{"title":"基于重组病毒蛋白2的酶联免疫吸附法检测传染性法氏囊病病毒抗体的建立","authors":"Leticia Keller, Carina Romanutti, Flavia Adriana Zanetti","doi":"10.1016/j.ram.2024.09.003","DOIUrl":null,"url":null,"abstract":"<div><div>Infectious bursal disease virus (IBDV) is the etiological agent of a highly contagious and immunosuppressive disease in chickens. In poultry farms, the level of anti-IBDV antibodies of numerous serum samples must be monitored using fast and simple methodologies. Therefore, the aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA), in an indirect format, using a version of mature viral protein 2 (VP2) of IBDV as coating agent. This recombinant fusion protein (His-VP2) was expressed at high levels in <em>Escherichia coli</em>. Bacterial inclusion bodies containing His-VP2 were successfully recovered using a simple, inexpensive and efficient method, a further purification of recombinant protein by affinity chromatography using immobilized metal chelates being unnecessary. After the VP2-ELISA was optimized, its performance was evaluated using preanalyzed sera from uninfected specific pathogen-free chickens and broilers vaccinated against IBDV in poultry farms, using a commercial ELISA kit. Based on these results, the developed assay proved to be sensitive, specific and in high agreement with the kit available on the market. In addition, the in-house ELISA demonstrated to be reproducible by intra-assay and inter-assay variability studies. In conclusion, VP2-ELISA could be an efficient and low-cost alternative diagnostic method to detect antibodies to IBDV in the poultry industry.</div></div>","PeriodicalId":21163,"journal":{"name":"Revista Argentina de microbiologia","volume":"57 3","pages":"Pages 208-216"},"PeriodicalIF":2.1000,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of an enzyme-linked immunosorbent assay for detection of antibodies against infectious bursal disease virus based on a version of the recombinant viral protein 2\",\"authors\":\"Leticia Keller, Carina Romanutti, Flavia Adriana Zanetti\",\"doi\":\"10.1016/j.ram.2024.09.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Infectious bursal disease virus (IBDV) is the etiological agent of a highly contagious and immunosuppressive disease in chickens. In poultry farms, the level of anti-IBDV antibodies of numerous serum samples must be monitored using fast and simple methodologies. Therefore, the aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA), in an indirect format, using a version of mature viral protein 2 (VP2) of IBDV as coating agent. This recombinant fusion protein (His-VP2) was expressed at high levels in <em>Escherichia coli</em>. Bacterial inclusion bodies containing His-VP2 were successfully recovered using a simple, inexpensive and efficient method, a further purification of recombinant protein by affinity chromatography using immobilized metal chelates being unnecessary. After the VP2-ELISA was optimized, its performance was evaluated using preanalyzed sera from uninfected specific pathogen-free chickens and broilers vaccinated against IBDV in poultry farms, using a commercial ELISA kit. Based on these results, the developed assay proved to be sensitive, specific and in high agreement with the kit available on the market. In addition, the in-house ELISA demonstrated to be reproducible by intra-assay and inter-assay variability studies. In conclusion, VP2-ELISA could be an efficient and low-cost alternative diagnostic method to detect antibodies to IBDV in the poultry industry.</div></div>\",\"PeriodicalId\":21163,\"journal\":{\"name\":\"Revista Argentina de microbiologia\",\"volume\":\"57 3\",\"pages\":\"Pages 208-216\"},\"PeriodicalIF\":2.1000,\"publicationDate\":\"2025-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Revista Argentina de microbiologia\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0325754124001160\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Revista Argentina de microbiologia","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0325754124001160","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Development of an enzyme-linked immunosorbent assay for detection of antibodies against infectious bursal disease virus based on a version of the recombinant viral protein 2
Infectious bursal disease virus (IBDV) is the etiological agent of a highly contagious and immunosuppressive disease in chickens. In poultry farms, the level of anti-IBDV antibodies of numerous serum samples must be monitored using fast and simple methodologies. Therefore, the aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA), in an indirect format, using a version of mature viral protein 2 (VP2) of IBDV as coating agent. This recombinant fusion protein (His-VP2) was expressed at high levels in Escherichia coli. Bacterial inclusion bodies containing His-VP2 were successfully recovered using a simple, inexpensive and efficient method, a further purification of recombinant protein by affinity chromatography using immobilized metal chelates being unnecessary. After the VP2-ELISA was optimized, its performance was evaluated using preanalyzed sera from uninfected specific pathogen-free chickens and broilers vaccinated against IBDV in poultry farms, using a commercial ELISA kit. Based on these results, the developed assay proved to be sensitive, specific and in high agreement with the kit available on the market. In addition, the in-house ELISA demonstrated to be reproducible by intra-assay and inter-assay variability studies. In conclusion, VP2-ELISA could be an efficient and low-cost alternative diagnostic method to detect antibodies to IBDV in the poultry industry.
期刊介绍:
La Revista Argentina de Microbiología es una publicación trimestral editada por la Asociación Argentina de Microbiología y destinada a la difusión de trabajos científicos en las distintas áreas de la Microbiología. La Asociación Argentina de Microbiología se reserva los derechos de propiedad y reproducción del material aceptado y publicado.