Phenotypic Detection of Metallo-Β Lactamase Producing Carbapenem-Resistant Pseudomonas Aeruginosa

Document Type : Original Article

Authors

1 Department of Microbiology, Faculty of Medicine, Umm Al-Qura University, Makkah, Saudi Arabia.

2 Department of Agricultural Sciences, Food, Natural Resources and Engineering, Università degli Studi di Foggia, Italy

Abstract

The opportunistic bacteria Pseudomonas aeruginosa is commonly linked to skin infections. When dealing with P. aeruginosa, carbapenem is your best bet. It is of international concern because beta-lactamase synthesis might lead to carbapenem resistance. Among all the beta-lactamases, Metallo-Beta lactamases are the most versatile. The purpose of this study was to isolate metallo—lactamase-producing P. aeruginosa from wound infections. One hundred and twenty samples from patients with burn wound infections were collected and tested using conventional microbiological methods for the presence of P. aeruginosa. Using species-specific primers against the oprL and Oprl genes of P. aeruginosa, a polymerase chain reaction was used to conduct a molecular characterisation of P. aeruginosa. Antimicrobial susceptibility testing was conducted using the Kirby Bauer disc diffusion technique. Certain primers were used in a polymerase chain reaction to detect P. aeruginosa carrying the carbapenemase gene. Using the modified carbapenem inactivation technique and the EDTA carbapenem inactivation approach, we were able to phenotypically identify metallo-beta-lactamase-expressing genes. Carbapenemase-encoding genes' sensitivities and specificities were determined. Forty-six out of a possible one hundred P. aeruginosa (38%) were successfully recovered, with their identity validated by a polymerase chain reaction (PCR) experiment. The highest level of resistance was discovered against Cefepime (87%) and the lowest level of resistance was recorded against colistin (33%). A total of 25 (54%) were multidrug-resistant isolates. Out of 46, 35(76%) were confirmed for carbapenemase production by performing PCR. The prevalence of carbapenemase encoding genes was as follows: blaSPM (14%), blaVIM (25.7%), blaNDM (40%), blaKPC (2.85%) and blaIMP (17%).  The modified carbapenem inactivation method showed 91.42% positive results and eCIM showed 90.62% positive results. Phenotypic detection showed more sensitivity and less specificity. Results concluded that mCIM and eCIM were considered a less expensive, more sensitive and suitable method to distinguish class A and class B of carbapenemase-producing P. aeruginosa.

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