{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:27:26Z","timestamp":1792787246592,"version":"4.5.0"},"reference-count":25,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32160847"],"award-info":[{"award-number":["32160847"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004317","name":"Shihezi University","doi-asserted-by":"publisher","award":["CGZH202309"],"award-info":[{"award-number":["CGZH202309"]}],"id":[{"id":"10.13039\/501100004317","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the 12th Division of Xinjiang Production and Construction Corps","award":["SRS2022013"],"award-info":[{"award-number":["SRS2022013"]}],"id":[{"id":"https:\/\/ror.org\/03hcmxw73","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      A combined molecular strategy was developed to accurately differentiate wild-type and vaccine strains of\n                      <jats:italic toggle=\"yes\">Bacillus anthracis<\/jats:italic>\n                      . This approach integrates a multiplex TaqMan qPCR assay targeting the\n                      <jats:italic toggle=\"yes\">BclA<\/jats:italic>\n                      ,\n                      <jats:italic toggle=\"yes\">sap<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">cap<\/jats:italic>\n                      genes with conventional PCR analysis of\n                      <jats:italic toggle=\"yes\">BclA<\/jats:italic>\n                      fragment length polymorphism. The qPCR assay demonstrated high specificity, with no cross-reactivity against other common pathogens, and exhibited a consistent limit of detection of 2.11 \u00d7 10\n                      <jats:sup>1<\/jats:sup>\n                      copies\/\u03bcL (Cq &lt; 35) for all three targets. Excellent repeatability was confirmed, with intra- and inter-assay coefficients of variation below 1.96% and 3.06%, respectively. Validation using 19 wild-type strains revealed two distinct qPCR profiles, and all wild-type strains were conclusively differentiated from the capsule-producing Chinese vaccine strain II and the acapsular Sterne strain by their characteristic\n                      <jats:italic toggle=\"yes\">BclA<\/jats:italic>\n                      amplicon sizes. Through simulation PCR validation of common global canSNP genotyping (groups A, B, and C), it is possible to distinguish between wild-type and vaccine strains in China. This integrated \u201cmultiplex qPCR\u2013BclA length analysis\u201d protocol provides a reliable, sensitive, and specific tool for the precise detection and differentiation of\n                      <jats:italic toggle=\"yes\">B. anthracis<\/jats:italic>\n                      strains in China.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Anthrax, which is caused by the bacterium\n                        <jats:italic toggle=\"yes\">B. anthracis<\/jats:italic>\n                        , remains a serious zoonotic disease. Its effective surveillance relies on precise strain identification. Currently, it is challenging to rapidly and accurately distinguish wild-type strains from vaccine strains, which hinders outbreak investigation and response. This study develops a novel molecular strategy combining multiple genetic targets to solve this critical diagnostic problem. The new method provides a reliable tool for strain differentiation, directly supporting accurate source tracing and informed decision-making during anthrax outbreaks. By improving diagnostic precision, this work contributes to strengthening public and veterinary health defenses against this persistent threat.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.00192-26","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T13:00:11Z","timestamp":1788267611000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["An integrated strategy of multiplex qPCR and\n                    <i>BclA<\/i>\n                    gene length polymorphism analysis for differentiating\n                    <i>Bacillus anthracis<\/i>\n                    wild-type and vaccine strains"],"prefix":"10.1128","volume":"14","author":[{"given":"Jiaqi","family":"Yang","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology, Shihezi University","place":["Shihezi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Liang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Shihezi University","place":["Shihezi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Liang","sequence":"additional","affiliation":[{"name":"Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanfang","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Shihezi University","place":["Shihezi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9514-6760","authenticated-orcid":false,"given":"Lingwei","family":"Zhu","sequence":"additional","affiliation":[{"name":"Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences","place":["Changchun, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2649-4144","authenticated-orcid":true,"given":"Yonggang","family":"Qu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology, Shihezi University","place":["Shihezi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","volume-title":"Anthrax in humans and animals","author":"WHO Guidelines Approved by the Guidelines Review Committee","year":"2008","unstructured":"WHO Guidelines Approved by the Guidelines Review Committee. 2008. Anthrax in humans and animals. World Health Organization Copyright \u00a9 World Health Organization 2008."},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-019-0435-4"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2000.03000.x"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.5.1555-1563.2003"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.718171"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2025.117279"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/CVI.00023-06"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2014.05.008"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2004.10.005"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2008.06.001"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11274-010-0592-3"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.10.2928-2936.2000"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0122004"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-024-09817-9"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01501-16"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000461"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.2478\/jvetres-2018-0075"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-010-2848-0"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1007\/s12223-021-00869-x"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biologicals.2020.11.004"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2005.04.032"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01069-09"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0006908"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-020-4867-5"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micres.2020.126414"}],"container-title":["Microbiology Spectrum"],"original-title":[],"contributor":[{"given":"Dongshu","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"reviewer"}]}],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00192-26","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.00192-26","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T14:51:53Z","timestamp":1792507913000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.00192-26"}},"subtitle":[],"editor":[{"given":"Cecilia M.","family":"Thompson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,10,6]]},"references-count":25,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2026,10,6]]}},"alternative-id":["10.1128\/spectrum.00192-26"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.00192-26","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,6]]},"assertion":[{"value":"2026-01-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-24","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e00192-26"}}