Open AccessInhibitory Effects of Collagen on the PCR for Detection of Clostridium perfringens
Author(s) -
Sangburm Kim,
Ronald G. Labbé,
Sangryeol Ryu
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.3.1213-1215.2000
Subject(s) - clostridium perfringens , dna extraction , polymerase chain reaction , dna , microbiology and biotechnology , real time polymerase chain reaction , chemistry , clostridium , extraction (chemistry) , biology , chromatography , bacteria , gene , biochemistry , genetics
It is essential to identify specific food components that inhibit PCR in order to increase the sensitivity of the PCR method for rapid detection of pathogens contaminating a food. We found that collagen, a major component of several foods, inhibited PCR. The inhibitory action of collagen on PCR could be partially reversed by adjusting the concentration of magnesium ion in the reaction mixture and by the use of various DNA extraction methods to remove the collagen from the DNA. Also, the source of thermostable DNA polymerase was affected by the presence of collagen. These results suggest the need to optimize the extraction and assay conditions for rapid detection of enterotoxigenicClostridium perfringens by PCR with respect to the kind of food being analyzed.