Detection of Salmonella typhimurium in raw meats using in-house prepared monoclonal antibody coated magnetic beads and PCR assay of the fimA gene.
A method for detection of Salmonella Typhimurium in meat samples that uses in-house monoclonal antibody (MAb) coated magnetic beads for immunomagnetic separation (IMS) associated with PCR amplification of the gene fimA was developed. An internal amplification control (IAC) of the PCR reaction was constructed. The fimA PCR has shown 100% sensitivity and specificity when tested with various bacteria. The detection limit of the IMS-PCR method, using a post-enrichment in BHI broth for 6 h between IMS and PCR, was 1-10 CFU/mL. The method proved to be rapid (27 hrs), highly sensitive (1-10 CFU/25 g), and specific for detection of S. Typhimurium from experimentally contaminated pork and chicken meat samples.
Moreira
Angela N
AN
Centro de Biotecnologia, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Conceição
Fabricio R
FR
Conceição
Rita de Cássia S
Rde C
Ramos
Roberta J
RJ
Carvalhal
José B
JB
Dellagostin
Odir A
OA
Aleixo
José A G
JA
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D052203:Ferrosoferric Oxide / Q000737:chemistry; D036781:Fimbriae Proteins / Q000235:genetics; D005516:Food Microbiology; D018189:Immunomagnetic Separation / Q000379:methods*; D008460:Meat / Q000382:microbiology*; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods*; D012486:Salmonella typhimurium / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004269:DNA, Bacterial; C110675:SEF21 protein, Salmonella enteritidis; D036781:Fimbriae Proteins; D052203:Ferrosoferric Oxide