Rapid detection of salmonellae in poultry with the magnetic immuno-polymerase chain reaction assay.
Rapid detection of salmonellae in chicken meat was accomplished by using the magnetic immuno-polymerase chain reaction assay (MIPA). A direct polymerase chain reaction assay performed with chicken meat spiked with Salmonella typhimurium resulted in poor sensitivity (approximately 10(7) CFU/g of meat). The use of immunoseparation with a Salmonella serogroup B-specific monoclonal antibody improved the sensitivity, but enrichment was required for the detection of low levels of contamination. Enrichment for 6 h in either buffered peptone water, lactose broth containing tergitol-7, or selenite-cystine broth resulted in the detection of an initial inoculum of 100 CFU per g of meat. Enrichment of the salmonellae present on 25 g of spiked chicken meat for 24 h in either buffered peptone water or selenite-cystine broth before detection by the MIPA yielded a detection limit of approximately 0.1 CFU/g of meat. A detection limit of approximately 1 CFU/g of meat was obtained when the spiked meat was stored at -20 degrees C before enrichment for 24 h and analysis with the MIPA. Although the MIPA was developed for S. typhimurium, a MIPA in which a panel of six monoclonal antibodies specific for Salmonella serogroups A through E was used detected the presence of 0.1 CFU of Salmonella enteritidis per g of chicken meat. These data indicate that the method is applicable to other commonly isolated serotypes.
Eijkman-Winkler Laboratory for Medical Microbiology, University Hospital Utrecht, The Netherlands.
A C
AC
Fluit
M N
MN
Widjojoatmodjo
A T
AT
Box
R
R
Torensma
J
J
Verhoef
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002645:Chickens / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D005069:Evaluation Studies as Topic; D005516:Food Microbiology; D008280:Magnetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012475:Salmonella / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D004269:DNA, Bacterial