum concentration of EB at 1 mg/l. P. aeruginosa strains showed the highest activity, many folds higher up to a concentration
of 2.5 mg/l. The confirmatory EPI-based microplate assay showed efflux activity only in 15% (n=18) strains with 6% (n=7) active against more than
one antibiotic. Efflux pump-mediated drug resistance was found to be most prevalent in P. aeruginosa (34.8%, n=8 out of 23), followed by that in E. coli
(18.6%, n=8 out of 43), Acinetobacter spp. (9%, n=1out of 11), and Klebsiella spp. (2%, n=1 out of 46).
Conclusion: This study reports on the emergence of efflux pump-based multidrug-resistance in North Kerala. Our results showed that 15% of drug
resistance in Gram-negative MDR strains is attributable to efflux-related mechanisms, thereby emphasizing the need for inclusion of efflux-related
tests in the diagnostic regimen for MDR clinical bacteria.
Keywords: Gram-negative bacteria, Multidrug-resistance, Efflux pumps, Ethidium bromide, Efflux pump-inhibitor.

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Author Biographies

Manju Suresh, Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

DST- INSPIRE Research Fellow

Nithya N., Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

ICMR- Research Fellow

Jayasree P. R., School of Health Sciences University of Calicut, Malappuram-673635

School of Health Sciences
University of Calicut, Malappuram-673635

Assistant Professor

Manish Kumar P. R., Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

Department of Biotechnology University of Calicut Malappuram District Kerala State - 673 635

Associate Professor, Former Head & Co-ordinator

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Published

01-05-2016

How to Cite

Suresh, M., N. N., J. P. R., and M. K. P. R. “DETECTION AND PREVALENCE OF EFFLUX PUMP-MEDIATED DRUG RESISTANCE IN CLINICAL ISOLATES OF MULTIDRUG-RESISTANT GRAM-NEGATIVE BACTERIA FROM NORTH KERALA, INDIA”. Asian Journal of Pharmaceutical and Clinical Research, vol. 9, no. 3, May 2016, pp. 324-7, https://mail.innovareacademics.in/journals/index.php/ajpcr/article/view/11412.

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