STUDY OF MOLECULAR IDENTIFICATION USING MULTIPLEX PCR IN Salmonella sp. & Staphylococcus aureus DIAGNOSTICS CAUSING FOOD POISONING DISEASES
Abstract
The identification of infectious and opportunistic microorganism has recently been based on traditional culturing with low and cost effective, time controlling and hard manipulation for users and agencies. The new era of gene techniques that applied on observing, patrolling bacterial contamination to take minor times for prevention and medical cures have been considered as essential needs. The discovery of multiplex PCR could be applied in medical diagnostic because of quick returns, high sensitivity and parallel amplification of DNA bacteria for PCR results. This study indicates that the successful process of quick PCR amplification using multiplex PCR have been introduced for Salmonella sp. and Staphycoccus aureus. The optimal reaction of PCR have been chosen at the total volume of 25 µL with 2.5 µL 10X PCR buffer, 1.5 µL MgCl2 25 mM, 3 µL dNTPs 2.5 mM; 1 µL of invAF/R, 2.0 µL each primer of nucF/R respectively, 10 µL DNA templates và 1.8 µL dilution H2O. The highest sensitivity of multiplex PCR also showed and complied at 300 CFU/mL for Salmonella sp. and 70 CFU/mL for Staphylococcus aureus test. The essential time for poisonous sample culturing (1 CFU/25g sample) required at least 14 hours of growing culture in medium. The specific evaluation of this study for the other bacteria has been considered in larger scale from different types of foods and sources when applying multiplex PCR.
Keywords: Multiplex PCR, food poisoning, Salmonella sp., Staphylococcus aureus, invA, nuclease.