An optimized double-antibody sandwich ELISA for quantitative detection of WSSV in artificially infected crayfish

Porcine deltacoronavirus (PDCoV) can cause diarrhea and dehydration in newborn piglets. Here, we developed a double antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-ELISA) for detection of PDCoV by using a specific monoclonal antibody against the PDCoV N protein and an anti-PDCoV rabbit polyclonal antibody. Using DAS-ELISA, the detection limit of recombinant PDCoV N protein and virus titer were approximately 0.5 ng/mL and 103.0 TCID50/mL, respectively. A total of 59 intestinal and 205 fecal samples were screened for the presence of PDCoV by using DAS-ELISA and reverse transcriptase real-time PCR (RT-qPCR). The coincidence rate of the DAS-ELISA and RT-qPCR was 89.8%. DAS-ELISA had a sensitivity of 80.8% and specificity of 95.6%. More importantly, the DAS-ELISA could detect the antigen of PDCoV inactivated virus, and the viral antigen concentrations remained unchanged in the inactivated virus. These results suggest that DAS-ELISA could be used for antigen detection of clinical samples and inactivated vaccines. It is a novel method for detecting PDCoV infections and evaluating the PDCoV vaccine.

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Development and evaluation of indirect double-antibody sandwich ELISA for rapid detection of Salmonid Alphavirus using Baculoviridae expressed E1 Protein

Aquaculture ◽

10.1016/j.aquaculture.2021.737081 ◽

2021 ◽

pp. 737081

Author(s):

Shuai Gao ◽

Na Wang ◽

Jiawei Yang ◽

Jinhui Sun ◽

Yuting Wang ◽

...

Keyword(s):

Rapid Detection ◽

Sandwich Elisa ◽

Double Antibody ◽

E1 Protein

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Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food

Scientific Reports ◽

10.1038/srep16092 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 34

Author(s):

Longjiao Zhu ◽

Jing He ◽

Xiaohan Cao ◽

Kunlun Huang ◽

Yunbo Luo ◽

...

Keyword(s):

Bacillus Cereus ◽

Rapid Detection ◽

Sandwich Elisa ◽

Double Antibody

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Evaluation of indirect competitive and double antibody sandwich ELISA tests to determine β-lactoglobulin and ovomucoid in model processed foods

Food and Agricultural Immunology ◽

10.1080/09540100802520755 ◽

2008 ◽

Vol 19 (4) ◽

pp. 339-350 ◽

Cited By ~ 13

Author(s):

Ruth de Luis ◽

Luis Mata ◽

Gloria Estopañán ◽

María Lavilla ◽

Lourdes Sánchez ◽

...

Keyword(s):

Sandwich Elisa ◽

Processed Foods ◽

Double Antibody ◽

Β Lactoglobulin

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Quantitative Detection of Horse Contamination in Cooked Meat Products by ELISA

Journal of AOAC International ◽

10.5740/jaoacint.17-0151 ◽

2018 ◽

Vol 101 (3) ◽

pp. 817-823 ◽

Cited By ~ 3

Author(s):

Cortlandt P Thienes ◽

Jongkit Masiri ◽

Lora A Benoit ◽

Brianda Barrios-Lopez ◽

Santosh A Samuel ◽

...

Keyword(s):

Sandwich Elisa ◽

Meat Products ◽

Rapid Test ◽

Cross Reactivity ◽

Quantitative Detection ◽

Analytical Sensitivity ◽

Horse Meat ◽

Analytical Range ◽

Cooked Meat ◽

Cooked Meat Products

Abstract Concerns about the contamination of meat products with horse meat and new regulations for the declaration of meat adulterants have highlighted the need for a rapid test to detect horse meat adulteration. To address this need, Microbiologique, Inc., has developed a sandwich ELISA that can quantify the presence of horse meat down to 0.1% (w/w) in cooked pork, beef, chicken, goat, and lamb meats. This horse meat authentication ELISA has an analytical sensitivity of 0.000030 and 0.000046% (w/v) for cooked and autoclaved horse meat, respectively, and an analytical range of quantitation of 0.05–0.8% (w/v) in the absence of other meats. The assay is rapid and can be completed in 1 h and 10 min. Moreover, the assay is specific for cooked horse meat and does not demonstrate any cross-reactivity with xenogeneic cooked meat sources.

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