Detecção de genes de resistência à antimicrobianos em bactérias do grupo ESKAPE, isoladas de animais domésticos, por meio da técnica em amplificação isotérmica mediada por loop

Abstract

Antimicrobial resistance is an obstacle in the treatment of bacterial infections, requiring faster tests to define assertive and timely therapies. Aminoglycosides and β-lactams are important lines of treatment for bacterial infections related to the ESKAPE group bacteria, and the detection of resistance genes is significant for treatment efficiency. The objective of this study was to use the Loop-Mediated Isothermal Amplification (LAMP) technique to detect the antimicrobial resistance genes armA and blaKPC-2 in bacterial isolates from different lesion sites, collected from dogs, cats, cattle, horses, pigs and chickens, due to their potential as reservoirs of resistance genes. A total of 88 samples received at the Microbiology Laboratory of the Veterinary Hospital of UFMT, Cuiabá campus, were processed, obtaining 102 isolates, identified morphotinctorially and biochemically and submitted to antibiogram by disk diffusion. Of these isolates, 51 were gram-negative and 42 (82.45%) were multidrug-resistant. The detection of the armA gene was observed in 14 isolates out of a total of 30 (46%), resistant to aminoglycosides and 15 were negative in qPCR, out of a total of 18 sensitive (83.33%). Regarding blaKPC, 10 positive samples out of 15 resistant to carbapenems and 31 negative out of a total of 36 sensitive were detected, with both genes in three isolates and the highest number of positive bacteria related to Pseudomonas sp. (31.25%). The detection of the armA and blaKPC-2 genes in LAMP agreed with qPCR in 100% (3/3) for positive samples and 100% (5/5) for negative samples. Subsequently, clinical samples were tested, where urine and tissues showed the highest detection rate. It is concluded that this is an effective, less expensive and agile technique for diagnosing antimicrobial resistance genes in both bacterial isolates and clinical samples. Keywords: blaKPC, armA, genotypic method.