{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T05:04:49Z","timestamp":1789621489806,"version":"build-2803163510"},"reference-count":44,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Respiratory disease caused by bacterial and mycoplasmal pathogens is an important threat to goat health, and mixed infections complicate etiological diagnosis. Rapid multiplex detection is therefore valuable for clinical investigation and surveillance.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Two triplex TaqMan real-time quantitative PCR (qPCR) panels were established for the simultaneous detection of six major caprine respiratory pathogens. Panel I targeted\n                      <jats:italic>Mycoplasma ovipneumoniae<\/jats:italic>\n                      (Mo),\n                      <jats:italic>Mycoplasma mycoides<\/jats:italic>\n                      subsp.\n                      <jats:italic>capri<\/jats:italic>\n                      (Mmc), and\n                      <jats:italic>Mycoplasma capricolum<\/jats:italic>\n                      subsp.\n                      <jats:italic>capripneumoniae<\/jats:italic>\n                      (Mccp), whereas Panel II targeted\n                      <jats:italic>Pasteurella multocida<\/jats:italic>\n                      (Pm),\n                      <jats:italic>Mannheimia haemolytica<\/jats:italic>\n                      (Mh), and\n                      <jats:italic>Streptococcus equi<\/jats:italic>\n                      subsp.\n                      <jats:italic>zooepidemicus<\/jats:italic>\n                      (Sz). Analytical specificity was assessed using target plasmids and nucleic acids from target and non-target pathogens, whereas analytical sensitivity and reproducibility were evaluated using recombinant plasmid standards. Clinical applicability was assessed separately using 200 field-collected goat nasal swab samples.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      No amplification was observed for the tested non-target organisms. The limits of detection were 1.73\u202f\u00d7\u202f10\n                      <jats:sup>2<\/jats:sup>\n                      copies\/\u03bcL for Mo, 2.01\u202f\u00d7\u202f10\n                      <jats:sup>3<\/jats:sup>\n                      copies\/\u03bcL for Mmc, 3.63\u202f\u00d7\u202f10\n                      <jats:sup>2<\/jats:sup>\n                      copies\/\u03bcL for Mccp, 1.03\u202f\u00d7\u202f10\n                      <jats:sup>2<\/jats:sup>\n                      copies\/\u03bcL for Pm, 1.59\u202f\u00d7\u202f10\n                      <jats:sup>2<\/jats:sup>\n                      copies\/\u03bcL for Mh, and 5.88\u202f\u00d7\u202f10\n                      <jats:sup>2<\/jats:sup>\n                      copies\/\u03bcL for Sz. Intra-assay CVs ranged from 0.55 to 2.76%, and inter-assay CVs ranged from 0.38 to 3.10%. Multiplex qPCR detected at least one target in 117 of 200 clinical samples (58.5%), including 47 samples with two or more targets (23.5%).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The multiplex qPCR assay developed in this study provides a rapid and specific tool for the simultaneous detection of six major respiratory pathogens in goats, with potential utility for clinical diagnosis and epidemiological surveillance.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2026.1925484","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T04:47:00Z","timestamp":1789620420000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and validation of a multiplex TaqMan real-time quantitative PCR assay for the simultaneous detection of six major bacterial respiratory pathogens in goats"],"prefix":"10.3389","volume":"13","author":[{"given":"Mingzhe","family":"Fu","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Yangling, Shaanxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Yangling, Shaanxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeyang","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Yangling, Shaanxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weimin","family":"Li","sequence":"additional","affiliation":[{"name":"Datong District Animal Husbandry Comprehensive Service Center","place":["Daqing, Heilongjiang, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Yangling, Shaanxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingang","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Northwest A&F University","place":["Yangling, Shaanxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1016\/S0928-4249(98)80026-6","article-title":"Pasteurella haemolytica complicated respiratory infections in sheep and goats","volume":"29","author":"Brogden","year":"1998","journal-title":"Vet 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