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Characterization of multidrug resistant ESBL-producing Escherichia coli isolates from hospitals in Malaysia.

Lim KT, Yasin R, Yeo CC, Puthucheary S, Thong KL - J. Biomed. Biotechnol. (2009)

Bottom Line: Amplification and sequence analysis of the 5'CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons.Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible.These isolates were very diverse and heterogeneous.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Institute of Biological Science, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
The emergence of Escherichia coli that produce extended spectrum beta-lactamases (ESBLs) and are multidrug resistant (MDR) poses antibiotic management problems. Forty-seven E. coli isolates from various public hospitals in Malaysia were studied. All isolates were sensitive to imipenem whereas 36 were MDR (resistant to 2 or more classes of antibiotics). PCR detection using gene-specific primers showed that 87.5% of the ESBL-producing E. coli harbored the blaTEM gene. Other ESBL-encoding genes detected were blaOXA, blaSHV, and blaCTX-M. Integron-encoded integrases were detected in 55.3% of isolates, with class 1 integron-encoded intI1 integrase being the majority. Amplification and sequence analysis of the 5'CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons. Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible. All 47 isolates were subtyped by PFGE and PCR-based fingerprinting using random amplified polymorphic DNA (RAPD), repetitive extragenic palindromes (REPs), and enterobacterial repetitive intergenic consensus (ERIC). These isolates were very diverse and heterogeneous. PFGE, ERIC, and REP-PCR methods were more discriminative than RAPD in subtyping the E. coli isolates.

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(a) Representative ERIC-PCR profiles for E. coli  isolates. Lanes 1, 10, 20: 100 bp DNA ladder, lanes 2–4: EC1-EC3; lanes 5–9: EC17, EC19, EC18, EC30, EC34, lanes 11–18: EC32, EC14, EC41, EC39, EC42, EC22, EC45, EC46; lane 19: negative control. (b) Representative RAPD profiles for E. coli   isolates. Lanes 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC40, EC26, EC43, EC46, EC47;  lane 19: negative control. (c) Representative REP-PCR profiles for E. coli  isolates. Lane 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC26, EC40, EC43, EC46, EC47; lane 19: negative control. (d) Representative PFGE profiles for E. coli  isolates. Lanes 1, 7, 14: Salmonella  Braenderup H9812 Standard DNA marker; lanes 2–6: EC4, EC9, EC38, EC36, EC18; lanes 8–13: EC28, EC31, EC40, EC22, EC45, EC46.
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fig1: (a) Representative ERIC-PCR profiles for E. coli isolates. Lanes 1, 10, 20: 100 bp DNA ladder, lanes 2–4: EC1-EC3; lanes 5–9: EC17, EC19, EC18, EC30, EC34, lanes 11–18: EC32, EC14, EC41, EC39, EC42, EC22, EC45, EC46; lane 19: negative control. (b) Representative RAPD profiles for E. coli isolates. Lanes 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC40, EC26, EC43, EC46, EC47; lane 19: negative control. (c) Representative REP-PCR profiles for E. coli isolates. Lane 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC26, EC40, EC43, EC46, EC47; lane 19: negative control. (d) Representative PFGE profiles for E. coli isolates. Lanes 1, 7, 14: Salmonella Braenderup H9812 Standard DNA marker; lanes 2–6: EC4, EC9, EC38, EC36, EC18; lanes 8–13: EC28, EC31, EC40, EC22, EC45, EC46.

Mentions: Three PCR-based DNA fingerprinting methods were used to subtype the 47 E. coli isolates. ERIC-PCR analysis differentiated the 47 isolates into 45 unique profiles (F = 0.54–1.0) whereas RAPD using the OPAB04 and OPB17 primers generated 44 and 43 profiles, respectively, (F = 0.41–1.0 for the OPAB04 primer and F = 0.36–1.0 for the OPB17 primer, see Figures 1(a) and 1(b)). REP-PCR differentiated the 47 isolates into 45 distinct profiles (F = 0.53–1.0, see Figure 1(c)). All three PCR-based methods were reproducible as identical profiles were obtained in separate experiments using the same set of isolates.


Characterization of multidrug resistant ESBL-producing Escherichia coli isolates from hospitals in Malaysia.

Lim KT, Yasin R, Yeo CC, Puthucheary S, Thong KL - J. Biomed. Biotechnol. (2009)

(a) Representative ERIC-PCR profiles for E. coli  isolates. Lanes 1, 10, 20: 100 bp DNA ladder, lanes 2–4: EC1-EC3; lanes 5–9: EC17, EC19, EC18, EC30, EC34, lanes 11–18: EC32, EC14, EC41, EC39, EC42, EC22, EC45, EC46; lane 19: negative control. (b) Representative RAPD profiles for E. coli   isolates. Lanes 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC40, EC26, EC43, EC46, EC47;  lane 19: negative control. (c) Representative REP-PCR profiles for E. coli  isolates. Lane 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC26, EC40, EC43, EC46, EC47; lane 19: negative control. (d) Representative PFGE profiles for E. coli  isolates. Lanes 1, 7, 14: Salmonella  Braenderup H9812 Standard DNA marker; lanes 2–6: EC4, EC9, EC38, EC36, EC18; lanes 8–13: EC28, EC31, EC40, EC22, EC45, EC46.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig1: (a) Representative ERIC-PCR profiles for E. coli isolates. Lanes 1, 10, 20: 100 bp DNA ladder, lanes 2–4: EC1-EC3; lanes 5–9: EC17, EC19, EC18, EC30, EC34, lanes 11–18: EC32, EC14, EC41, EC39, EC42, EC22, EC45, EC46; lane 19: negative control. (b) Representative RAPD profiles for E. coli isolates. Lanes 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC40, EC26, EC43, EC46, EC47; lane 19: negative control. (c) Representative REP-PCR profiles for E. coli isolates. Lane 1, 21: 1 kb DNA ladder; lane 2, 20: 100 bp DNA ladder; lanes 3–18: EC5, EC8, EC13, EC15, EC7, EC44, EC12, EC24, EC25, EC29, EC35, EC26, EC40, EC43, EC46, EC47; lane 19: negative control. (d) Representative PFGE profiles for E. coli isolates. Lanes 1, 7, 14: Salmonella Braenderup H9812 Standard DNA marker; lanes 2–6: EC4, EC9, EC38, EC36, EC18; lanes 8–13: EC28, EC31, EC40, EC22, EC45, EC46.
Mentions: Three PCR-based DNA fingerprinting methods were used to subtype the 47 E. coli isolates. ERIC-PCR analysis differentiated the 47 isolates into 45 unique profiles (F = 0.54–1.0) whereas RAPD using the OPAB04 and OPB17 primers generated 44 and 43 profiles, respectively, (F = 0.41–1.0 for the OPAB04 primer and F = 0.36–1.0 for the OPB17 primer, see Figures 1(a) and 1(b)). REP-PCR differentiated the 47 isolates into 45 distinct profiles (F = 0.53–1.0, see Figure 1(c)). All three PCR-based methods were reproducible as identical profiles were obtained in separate experiments using the same set of isolates.

Bottom Line: Amplification and sequence analysis of the 5'CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons.Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible.These isolates were very diverse and heterogeneous.

View Article: PubMed Central - PubMed

Affiliation: Microbiology Division, Institute of Biological Science, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
The emergence of Escherichia coli that produce extended spectrum beta-lactamases (ESBLs) and are multidrug resistant (MDR) poses antibiotic management problems. Forty-seven E. coli isolates from various public hospitals in Malaysia were studied. All isolates were sensitive to imipenem whereas 36 were MDR (resistant to 2 or more classes of antibiotics). PCR detection using gene-specific primers showed that 87.5% of the ESBL-producing E. coli harbored the blaTEM gene. Other ESBL-encoding genes detected were blaOXA, blaSHV, and blaCTX-M. Integron-encoded integrases were detected in 55.3% of isolates, with class 1 integron-encoded intI1 integrase being the majority. Amplification and sequence analysis of the 5'CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons. Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible. All 47 isolates were subtyped by PFGE and PCR-based fingerprinting using random amplified polymorphic DNA (RAPD), repetitive extragenic palindromes (REPs), and enterobacterial repetitive intergenic consensus (ERIC). These isolates were very diverse and heterogeneous. PFGE, ERIC, and REP-PCR methods were more discriminative than RAPD in subtyping the E. coli isolates.

Show MeSH
Related in: MedlinePlus