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Detection of AdeABC efflux pump genes in tetracycline-resistant Acinetobacter baumannii isolates from burn and ventilator-associated pneumonia patients.

Beheshti M, Talebi M, Ardebili A, Bahador A, Lari AR - J Pharm Bioallied Sci (2014)

Bottom Line: All tetracycline resistant isolates have AdeABC in PCR assay.The reduction of tetracycline MICs by using 50 μg/ml CCCP were as follows: in 18 isolates 2-4 fold reduction in MICs, 26 isolates showed 8 fold reduction,1 isolate showed 16 fold, 1 isolate showed 32 fold and the remaining 1 isolate showed 128 fold reduction in MICs.The results showed significant correlation between tetracycline resistance and AdeABC efflux pump genes in resistant A. baumannii isolates.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Iran University of Medical Sciences, Tehran, Iran ; Department of Microbiology, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Purpose: Acinetobacter baumannii is the most prevalent nosocomial pathogen which have been emerged in the past three decades worldwide. The aim of this study was to assess the distribution of the AdeABC efflux pump genes, associated with tetracycline resistance in Acinetobacter baumannii isolates collected from burn infection and Ventilator Associated Pneumonia (VAP).

Materials and methods: Ninety-eight A. baumannii isolates were collected from two different hospitals in Tehran, Iran. Tetracycline susceptibility testing was performed by disk diffusion and agar dilution methods according to the CLSI guidelines. The presence of adeSR, adeB, drug efflux system genes in resistant isolates was assessed by polymerase chain reaction (PCR). Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was used as a chemical inhibitor agent to assess the contribution of AdeABC efflux pump in tetracycline resistance isolates.

Results: Approximately 48% (47 out of 98) of isolates showed resistance to tetracycline which 14 (14.2%) isolates were corresponded to burn infection and the remaining 33 (33.8%) strains were isolated from VAP. All tetracycline resistant isolates have AdeABC in PCR assay. The reduction of tetracycline MICs by using 50 μg/ml CCCP were as follows: in 18 isolates 2-4 fold reduction in MICs, 26 isolates showed 8 fold reduction,1 isolate showed 16 fold, 1 isolate showed 32 fold and the remaining 1 isolate showed 128 fold reduction in MICs.

Conclusion: The results showed significant correlation between tetracycline resistance and AdeABC efflux pump genes in resistant A. baumannii isolates.

No MeSH data available.


Related in: MedlinePlus

PCR amplification of adeB, adeS and adeR genes in five selected isolates of A. baumannii. (a) Lane1: Positive control for adeB; lane 2-5: PCR product of adeB gene (541 bp); M: 1kb DNA size marker. (b) Lane1: Positive control for adeR; lane2-5: PCR product of adeR gene (447 bp); M: 1kb DNA size marker. (c) Lane1: positive control for adeS Lane 2-5: PCR product of adeS gene (544 bp); M: 1kb DNA size marker
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Figure 1: PCR amplification of adeB, adeS and adeR genes in five selected isolates of A. baumannii. (a) Lane1: Positive control for adeB; lane 2-5: PCR product of adeB gene (541 bp); M: 1kb DNA size marker. (b) Lane1: Positive control for adeR; lane2-5: PCR product of adeR gene (447 bp); M: 1kb DNA size marker. (c) Lane1: positive control for adeS Lane 2-5: PCR product of adeS gene (544 bp); M: 1kb DNA size marker

Mentions: PCR assay was performed with specific primers as described above. The expected amplification fragment with sizes of approximately 541 bp, 447 bp and 544 bp were detected on agaros gel corresponded to adeB, adeR and adeS genes, respectively in all tetracycline resistant isolates [Figure 1]. In the present study harboring the tetracycline resistance genes (AdeB, AdeR, AdeS) was confirmed in all resistant strains.


Detection of AdeABC efflux pump genes in tetracycline-resistant Acinetobacter baumannii isolates from burn and ventilator-associated pneumonia patients.

Beheshti M, Talebi M, Ardebili A, Bahador A, Lari AR - J Pharm Bioallied Sci (2014)

PCR amplification of adeB, adeS and adeR genes in five selected isolates of A. baumannii. (a) Lane1: Positive control for adeB; lane 2-5: PCR product of adeB gene (541 bp); M: 1kb DNA size marker. (b) Lane1: Positive control for adeR; lane2-5: PCR product of adeR gene (447 bp); M: 1kb DNA size marker. (c) Lane1: positive control for adeS Lane 2-5: PCR product of adeS gene (544 bp); M: 1kb DNA size marker
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231381&req=5

Figure 1: PCR amplification of adeB, adeS and adeR genes in five selected isolates of A. baumannii. (a) Lane1: Positive control for adeB; lane 2-5: PCR product of adeB gene (541 bp); M: 1kb DNA size marker. (b) Lane1: Positive control for adeR; lane2-5: PCR product of adeR gene (447 bp); M: 1kb DNA size marker. (c) Lane1: positive control for adeS Lane 2-5: PCR product of adeS gene (544 bp); M: 1kb DNA size marker
Mentions: PCR assay was performed with specific primers as described above. The expected amplification fragment with sizes of approximately 541 bp, 447 bp and 544 bp were detected on agaros gel corresponded to adeB, adeR and adeS genes, respectively in all tetracycline resistant isolates [Figure 1]. In the present study harboring the tetracycline resistance genes (AdeB, AdeR, AdeS) was confirmed in all resistant strains.

Bottom Line: All tetracycline resistant isolates have AdeABC in PCR assay.The reduction of tetracycline MICs by using 50 μg/ml CCCP were as follows: in 18 isolates 2-4 fold reduction in MICs, 26 isolates showed 8 fold reduction,1 isolate showed 16 fold, 1 isolate showed 32 fold and the remaining 1 isolate showed 128 fold reduction in MICs.The results showed significant correlation between tetracycline resistance and AdeABC efflux pump genes in resistant A. baumannii isolates.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Iran University of Medical Sciences, Tehran, Iran ; Department of Microbiology, Tehran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Purpose: Acinetobacter baumannii is the most prevalent nosocomial pathogen which have been emerged in the past three decades worldwide. The aim of this study was to assess the distribution of the AdeABC efflux pump genes, associated with tetracycline resistance in Acinetobacter baumannii isolates collected from burn infection and Ventilator Associated Pneumonia (VAP).

Materials and methods: Ninety-eight A. baumannii isolates were collected from two different hospitals in Tehran, Iran. Tetracycline susceptibility testing was performed by disk diffusion and agar dilution methods according to the CLSI guidelines. The presence of adeSR, adeB, drug efflux system genes in resistant isolates was assessed by polymerase chain reaction (PCR). Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was used as a chemical inhibitor agent to assess the contribution of AdeABC efflux pump in tetracycline resistance isolates.

Results: Approximately 48% (47 out of 98) of isolates showed resistance to tetracycline which 14 (14.2%) isolates were corresponded to burn infection and the remaining 33 (33.8%) strains were isolated from VAP. All tetracycline resistant isolates have AdeABC in PCR assay. The reduction of tetracycline MICs by using 50 μg/ml CCCP were as follows: in 18 isolates 2-4 fold reduction in MICs, 26 isolates showed 8 fold reduction,1 isolate showed 16 fold, 1 isolate showed 32 fold and the remaining 1 isolate showed 128 fold reduction in MICs.

Conclusion: The results showed significant correlation between tetracycline resistance and AdeABC efflux pump genes in resistant A. baumannii isolates.

No MeSH data available.


Related in: MedlinePlus