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In this study, we developed sandwich ELISA with detection range of 10<jats:sup>2<\/jats:sup> to 10<jats:sup>8<\/jats:sup> cells mL<jats:sup>\u22121<\/jats:sup> and limit of detection at 10<jats:sup>3<\/jats:sup> cells mL<jats:sup>\u22121<\/jats:sup> by employing polyclonal rabbit IgG (capture antibody, 10\u2009\u00b5g\u2009mL<jats:sup>\u22121<\/jats:sup>) and mice IgG (detection antibody, 50\u2009\u00b5g\u2009mL<jats:sup>\u22121<\/jats:sup>) antibody for its detection. Surface Plasmon Resonance evaluated the interaction of detection antibody with whole cell spiked serum samples at LOD of 10<jats:sup>2<\/jats:sup> cells mL<jats:sup>\u22121<\/jats:sup> along with non co-operative interaction of protein albumin. Further, kinetic evaluation study using detection antibody against cell envelope antigen was performed whereby, Equilibrium Dissociation Constant (K<jats:sub>D<\/jats:sub>) and Maximum Binding Capacity (B<jats:sub>max<\/jats:sub>) were found to be 16.48 pM and 81.67\u2009m\u00b0 for <jats:italic>Brucella abortus<\/jats:italic> S99 and 0.42 pM and 54.50\u2009m\u00b0 for <jats:italic>Brucella melitensis<\/jats:italic> 16\u2009M, respectively. During interference study, sandwich ELISA assay cross-reacted with either of the polyclonal antibody of above <jats:italic>Brucella<\/jats:italic> species. Upon validation, no cross-reactivity observed with bacteria-closely related to <jats:italic>Brucella<\/jats:italic>. In conclusion, developed semi-quantitative sandwich immunoassay is sensitively rapid in whole cell detection of <jats:italic>Brucella<\/jats:italic> and will be useful in development of detection assays from environmental and clinical matrices.<\/jats:p>","DOI":"10.1038\/s41598-020-65347-9","type":"journal-article","created":{"date-parts":[[2020,5,22]],"date-time":"2020-05-22T10:04:00Z","timestamp":1590141840000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Development and validation of immunoassay for whole cell detection of Brucella abortus and Brucella melitensis"],"prefix":"10.1038","volume":"10","author":[{"given":"Richa","family":"Hans","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4398-9838","authenticated-orcid":false,"given":"Pranjal Kumar","family":"Yadav","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pushpendra Kumar","family":"Sharma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mannan","family":"Boopathi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Duraipandian","family":"Thavaselvam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,5,22]]},"reference":[{"key":"65347_CR1","first-page":"1511","volume":"7","author":"S Umesha","year":"2018","unstructured":"Umesha, S. & Raghava, S. 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