Identificación de bacterias multirresistentes a antibióticos y detección de genes de resistencia en pacientes pediátricos con gastroenteritis

Abstract

Antibiotics serve for the treatment and control of infectious diseases. Actually, the continuous development, the propagation of bacterial resistances and multiresistances pose problems about their future, representing a threat in the efficacy of treatments, as a consequence increasing morbimortality due to the capacity of dissemination in pathologies such as acute gastroenteritis (AGE), which has not been widely studied in our country. There is research on the presence of bacteria resistant and multiresistant to antibiotics in adults and children worldwide, but despite this, there is still no information and data on multiresistant bacteria in children with AGE in Ecuador. Objective: To determine the presence of multiresistant bacteria to antibiotics in pediatric patients with acute gastroenteritis, by means of microbiological and molecular techniques. Materials and methods: Bacterial isolates in stool of 206 pediatric patients with diagnosis of AGE will be analyzed, as inclusion criteria only those patients who have not received any antibiotic-based treatment. For the microbiological part, a protocol will be used as recommended by the Clinical Laboratory Standards Institute (CLSI) guidelines, in the phenotypic isolation of bacteria, agars such as MacConkey, SS Agar (Salmonella, Shigella) and Preston Agar will be used for bacterial identification, tests such as catalase and coagulase, biochemical tests such as TSI or Kligler, lysine, citrate, urea, SIM will be used. In antibiotic susceptibility will be determined by the disk diffusion method using the kirby-baur technique on Mueller-Hinton agar plates, the antibiotic disks to be used will be: Ceftriaxone (CRO)30 ug, Cefuroxime (CXM) 30 ug, Amikacin (AMK) 30 ug, Ampicillin+Sulbactam (SAM) 20 ug, Amoxicillin+ Clavulanic Acid (AMC) 20/10 ug, Gentamicin (GEN) 10 ug, Fosfomycin (FOS) 20 ug and Sulfatrimetropin (STX) 20 ug. For the molecular part, six sets of specific primers will be used to obtain PCR amplicons of resistance-associated genes (blaTEM-1, blaCTX-M, blaAmpC, FosA1, Sul-1 and aac (6') /aph (2')). The amplified products will be analyzed by agarose gel electrophoresis. Expected results: To find multiresistant bacteria in samples from pediatric patients with GEA such as E. coli, Salmonella, Shiguella, Klebsiella pneumoniae and Proteus. In addition to the molecular determination of resistance genes associated with microbiological phenotypic characteristics. The results obtained from this research will set a precedent in Ecuador by providing epidemiological data on the presence of multiresistant bacteria in pediatric populations. Further research will be needed to determine whether the resistance genes found are heritable or not.