Rapid detection of Listeria monocytogenes by a PCR assay specific for an aminopeptidase.
Specific and rapid detection of Listeria monocytogenes is very important with regard to food safety since all other species of Listeria appear to be non-pathogenic to humans. Conventional microbiological detection methods are very time consuming. The polymerase chain reaction (PCR) is one of the most promising techniques for rapid detection of micro-organisms in food products. We have developed a PCR assay, specific for L. monocytogenes, based on the gene encoding an aminopeptidase, which previously has not been described for this species. The L. monocytogenes aminopeptidase shares strong sequence similarity with aminopeptidase C from Streptococcus thermophilous, Lactobacillus lactis, Lactobacillus helveticus, and with a cysteine proteinase from Saccharomyces cerevisiae. Polymerase chain reaction primers were synthesized based on the DNA sequence of the aminopeptidase gene. A 90 bp product was apparent with all L. monocytogenes strains tested but not with other species of Listeria or other bacterial genera. The PCR assay, which is performed directly from whole bacterial cells, does not involve DNA purification and can be conducted in 4 h. It provided positive identification of L. monocytogenes in mixed culture.
Winters
D K
DK
Department of Food Science and Center for Food Safety and Quality-IFSE, University of Arkansas, 272 Young Avenue, Fayetteville, AR 72704, USA. dwinters@comp.uark.edu
Maloney
T P
TP
Johnson
M G
MG
D000626:Aminopeptidases / Q000235:genetics*; D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D005516:Food Microbiology; D008087:Listeria / Q000201:enzymology / Q000235:genetics; D008089:Listeria monocytogenes / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D001426:Bacterial Proteins; D000626:Aminopeptidases