Detection of ESBL and plasmid-mediated AmpC beta lactamases among the Gram-negative bacterial isolates in diabetic foot ulcer infections
ESBL and plasmid mediated AmpC in diabetic foot ulcer
Keywords:
AmpC, diabetic foot, ESBL, MICAbstract
Background and Aim: Foot ulcers are a significant complication of diabetes mellitus and are usually poly-microbial. Aerobic Gram-negative bacilli are isolated in higher frequency with increasing grade of ulcers, and development of drug resistance is a cause of concern. The aim of our study is to identify the extended spectrum β-lactamases (ESBL) and AmpC β-lactamases among the Gram-negative aerobic bacterial isolates in the different grades of diabetic foot ulcers. Materials and Methods: Pus samples from 104 male and female diabetic patients presenting with grade I to grade V foot ulcers were cultured according to the standard microbiological procedures and antimicrobial sensitivity performed by the Kirby-Bauer disc diffusion method as per CLSI guidelines. MIC 50 of all isolates for Ceftazidime was detected by the agar dilution method. The Gram-negative bacterial isolates were further tested for ESBL and AmpC β-lactamases by the modified double disc synergy test and combined disc method. AmpC disc test was used for the confirmation of AmpC production. Results: Forty-seven isolates exhibited resistance to Ceftazidime phenotypically and 54 isolates had MIC greater than 2 μg/mL. ESBL production was observed in 55.55% of the isolates. AmpC production was seen in 19 isolates (35.18%). No mechanism of cephalosporin resistance was explainable for 5 isolates. Conclusion: The combined disc method was more sensitive in the detection of ESBLs, while the AmpC disc test was an easy way to detect AmpC beta lactamases and can be used for routine screening.
References
Halpati A, Desai KJ, Jadeja R, Parmar M. A study of aerobic and anaerobic bacteria in diabetic foot ulcer and in vitro sensitivity of antimicrobial agent. Int J Med Sci Public Health 2014;3.
Raymundo MF, Mendoza MT. The microbiologic features and clinical outcome of diabetic foot infections among patients admitted at UP-PGH. Phil J Microbiol Infect Dis 2002;31:51-63.
Citron DM, Goldstein EJ, Merriam CV, Lipsky BA, Abramson MA. Bacteriology of moderate-to-severe diabetic foot infections and in vitro activity of antimicrobial agents. J Clin Microbiol 2007;45:2819-28.
Rouhipour N, Hayatshahi A, Nikoo MK, Yazdi NM, Heshmat R, Qorbani M, et al. Clinical microbiology study of diabetic foot ulcer in Iran; pathogens and antibacterial susceptibility. Afr J Microbiol Res 2012;6:5601-8.
Tiwari S, Pratyush DD, Dwivedi A, Gupta SK, Rai M, Singh SK. Microbiological and clinical characteristics of diabetic foot infections in northern India. J Infect Dev Ctries 2012;6:329-32.
Gadepalli R, Dhawan B, Sreenivas V, Kapil A, Ammini AC, Chaudhary R. A clinico-microbiological study of diabetic foot ulcers in an Indian tertiary care hospital. Diabetes Care 2006;29:1727-32.
CLSI, Performance standards for antimicrobial susceptibility testing; Twenty second informational supplement. CLSI document M100-S22. Wayne: Clinical and Laboratory Standards Institute; 2012.
Winn W, Allen S, Janda W, Koneman E, Procop G, Schreckenberger P, et al. Antimicrobial susceptibility testing. In: Koneman′s colour atlas and textbook of diagnostic microbiology. 6 th ed. Philadelphia: Lippincott William and Wilkins; 2006. p. 961-2.
Jog AS, Shadija PG, Ghosh SJ. Detection of multidrug resistant gram negative bacilli in type II diabetic foot infection. Int J Med Health Sci 2013;2:186-94.
Numanovic F, Hukic M, Deliegovic Z, Tihic N, Pasic S, Gegic M. Comparison of double disc synergy test, Vitek 2 and Check MDR CT102 for detection of ESBL producing isolates. Acta Med Acad 2013;42:15-24.
Thomson KS. Controversies about extended spectrum and AmpC Beta-Lactamases. Emerg Infect Dis 2001;7:333-6.
Dhara M, Disha P, Sachin P, Manisha J, Seema B, Vegad MM. Comparison of various methods for the detection of Extended spectrum beta lactamases in Klebsiella pneumoniae isolated from neonatal intensive care units, Ahmedabad. Nordic J Migrat Res 2012;2:348-53.
Goyal A, Prasad A, Ghosal U, Prasad KN. Comparison of disc diffusion, disc potentiation and double disc synergy methods for the detection of Extended spectrum beta lactamases in Enterobacteriaceae. Indian J Med Res 2008;128:209-11.
Varaiya AY, Dogra JD, Kulkarni MH, Bhalekar PN. Extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae in diabetic foot infections. Indian J Pathol Microbiol 2008;51:370-2.
Black JA, Moland ES, Thomson KS. AmpC disc test for detection of plasmid mediated AmpC beta lactamases in Enterobacteriaceae lacking chromosomal AmpC beta lactamases. J Clin Microbiol 2005;43:3110-3.
Yong D, Park R, Yum JH, Lee K, Choi EC, Chong Y. Further modification of the Hodge test to screen AmpC beta lactamases (CMY-1) producing strains of Escherichia coli and Klebsiella pneumoniae. J Microbiol Methods 2002;51:407-10.
Coudron PE. Inhibitor based methods for detection of plasmid mediated AmpC beta lactamases in Klebsiella spp, Escherichia coli and Proteus mirabilis. J Clin Microbiol 2005;43:4163-7.
Singhal S, Mathur T, Khan S, Upadhyay DJ, Chugh S, Gaind R, et al. Evaluation of methods for AmpC beta lactamases in gram negative clinical isolates from tertiary care hospitals. Indian J Med Microbiol 2005;23:120-4.
Sinha P, Sharma R, Rishi S, Sharma R, Sood S, Pathak D. Prevalence of extended spectrum beta lactamase producers among Escherichia coli isolates in a tertiary care hospital in Jaipur. Indian J Pathol Microbiol 2008;51:367-9.
Rand KH, Turner B, Seifert H, Hansen C, Johnson JA, Zimmer A. Clinical laboratory detection of AmpC β-lactamase: Does it affect patient outcome? Am J Clin Pathol 2011;135:572-6.
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