A Simple and Sensitive Polymerase Chain Reaction Method for the Detection of Food-Related Bacteria

Abstract

We devised a simple and efficient method of preparing chromosomal DNA from Gram-positive bacteria for use as a template in PCR, which consists of achromopeptidase treatment followed by Chelex 100 treatment. Using template DNA prepared by this method, it was possible to detect as few as one colony forming unit of some kinds of food-related bacteria using universal primers targeting the bacterial 16S ribosomal RNA gene or specific primers for bacterial genes under fixed thermocycling conditions.