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1.
Biosens Bioelectron ; 25(5): 967-71, 2010 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-19913402

RESUMO

An SPR-based sandwich immunoassay for serotyping of Salmonella is demonstrated. The Salmonella are captured on an SPR chip using polyclonal capture antibody. SPR sensorgrams are obtained for the immunoreactions of the somatic (O) and flagellar (H) surface antigens, of the captured bacteria, to their respective antibodies. The sensorgram data are compiled to determine the antigenic formula in accordance with the Kauffmann-White scheme. Salmonella Enteritidis was completely serotyped using this SPR-based method. In addition, Salmonella belonging to serogroups B, C and D were successfully assigned to their respective serogroups. Before serotyping the bacteria are grown to a concentration of 1x10(10) mL(-1). This SPR-based serotyping provides quantitative data, and thus, eliminates the possibility of false detections as encountered in the conventional slide agglutination test (SAT). This method was also proved to work with rough strains.


Assuntos
Técnicas Biossensoriais/instrumentação , Contagem de Colônia Microbiana/instrumentação , Imunoensaio/instrumentação , Salmonella/isolamento & purificação , Sorotipagem/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Reprodutibilidade dos Testes , Salmonella/imunologia , Sensibilidade e Especificidade
2.
J Microbiol Methods ; 75(3): 545-50, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18804126

RESUMO

We present a rapid surface plasmon resonance-based serological assay for the detection of Salmonella Typhimurium infection in pigs using the Plasmonic((R)) SPR device. Lipopolysaccharide (LPS, 10 microg mL(-1)) from Salmonella Typhimurium was immobilised by self-assembly on a hydrophobic SPR chip. Using this LPS-coated chip, it was possible to bind and detect the anti-Salmonella Typhimurium antibodies in serum of pigs infected with the bacteria. The developed SPR assay is able to differentiate between sera obtained from pigs having low, medium, and high levels of Salmonella infection. A commercial ELISA kit was used to classify the sera for levels of Salmonella infection on the basis of optical density (OD%). A strong positive correlation was observed between the SPR-based assay and the ELISA (n=38, r=0.90, p<0.01). The sensitivity and specificity of the assay are 0.93 and 0.87, respectively. The SPR-based assay is label-free and does not require any sample preparation or dilution steps. The total analysis time is 45 min for each serum sample. The assay was found to be specific for Salmonella Typhimurium and shows no cross-reactivity to Salmonella Choleraesuis or Escherichia coli antibodies. As no sample preparation is required the developed assay has the potential to be used as a reliable tool for Salmonella monitoring programmes in pork production.


Assuntos
Salmonelose Animal/imunologia , Salmonella/isolamento & purificação , Testes Sorológicos/métodos , Ressonância de Plasmônio de Superfície/métodos , Doenças dos Suínos/imunologia , Animais , Anticorpos Antibacterianos/sangue , Salmonella/imunologia , Salmonelose Animal/microbiologia , Suínos , Doenças dos Suínos/microbiologia
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