Detection of Salmonella Typhimurium in raw meats using in-house prepared monoclonal antibody coated magnetic beads and PCR assay of the fimA gene

被引:23
作者
Moreira, Angela N. [1 ,2 ]
Conceicao, Fabricio R. [1 ,3 ]
Conceicao, R. de C. S. [2 ,3 ]
Ramos, Roberta J. [1 ,2 ]
Carvalhal, Jose B. [2 ]
Dellagostin, Odir A. [1 ]
Aleixo, Jose A. G. [1 ,2 ]
机构
[1] Univ Fed Pelotas, Ctr Biotechnol, BR-96010900 Pelotas, RS, Brazil
[2] Univ Fed Pelotas, Fac Nutricao, Pelotas, RS, Brazil
[3] Univ Fed Pelotas, Fac Vet, Pelotas, RS, Brazil
关键词
Salmonella; foods; rapid methods; PCR; IMS; IAC;
D O I
10.1080/15321810701735096
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:58 / 69
页数:12
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