Evaluation of Primers Detection Capabilities of the pef Salmonella typhimurium Gene and the fimC Eschericia coli Gene Using Real-Time PCR to Develop Rapid Detection of Food Poisoning Bacteria

Abstract

The aim of the research is to obtain information about detection capabilities of the pef S. typhimurium and the fim C E. coli primer using Real time PCR method to develop kit detection of food poisoning bacteria. Evaluation of primer's capability was determined by accumulation of fluorescent signal in amplification curves that are connected by Cycle threshold (Ct) and intensity of amplicon signal. The results show the pef primer gene amplifies DNA target of the S. typhimurium fragments in Ct 17.85 and the fimC E. coli primer gene in Ct 12.25. Sensitivity evaluation of the pef S. typhimurium primer and the fimC E. coli primer showed a minimum detection level on 0,284 pg/μL and 7,12 pg/μL. Specificity evaluation of the pef gene of S. typhimurium primer on non-target E. coli bacteria showed Ct 18.05 almost similar to DNA target, but on primer fimC E. coli to non-target DNA bacteria S. typhimurium showed Ct 21.53. Therefore, it concluded the pef S. typhimurium primer gene and the fimC E. coli primer genes are suitable for rapid detection kit with good sensitivity. It needs further development in specificity of pef S. typhimurium primer, although the primer pef gene on non-target bacteria showed the different cycle threshold.