Development and Application of an Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Prevalent <i>Salmonella</i> Serovars in Swine in Southern Brazil

Kich, Jalusa Deon; Schwarz, Patrícia; Silva, Luis Eduardo; Coldebella, Arlei; Piffer, Itamar Antônio; Vizzoto, R.; de, Itapema Cardoso Marisa Ribeiro

doi:10.1177/104063870701900508

Cited by 20 publications

(21 citation statements)

References 24 publications

(67 reference statements)

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“…Alternatively, ELISA for a specific bacteria antigen has been reported in the literature [12,13,14]. However, the drawback for this approach is that it will only specifically detect one type of bacteria and ELISA assay requires dedicated equipment such as ELISA plate-reader, plate-washer and skilled technical support.…”

Section: Discussionmentioning

confidence: 99%

Anti-LPS Test Strip for the Detection of Food Contaminated with Salmonella and E. coli

Wang¹,

Molina²,

Prima³

et al. 2011

JMBT

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Enterobacteria such as E. coli and Salmonella release an endotoxin, or lipopolysaccharide (LPS) that contributes to food poisoning symptoms. We hypothesize that LPS could be detected by a simple strip test, thus detecting the presence of enterobacteria in food. Here we described the development of a novel Anti-LPS detection strip test. Flexible polystyrene test strips with an absorbent poly (vinylidene fluoride) membrane adhered onto one end were exposed to various dilutions of food or bacterial sources. This was followed by the sequential incubation with a primary anti-LPS antibody, biotinylated secondary antibody, and streptavidin-linked enzyme alkaline phosphatase. The test strip was then developed with substrate 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium. Thus, the presence of purple precipitate product would indicate the presence of LPS and enterobacteria. The intensity or darkness of the color was quantified densitometrically and compared to LPS standard curve. The test strip assay was shown to readily detect as low as 25 ng/mL of purified LPS. In addition, theanti-LPS test strip assay can sensitively detect and quantify LPS released by live E. coli into culture media. Finally, three food groups (strawberry slices, spinach leaves and ground beef) were inoculated with E. coli for two time points at room temperature and then rinse water from each food preparation was subjected to the anti-LPS test strip assay. For all three food groups, the test strip assay can readily detect and quantify both 8 h and 24 h bacteria contamination over their respective uncontaminated controls. In conclusion, a simple prototype anti-LPS strip test was developed to readily detect enterobacteria contamination of common food.

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Section: Discussionmentioning

confidence: 99%

Anti-LPS Test Strip for the Detection of Food Contaminated with Salmonella and E. coli

Wang¹,

Molina²,

Prima³

et al. 2011

JMBT

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“…O aumento do número de animais infectados tem sido detectado nas fases de crescimento e terminação (KICH et al, 2007), justificando a aplicação de medidas de controle nessas fases. Entre as intervenções possíveis, o fornecimento, via ração, de aditivos que permitam a diminuição do número de suínos infectados ou que excretem Salmonella nas fezes, pode contribuir para alcançar um menor número de animais portadores ao abate.…”

Section: Control Of Salmonella Transmission Has Been a Challenge For unclassified

Efeito de probiótico na infecção e excreção fecal de Salmonella em suínos

et al. 2012

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A transmissão de Salmonella na cadeia produtiva de suínos é um problema de difícil controle. O objetivo do estudo foi avaliar o efeito da administração oral de probiótico sobre a ocorrência de infecção e excreção fecal de Salmonella em suínos em fase de crescimento. Os tratamentos consistiram de ração basal sem aditivos (controle) ou adicionada de probiótico (10(7)ufc g-1 de células viáveis dos gêneros Bifidobacterium, Enterococcus, Lactobacillus e Saccharomyces). Foram alocados seis leitões de 50 dias em cada tratamento, com duas repetições por tratamento. Todos os animais foram inoculados com Salmonella Typhimurium (10(6)ufc mL-1) após 14 dias do alojamento. Semanalmente, foram coletadas amostras de sangue e fezes e no dia 35 pós-inoculação os animais foram sacrificados e necropsiados. Os animais de ambos os tratamentos foram infectados por Salmonella e soroconverteram. Não houve diferença (P>0,05) entre os grupos nas médias de Salmonella, Enterococcus, Lactobacillus e coliformes totais nas fezes, porém a administração de probiótico resultou em menor frequência de isolamento de Salmonella a partir de fígado (P=0,04), linfonodos mesentéricos (P=0,04), pulmão (P=0,03) e baço (P=0,01). Conclui-se que os microrganismos probióticos testados não foram capazes de impedir a infecção ou a excreção fecal de Salmonella em suínos de crescimento, mas diminuíram o número de portadores em linfonodos mesentéricos.

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“…Standard OD values correspond to the average of triplicate medians obtained for negative and positive (weak, medium and strong) serum pools. The ELISA experimental cut-off point was set at the OD average obtained for the negative control serum samples plus four standard deviations, and was set at OD 0.169 (KICH et al, 2007). The %OD for serum was calculated following the equation, where NC is the optical density of a reference negative sample; and PC is the optical density of a reference strong positive sample.…”

Section: Methodsmentioning

confidence: 99%

“…Serum was used in the ELISA-Typhimurium test developed by Kich et al (2007), which is based on the phenol extracted LPS of S. Typhimurium (strain BRMSA 1084). Briefly, sera samples were diluted (1:400) in PBS pH 7.4 containing 0.05% Tween 20 (PBS-T) with 1% bovine serum albumin (PBS-TA) and added in triplicate to the plates coated with the antigen (1:2,000).…”

Section: Methodsmentioning

confidence: 99%

“…In Brazil, an in-house indirect ELISA test (ELISA-Typhimurium) based on the S. Typhimurium lipopolysaccharide (LPS) was developed (KICH et al, 2007) and proved to be able to detect sera antibodies against the most prevalent serovars of Salmonella found in the Brazilian swine production system. However, the cut-off initially established (experimental cut off) for the test presents limitations in the discrimination of herds with high and low number of positive pigs on Salmonella isolation.…”

Section: Introductionmentioning

confidence: 99%

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Assessment of different cut-off values of the ELISA-Typhimurium for the discrimination of swine herds with Salmonella isolation

Kich

Costa

Triques

et al. 2016

SCA

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Salmonella control programs require the classification of positive herds according with the prevalence of infected pigs. This must be based on diagnostic methods that are reliable, accurate and cost effective. Since serological methods fulfill most of these prerequisites, many countries have adopted indirect ELISA tests to discriminate herds, based on their seroprevalence. In view of that, an in-house indirect ELISA test (ELISA-Typhimurium) previously developed proved to be able to detect sera antibodies against the most prevalent serovars of Salmonella found in Brazilian pig farms. This study aimed to assess different cut-off values of the ELISA-Typhimurium in terms of their correlation with Salmonella isolation results at the herd level. Blood and mesenteric lymph node or rectal swabs were taken from 56 slaughter and 14 grower batches from different farms. Serum was tested using the ELISA-Typhimurium at different cut-off points (OD: 6.87%-40%). A total of 55 slaughter batches had at least one Salmonellapositive sample, and the within-herd prevalence was 64.9% (SE±24.8). All 14-grower batches were negative on Salmonella isolation. The logistic regression demonstrated that at 20% OD the ELISATyphimurium test is the best model to predict Salmonella isolation frequency at the herd level. At this cut-off point, the test may be a helpful tool for discriminating Salmonella infected herds.

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Development and Application of an Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Prevalent Salmonella Serovars in Swine in Southern Brazil

Cited by 20 publications

References 24 publications

Anti-LPS Test Strip for the Detection of Food Contaminated with Salmonella and E. coli

Anti-LPS Test Strip for the Detection of Food Contaminated with Salmonella and E. coli

Efeito de probiótico na infecção e excreção fecal de Salmonella em suínos

Assessment of different cut-off values of the ELISA-Typhimurium for the discrimination of swine herds with Salmonella isolation

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