OPTIMASI SUHU ANNEALING PROSES PCR AMPLIFIKASI GEN shv BAKTERI Escherichia coli PASIEN ULKUS DIABETIK

Abstract

Escherichia coli bacteria produce Extended Spectrum Beta Lactamase (ESBLs) to
hydrolyze beta lactam ring from beta-lactam antibiotics. One of the ESBLs coding genes
responsible for resistance is the shv or sulphydryl variable. Annealing temperature optimization
in the Polymerase Chain Reaction (PCR) process was carried out aimed at the success in
amplifying shv genes from E. coli bacteria. The method used in in vitro-based molecular testing
uses DNA Extraction Kit, PCR amplification and electrophoresis. The annealing temperature used
for pasting primer shv 5 'GGTTATGCGTTATATTCGCC 3' and reverse shv 5 primer
'TTAGCGTTGCCAGTGCTC 3' are 50 ° C, 54 ° C, 56 ° C, 57 ° C, 60 ° C and 62 ° C.
Visualization of the results of the amplification using a UV lamp at a wavelength of 254 nm did
not produce an amplicon with a size of 867 bp. The conclusion of this study shows that the
annealing temperature used is not yet optimum to identify shv genes from E. coli bacterial isolates
in diabetic ulcer patients who are resistant to antibiotics.