Development of double-antibody sandwich ELISA for rapidly quantitative detection of antigen concentration in inactivated SCRV vaccine

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 of a Double-Antibody Sandwich ELISA for Rapid Detection of the MCP Antigen Concentration in Inactivated ISKNV Vaccines

Vaccines ◽

10.3390/vaccines9111264 ◽

2021 ◽

Vol 9 (11) ◽

pp. 1264

Author(s):

Hongru Liang ◽

Lixi Zhang ◽

Xiaozhe Fu ◽

Qiang Lin ◽

Lihui Liu ◽

...

Keyword(s):

Sandwich Elisa ◽

High Morbidity ◽

Antigen Concentration ◽

Assay Sensitivity ◽

Effective Prevention ◽

Siniperca Chuatsi ◽

Standard Curve ◽

Double Antibody ◽

Inactivated Vaccines ◽

And Control

Infectious spleen and kidney necrosis virus (ISKNV) resulted in severe systemic diseases with high morbidity and mortality in Siniperca chuatsi. Vaccination is the primary method for effective prevention and control of these diseases. The development of inactivated ISKNV vaccines made some progress, but the technique of quality evaluation is scarce. Herein, a measurement of the MCP (major capsid protein) antigen concentration for the inactivated ISKNV vaccine was developed by double-antibody sandwich ELISA. Firstly, mouse monoclonal antibodies against ISKNV particles and MCP were generated. Then, a double-antibody sandwich ELISA was developed using the monoclonal antibody 1C8 1B9 as the capture antibody and Biotin-3B12 6B3 as the detection antibody. A standard curve was generated using the MCP concentration versus OD value with the linear range of concentration of 4.69~300 ng/mL. The assay sensitivity was 0.9 ng/mL. The antigen content of three batches of inactivated ISKNV vaccines was quantitatively detected using the double-antibody sandwich ELISA. The results showed that MCP antigen contents of inactivated ISKNV vaccines were positively correlated with the viral titers. The newly established double-antibody sandwich ELISA provided a useful tool for the detection of antigen quality for ISKNV inactivated vaccines.

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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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