Development Of Bacterial Sensitivity Test Based On Quantitative Dna Assessment Using Uv Vis Spectroscopy

Abstract

Abstract-This research's objective is to develop methods of testing the sensitivity of bacteria to various antibiotics using quantitative assessment of DNA during log phase with UV-Vis Spectroscopy. Methods used, samples of pathogenic E. coli bacteria after cultured in broth solution containing Ciprofloxacin for 7 hours were taken for DNA extraction technique. The isolated DNA was then quantitatively calculated based on light absorbance on UV-Vis spectrophotometer, where the values of absorbance density was converted into the concentration of DNA/ml. As a control arm of this study, the equal sample of pathogenic E. coli bacteria were cultured in broth solution without antibiotic. Results: The DNA concentration from 15 samples of E. coli in broth culture solution containing antibiotic solution was 7.3 ± SD 2.28 ; whereas the DNA concentration in the control group was 10.3 ± SD 2.51 which was significant different (p. = 0. 000). This test was compared with conventional sensitivity test using disc diffusion method. It was demonstrated that the results were consistent with disc diffusion test that showing intermediate degree of sensitivity of the bacteria to the same antibiotic. These results indicate that administration of Ciprofloxacin in cultured pathogenic E. coli within the log phase of the bacteria inhibited the growth of DNA, which meant that the bacteria was sensitive to antibiotic. Conclusions: These results indicate that bacterial sensitivity test can be developed by DNA quantitative assessment during the growth of pathogenic bacteria using UV-Vis spectroscopy. This study will be followed by further sensitivity testing of various pathogenic bacteria to corresponding antibiotics for treatment of related disease.