PENENTUAN SUBGENOTIPE GEN CTX-M PADA ESCHERICHIA COLI ISOLAT KLINIK RESISTEN SEFALOSPORIN GENERASI KETIGA

• Main Author: Natasha, Gabriela
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/48625
• Institution: Institut Teknologi Bandung

An infection is a disease caused by the entering of an infectious agent into the body. One method of treatment is the administration of antibiotics. Irrational use of antibiotics can cause an increase in bacterial resistance, especially resistance to third-generation cephalosporins in the Enterobacteriaceae family such as Escherichia coli. E. coli’s resistance to third-generation cephalosporins is mediated by the presence of the beta-lactamase enzyme, specifically the Extended- Spectrum Beta-Lactamase (ESBL) type, which is encoded by several types of genes, including the CTX- M gene. Genotypic differences cause phenotype changes, which can be observed in antibiotic resistance. This study aims to determine the CTX-M gene subgenotypes and the appropriate therapeutic recommendations for each subgenotype in order to ensure that the therapy given is optimal. This study uses a third-generation cephalosporin-resistant clinical isolates from one hospital in Bandung. Bacterial colonies from clinical isolates are rejuvenated in Luria Bertani (LB) media containing ceftriaxone. Bacteria are lysed using thermal cycler at 98 o C. The CTX-M gene is then detected in the lysed bacteria using PCR. PCR products are visualized by electrophoresis using 1% agarose gel. As many as 83.33% of clinical isolates (n = 30) positively contain the CTX-M gene. Some positive clinical isolates are chosen based on the intensity of the electropherogram for the DNA sequencing process. Sequencing results are then analyzed using nucleotide BLAST analysis. The result showed that 90% of clinical isolates (n = 10) contain the CTX-M-15 subgenotype (Accesion number: GQ274928) with a homology percentage of 99-100%, whilst 10% of clinical isolates (n = 10) contain the CTX-M-55 subgenotype (Accesion number: LC512850) with a homology percentage of 100%. The treatment recommendations for the CTX-M-15 subgenotype are carbapenem or piperacillintazobactam, whereas the treatment recommendation for the CTX-M-55 subgenotype is carbapenem.