Analysis of major band of Enterobacter sakazakii by ERIC-PCR and development of a species-specific PCR for detection of Ent. sakazakii in dry food samples

Journal of Microbiological Methods
Yingwang YeJumei Zhang

Abstract

ERIC (Enterobacterial Repetitive Intergenic Consensus)-PCR was employed to generate stable and reproductive ERIC-PCR fingerprints of Ent. sakazakii ATCC51329. Moreover, this study also cloned and sequenced a major band of Ent. sakazakii (ATCC51329) ERIC-PCR fingerprints. The major band was amplified with primer ERIC2 and sequences extending primer ERIC 2 showed poor similarity with ERIC elements. A comparison of the nucleotide acid with other sequences available in the GenBank revealed 90% of identity with Ent. sakazakii ATCC BAA-894, and 73%-74% of identity with oligopeptiase gene or protease gene of some species from the Enterobacteriaceae family. Two primers were synthesized to develop and optimize an Enterobacter sakazakii-specific PCR based on regions of major band unique to Ent. sakazakii. The expected fragment was amplified from all of Ent. sakazkaii but not from the negative controls. As few as 10(2) CFU/ml of Ent. sakazakii of PCR were directly detected in the infant formulas. This was the case even in the presence of other bacteria. A comparison of traditional methods and new developed PCR in commercial foods suggested that without using API20-E test, the DFI chromogenic medium and FDA method showed 46.15% and 50% fal...Continue Reading

References

Nov 25, 1990·Nucleic Acids Research·G J Sharples, R G Lloyd
Nov 15, 2002·International Journal of Food Microbiology·Marija Trkov, Gorazd Avgustin
May 7, 2003·Applied and Environmental Microbiology·Sandra L McLellanAlissa K Salmore
Sep 15, 2004·International Journal of Food Microbiology·Carol IversenStephen Forsythe
Nov 6, 2004·Journal of Clinical Microbiology·Carol IversenStephen Forsythe
Jul 26, 2005·International Journal of Food Microbiology·Joshua B GurtlerLarry R Beuchat
Oct 18, 2005·Molecular and Cellular Probes·Yin LiuXitai Huang
May 30, 2006·International Journal of Food Microbiology·Denise DrudySéamus Fanning

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Citations

May 12, 2009·International Journal of Food Microbiology·Yingwang YeJumei Zhang

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