Detection of Antibiotic-Resistant Gene from Isolates of Diabetic Ulcer Patients

Abstract

Antibiotic resistance can be caused by the presence of several genes encoding antibiotic resistance found in certain bacteria. S. aureus bacteria contained blaZ (97.4%), mecA (42.3%) and tetM genes (20.2%). Bacillus sp. contained blm (87.2%), tetL (44%) and tetB genes (2.9%). E. coli bacteria contained tem (80.9%)dan shv genes (14.28%). Clostridium spp. Bacteria contained cat gene (36%), as well as S. pyogens contained ermT (36.5%). Specific primers made in silico based on bioinformatics were then analyzed by PCR (Polymerase Chain Reaction) and electrophoresis. The purpose of this study was to detect the presence of blaZ, mecA, tetM, blM, tetL, tetB, tem, shv, cat, and ermT genes. The research began by searching for the nucleotide sequences for the ermT, cat, tetB, and blaZ genes through the Primer 3Plus program and analyzed using OligoAnalyzer 3.1. In vitro detection was carried out by extracting DNA and then amplification using PCR, and analyzed by agarose gel electrophoresis. The results of the analysis of the primer criteria for the ermT gene and the blaZ gene at the optimum Tm 55 C, cat gene and the tetB gene at the optimum Tm 60 C have met all the primer criteria. The results obtained are mecA gene (533 bp), tetM gene (366 bp), blm gene (1107 bp), tetL gene (267 bp), tetB gene (186 bp), tem gene (1073 bp), cat gene (164 bp), the ermT gene (202 bp) has been detected successfully, while blaZ and shv have not been detected successfully.