{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:38:18Z","timestamp":1792787898579,"version":"4.5.0"},"reference-count":24,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,3,5]],"date-time":"2021-03-05T00:00:00Z","timestamp":1614902400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,3,5]],"date-time":"2021-03-05T00:00:00Z","timestamp":1614902400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the National Key Research and Development Program of China","award":["2016YFD0500905"],"award-info":[{"award-number":["2016YFD0500905"]}]},{"name":"the Modern Agricultural Industry Technology System Foundation of Shandong Province","award":["SDAIT-10-06"],"award-info":[{"award-number":["SDAIT-10-06"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Vet Res"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Klebsiella pneumonia<\/jats:italic>\n                      , a Gram-negative bacterium belonging to the genus\n                      <jats:italic>Enterobacter<\/jats:italic>\n                      , causes many human and livestock diseases. Notably, infected goats may develop pneumonia, septicemia, which can lead to occasional death, resulting in great economic losses in goat-farming industry. However, there are little systematic methods for detection of goat\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      in livestock production.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      In this study, we developed a\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      goat polyclonal antibody and established an indirect ELISA method to detect the\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      . After screening and optimizing the conditions for detection, we determined the optimal working dilutions of the coated-bacterial antigen, the polyclonal antibody, and the enzyme-labeled secondary antibody that were 1:800 (2.99\u2009\u00d7\u200910\n                      <jats:sup>7<\/jats:sup>\n                      CFU\/ml), 1:6400, and 1:5000, respectively. The optimal condition of coating and blocking were both 4\u2009\u00b0C for 12\u2009h. The optimal dilution buffers of bacterial antigen, the antibodies, and the blocking buffer were 0.05\u2009mol\/L carbonate buffer, 1% BSA phosphate buffer, and 1.5% BSA carbonate buffer, respectively. The cut-off value was determined to be 0.28, and the analytical sensitivity was 1:800 (dilution of a positive sample). Furthermore, there was no cross-reaction between the coated antigen and goat serum positive for antibodies against other bacteria, indicating that indirect ELISA could detect\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      specifically in most cases. The average coefficients of variation of intra-assay and inter-assay were 4.37 and 5.17% indicating favorable reproducibility of indirect ELISA. In the detection of clinical veterinary samples, the positive rate of indirect ELISA was 6.74%, higher than that of conventional agglutination assays.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      Taken together, we successfully established an indirect ELISA method for detecting antibodies against\n                      <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                      in goats, which can be applied in production.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12917-021-02820-1","type":"journal-article","created":{"date-parts":[[2021,3,4]],"date-time":"2021-03-04T22:02:32Z","timestamp":1614895352000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Establishment and evaluation of an indirect ELISA for detection of antibodies to goat Klebsiella pneumonia"],"prefix":"10.1186","volume":"17","author":[{"given":"Ruichang","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongqi","family":"Shang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiangyun","family":"Niu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongqiang","family":"Miao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhou","family":"Sha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liting","family":"Qin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Duo","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruiliang","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,3,5]]},"reference":[{"key":"2820_CR1","doi-asserted-by":"crossref","unstructured":"Kamal G, Rahul R, Mandira V-B, Ashok S. 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