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Then, a specific extracellular region of a transmembrane protein with the potential for diagnosis was identified. After that, a monoclonal antibody (Mab) named 3A8 was obtained using this extracellular region as an immunogen. Finally, a cELISA was established with the extracellular domain of this transmembrane protein as the coating antigen, Mab 3A8 as the competitive antibody, and HRP-labeled goat anti-mouse IgG as the enzyme-labeled antibody. This established method was used to detect the antibody dynamic regularity of goats which are artificially immunized <jats:italic>Mmm<\/jats:italic> and was also compared with a commercial ELISA kit. Further, the sera of 1011 different cattle from border provinces of China were monitored using a candidate Mab 3A8 cELISA. The detection results of known background sera used in this study indicate that a candidate diagnostic marker was successfully identified by analyzing all the coding proteins of <jats:italic>Mmm<\/jats:italic> in this research, and the cELISA established based on the Mab 3A8 against this protein can detect CBPP-positive serum with specificity and has no cross-reaction with other related epidemic disease-positive sera. In addition, we tested the sera collected from the border areas of China using the established ELISA, and no positive sample was detected. The research protocol of the CBPP cELISA established in this study is different from the traditional method, which can greatly reduce the investment of manpower and capital and save development time. We believe that this study\u2019s protocol could serve as a reference for the development of detection methods for mycoplasma and other complex pathogens.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\u2022 <jats:italic>A Mmm-specific diagnostic marker was obtained based on protein characteristics.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022 <jats:italic>A cELISA was established for CBPP serum antibody detection.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\u2022 <jats:italic>The serological investigation was conducted for CBPP in the border areas of China.<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00253-024-13127-0","type":"journal-article","created":{"date-parts":[[2024,4,8]],"date-time":"2024-04-08T10:02:05Z","timestamp":1712570525000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A candidate competitive ELISA based on monoclonal antibody 3A8 for diagnosis of contagious bovine pleuropneumonia"],"prefix":"10.1007","volume":"108","author":[{"given":"Qi","family":"Wu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhixin","family":"Ma","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiao","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tong","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yidan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiuqing","family":"Xin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0393-8726","authenticated-orcid":false,"given":"Qingyuan","family":"Xu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,4,8]]},"reference":[{"issue":"6","key":"13127_CR1","doi-asserted-by":"publisher","first-page":"e00945","DOI":"10.1128\/iai.00945-19","volume":"88","author":"JY Adamu","year":"2020","unstructured":"Adamu JY, Wawegama NK, Condello AK, Marenda MS, Markham PF, Browning GF, Tivendale KA (2020) Mycoplasma bovis membrane protein MilA is a multifunctional lipase with novel lipid and glycosaminoglycan binding activity. 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The protocols were approved by the Committee on the Ethics of Animal Experiments of the Harbin Veterinary Research lnstitute (HVRI) of the Chinese Academy of Agricultural Sciences (CAAS).","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}],"article-number":"290"}}