Detection of enteroaggregative Escherichia coli by loop-mediated isothermal amplification.
A novel gene amplification method, loop-mediated isothermal amplification (LAMP), has been recently developed as a rapid, specific diagnostic method for various infectious diseases. We have investigated whether LAMP can be used to detect small numbers of enteroaggregative Escherichia coli (EAEC) cells contaminated in food samples. Primers for LAMP reaction were designed with EAEC aggR gene sequences (available in GenBank). LAMP specificity with these primers was the same as that of PCR in a study of 37 EAEC and 42 non-EAEC bacterial strains. The sensitivity of the LAMP method was better than that of PCR in a study of serially diluted EAEC cells. The LAMP method was significantly more effective than was PCR in detecting EAEC-contaminated food samples (Fisher's exact test, P < 0.05). Therefore, the LAMP method described here should be useful for detecting small numbers of EAEC cells in food samples.
Division of Bacteriology, Chiba Prefectural Institute of Public Health, 666-2, Chuo, Chiba City, Chiba 260-8715, Japan. e.ykym@ma.pref.chiba.lg.jp
Eiji
E
Yokoyama
Masako
M
Uchimura
Kenitiro
K
Ito
D001483:Base Sequence; D003257:Consumer Product Safety; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial