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Journal of Clinical Microbiology, June 2006, p. 2199-2206, Vol. 44, No. 6
0095-1137/06/$08.00+0     doi:10.1128/JCM.00344-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Rapid Discrimination of Listeria monocytogenes Strains by Microtemperature Gradient Gel Electrophoresis

Saitama Industrial Technology Center North Institute, 2-133, Suehiro, Kumagaya, Saitama 360-0031, Japan

Received 16 February 2006/ Accepted 27 March 2006

Microtemperature gradient gel electrophoresis (µ-TGGE) was examined for use for the rapid subtyping of Listeria monocytogenes strains. Comparison of genomes between L. monocytogenes strains F2365 and H7858 identified a sequence encoding a portion of the PRT/PTS system IIA 2 protein domain as appropriate for µ-TGGE analysis. Thirty-one strains belonging to 10 different serovar types were tested by PCR, and sequence analysis of the amplified products revealed that the strains comprise 11 groups. All 55 possible pairs within the 11 groups were examined by µ-TGGE analysis. Of these, 47 pairs could be successfully discriminated, with a total electrophoresis time of only 7 min. Moreover, Cy3/Cy5 labeling allowed rapid identification of the sequence type in unknown strains of L. monocytogenes isolated from meat. These findings collectively indicate that µ-TGGE can be used for the rapid analysis of L. monocytogenes strains, facilitating determination of routes of contamination when these bacteria are found in food products.