{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T02:07:16Z","timestamp":1792807636261,"version":"4.5.0"},"reference-count":43,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T00:00:00Z","timestamp":1755043200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T00:00:00Z","timestamp":1755043200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"Medical Science and Technology Project","award":["JK2023gk002"],"award-info":[{"award-number":["JK2023gk002"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2025,8,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Yersinia pestis<\/jats:italic>\n                      is the causative agent of plague, a human disease with potentially devastating consequences. Here, we developed an enzyme-linked immunosorbent assay-like visual detection method based on clustered regularly interspaced short palindromic repeats (CRISPR) detection and DNAzyme for the cost-effective and highly sensitive detection of\n                      <jats:italic toggle=\"yes\">Y. pestis<\/jats:italic>\n                      . A novel specific gene sequence (CH57_3927) was screened for the detection target of\n                      <jats:italic toggle=\"yes\">Y. pestis<\/jats:italic>\n                      . The recombinase-aided amplification (RAA) assay, CRISPR\/Cas12a detection assay, and G-quadruplex (G4) DNAzyme-based color development assay were separately established and optimized. These three optimized assays were integrated into an advanced ELISA-like visual detection method\u2014RAA-CRISPR\/Cas12a-DNAzyme (RCCD)\u2014by further optimization of their components to improve the compatibility between them. The amplified target sequence binds to crRNA and activates the Cas12a nucleases for trans-cleave G4. As a result, the cleaved G4 is unable to bind with hemin to exert peroxidase activity, thus impeding the catalysis of the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS\n                      <jats:sup>2\u2013<\/jats:sup>\n                      ) colorimetric reaction. Consequently, negative samples exhibit a dark green coloration, while the positive products appear nearly colorless, facilitating visual differentiation with the naked eye. In addition, the RCCD detection platform effectively distinguished\n                      <jats:italic toggle=\"yes\">Y. pestis<\/jats:italic>\n                      from all other closely related species, with a detection limit of 1 copy\/reaction. Evaluated using\n                      <jats:italic toggle=\"yes\">Y. pestis<\/jats:italic>\n                      DNA-spiked blood samples and uninfected samples, both sensitivity and specificity were 100%. The method shows significant potential for detecting targets in clinical samples and is well-suited for use in resource-limited environments. It offers advantages such as visual detection, batch detection, and low cost.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        We utilized Mauve software to screen\n                        <jats:italic toggle=\"yes\">Yersinia pestis<\/jats:italic>\n                        specific genes and integrated CRISPR-Cas12a, RAA amplification, and G-quadruplex DNAzyme technology to establish an advanced ELISA-like visual detection method. The visual detection method offers a more cost-effective alternative compared to the conventional CRISPR detection method that relies on fluorescence-labeled ssDNA reporter or lateral flow (LF) test strips. With only one thermostatic device required, it enhances the convenience of rapid on-site screening of\n                        <jats:italic toggle=\"yes\">Y. pestis<\/jats:italic>\n                        outbreaks, providing effective support for plague detection, prevention, and control within primary medical and health institutions.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jcm.00274-25","type":"journal-article","created":{"date-parts":[[2025,7,24]],"date-time":"2025-07-24T13:00:52Z","timestamp":1753362052000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":4,"title":["An ELISA-like sensitive and visual detection system targeting\n                    <i>Yersinia pestis<\/i>\n                    based on CRISPR\/Cas12a and DNAzyme"],"prefix":"10.1128","volume":"63","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-1133-0052","authenticated-orcid":true,"given":"Yingqing","family":"Mao","sequence":"first","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruichen","family":"Lv","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Shao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00hj8s172","id-type":"ROR","asserted-by":"publisher"}],"name":"Teachers College, Columbia University","place":["Manhattan, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhu","family":"Wang","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiong","family":"Chen","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiming","family":"Yi","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yixin","family":"Ge","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongming","family":"Wang","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8491-4402","authenticated-orcid":false,"given":"Yuexi","family":"Li","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6186-9023","authenticated-orcid":true,"given":"Yong","family":"Qi","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques","place":["Nanjing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0802.010164"},{"key":"e_1_3_3_3_2","first-page":"40","article-title":"Human plague: review of regional morbidity and mortality, 2004-2009","volume":"6","year":"2009","unstructured":"Human plague: review of regional morbidity and mortality, 2004-2009. 2009. 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