{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T19:53:01Z","timestamp":1790970781390,"version":"4.1.0"},"update-to":[{"DOI":"10.1371\/journal.pntd.0011596","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T00:00:00Z","timestamp":1694476800000}}],"reference-count":26,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2023,8,25]],"date-time":"2023-08-25T00:00:00Z","timestamp":1692921600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Second Century Fund"},{"name":"Center of Excellence in Critical Care Nephrology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand."},{"name":"The Tropical Medicine Cluster, Chulalongkorn University."}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Lack of available sensitive point-of-care testing is one of the primary obstacles to the rapid diagnosis of leptospirosis. The purpose of this study was to test the performance of two point-of-care tests, a clustered regularly interspaced short palindromic repeat (CRISPR)\/CRISPR-associated protein 12a (CRISPR\/Cas12a) fluorescence-based diagnostic assay (FBDA), a\n                      <jats:italic>Leptospira<\/jats:italic>\n                      immunoglobulin M (IgM) rapid diagnostic test (RDT), and the two tests combined.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methodology\/Principal findings<\/jats:title>\n                    <jats:p>\n                      For the diagnosis of 171 clinical samples, a recombinase polymerase amplification (RPA)-CRISPR\/Cas12a FBDA for whole blood and\n                      <jats:italic>Leptospira<\/jats:italic>\n                      IgM RDT (Medical Science Public Health, Thailand) for serum were used. The confirmed cases were determined by using any positive qPCR, microscopic agglutination test (MAT), and culture results. Diagnostic accuracy was assessed on the first day of enrollment and stratified by the day after symptom onset. The overall sensitivity of the\n                      <jats:italic>Leptospira<\/jats:italic>\n                      IgM RDT and RPA-CRISPR\/Cas12a FBDA was 55.66% and 60.38%, respectively. When the two tests were combined, the sensitivity rose to 84.91%. The specificity of each test was 63.08% and 100%, respectively, and 63.08% when combined. The sensitivity of the\n                      <jats:italic>Leptospira<\/jats:italic>\n                      IgM RDT rose on days 4\u20136 after the onset of fever, while the RPA-CRISPR\/Cas12a FBDA continued to decrease. When the two tests were combined, the sensitivity was over 80% at different days post-onset of fever.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Conclusions\/Significance<\/jats:title>\n                    <jats:p>\n                      The combination of\n                      <jats:italic>Leptospira<\/jats:italic>\n                      IgM RDT and RPA-CRISPR\/Cas12 FBDA exhibited significant sensitivity for the detection of leptospires at various days after the onset of fever, thereby reducing the likelihood of misdiagnosis. The combination of these assays may be suitable for early leptospirosis screening in situations with limited resources.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pntd.0011596","type":"journal-article","created":{"date-parts":[[2023,8,25]],"date-time":"2023-08-25T13:35:48Z","timestamp":1692970548000},"page":"e0011596","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["The combination of RPA-CRISPR\/Cas12a and Leptospira IgM RDT enhances the early detection of leptospirosis"],"prefix":"10.1371","volume":"17","author":[{"given":"Sirawit","family":"Jirawannaporn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umaporn","family":"Limothai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sasipha","family":"Tachaboon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janejira","family":"Dinhuzen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patcharakorn","family":"Kiatamornrak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Watchadaporn","family":"Chaisuriyong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8544-8132","authenticated-orcid":true,"given":"Nattachai","family":"Srisawat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,8,25]]},"reference":[{"issue":"9","key":"pntd.0011596.ref001","doi-asserted-by":"crossref","first-page":"e0003898","DOI":"10.1371\/journal.pntd.0003898","article-title":"Global Morbidity and Mortality of Leptospirosis: A Systematic Review.","volume":"9","author":"F Costa","year":"2015","journal-title":"PLoS Negl Trop Dis"},{"issue":"2","key":"pntd.0011596.ref002","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1128\/CMR.14.2.296-326.2001","article-title":"Leptospirosis.","volume":"14","author":"PN Levett","year":"2001","journal-title":"Clin Microbiol Rev"},{"key":"pntd.0011596.ref003","first-page":"105183","article-title":"Leptospirosis: Increasing importance in developing countries","volume":"201","author":"ZMP Soo","year":"2019","journal-title":"Acta Trop"},{"issue":"6","key":"pntd.0011596.ref004","doi-asserted-by":"crossref","first-page":"2154","DOI":"10.1128\/JCM.02452-10","article-title":"Comparison of Real-Time PCR Assays for Detection of Pathogenic Leptospira spp. in Blood and Identification of Variations in Target Sequences","volume":"49","author":"P Bourhy","year":"2011","journal-title":"J Clin Microbiol"},{"issue":"1","key":"pntd.0011596.ref005","first-page":"199","article-title":"Leptospirosis Diagnosis: Competancy of Various Laboratory Tests","volume":"8","author":"SV Budihal","year":"2014","journal-title":"Journal of Clinical and Diagnostic Research"},{"issue":"4","key":"pntd.0011596.ref006","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.jmii.2013.03.001","article-title":"Laboratory diagnosis of leptospirosis: a challenge.","volume":"46","author":"D Musso","year":"2013","journal-title":"Journal of Microbiology, Immunology and Infection."},{"issue":"3","key":"pntd.0011596.ref007","doi-asserted-by":"crossref","first-page":"e0006319","DOI":"10.1371\/journal.pntd.0006319","article-title":"Thai-Lepto-on-admission probability (THAI-LEPTO) score as an early tool for initial diagnosis of leptospirosis: Result from Thai-Lepto AKI study group.","volume":"12","author":"T Sukmark","year":"2018","journal-title":"PLoS Negl Trop Dis."},{"key":"pntd.0011596.ref008","volume-title":"surveillance and control","author":"World Health Organization","year":"2003"},{"issue":"3","key":"pntd.0011596.ref009","first-page":"195","article-title":"Laboratory diagnosis of leptospirosis.","volume":"51","author":"SN Ahmad","year":"2005","journal-title":"J Postgrad Med"},{"issue":"1","key":"pntd.0011596.ref010","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.medmal.2012.11.005","article-title":"Diagnosisand epidemiology of leptospirosis","volume":"43","author":"M. Picardeau","year":"2013","journal-title":"Med Mal Infect"},{"issue":"4","key":"pntd.0011596.ref011","doi-asserted-by":"crossref","first-page":"395","DOI":"10.3390\/pathogens11040395","article-title":"Role of Diagnostics in Epidemiology, Management, Surveillance, and Control of Leptospirosis.","volume":"11","author":"JE Sykes","year":"2022","journal-title":"Pathogens."},{"issue":"1","key":"pntd.0011596.ref012","doi-asserted-by":"crossref","first-page":"3215","DOI":"10.1038\/s41598-021-82290-5","article-title":"Human, animal, water source interactions and leptospirosis in Thailand.","volume":"11","author":"U Narkkul","year":"2021","journal-title":"Sci Rep"},{"issue":"2","key":"pntd.0011596.ref013","doi-asserted-by":"crossref","first-page":"e0009159","DOI":"10.1371\/journal.pntd.0009159","article-title":"A prospective study to evaluate the accuracy of rapid diagnostic tests for diagnosis of human leptospirosis: Result from THAI-LEPTO AKI study.","volume":"15","author":"J Dinhuzen","year":"2021","journal-title":"PLoS Negl Trop Dis."},{"issue":"1","key":"pntd.0011596.ref014","doi-asserted-by":"crossref","first-page":"e0010112","DOI":"10.1371\/journal.pntd.0010112","article-title":"Rapid and sensitive point-of-care detection of Leptospira by RPA-CRISPR\/Cas12a targeting lipL32.","volume":"16","author":"S Jirawannaporn","year":"2022","journal-title":"PLOS Neglected Tropical Diseases"},{"issue":"9","key":"pntd.0011596.ref015","doi-asserted-by":"crossref","first-page":"e0239069","DOI":"10.1371\/journal.pone.0239069","article-title":"Combined PCR and MAT improves the early diagnosis of the biphasic illness leptospirosis.","volume":"15","author":"N Philip","year":"2020","journal-title":"PloS one."},{"key":"pntd.0011596.ref016","author":"WW Daniel","year":"2013","journal-title":"Biostatistics: a foundation for analysis in the health sciences"},{"issue":"3","key":"pntd.0011596.ref017","doi-asserted-by":"crossref","first-page":"e0006319","DOI":"10.1371\/journal.pntd.0006319","article-title":"Thai-Lepto-on-admission probability (THAI-LEPTO) score as an early tool for initial diagnosis of leptospirosis: Result from Thai-Lepto AKI study group.","volume":"12","author":"T Sukmark","year":"2018","journal-title":"PLoS Negl Trop Dis."},{"issue":"3","key":"pntd.0011596.ref018","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.diagmicrobio.2009.03.014","article-title":"Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene","volume":"64","author":"RA Stoddard","year":"2009","journal-title":"Diagn Microbiol Infect Dis"},{"key":"pntd.0011596.ref019","year":"2013","journal-title":"Leptospirosis (Leptospira interrogans) 2013 Case Definition"},{"key":"pntd.0011596.ref020","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/978-1-0716-0459-5_1","article-title":"Isolation and Culture of Leptospira from Clinical and Environmental Samples.","volume":"2134","author":"C Goarant","year":"2020","journal-title":"Methods in molecular biology (Clifton, NJ)."},{"issue":"1","key":"pntd.0011596.ref021","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1080\/00365510701666031","article-title":"McNemar \u03c72 test revisited: comparing sensitivity and specificity of diagnostic examinations","volume":"68","author":"A Trajman","year":"2008","journal-title":"Scandinavian Journal of Clinical and Laboratory Investigation"},{"issue":"1","key":"pntd.0011596.ref022","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s40794-021-00154-2","article-title":"Leptospirosis infections among hospital patients, Sarawak, Malaysia.","volume":"7","author":"K-C Hii","year":"2021","journal-title":"Trop Dis Travel Med Vaccines"},{"issue":"1","key":"pntd.0011596.ref023","doi-asserted-by":"crossref","first-page":"5066","DOI":"10.1038\/s41598-022-09079-y","article-title":"Spatial\u2013temporal patterns and risk factors for human leptospirosis in Thailand, 2012\u20132018.","volume":"12","author":"S Chadsuthi","year":"2022","journal-title":"Scientific reports."},{"issue":"1","key":"pntd.0011596.ref024","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/S0021-9975(05)80089-1","article-title":"Kinetics of the local and systemic antibody response to primary and secondary infection with S48 Toxoplasma gondii in sheep","volume":"112","author":"JM Wastling","year":"1995","journal-title":"Journal of comparative pathology"},{"issue":"4","key":"pntd.0011596.ref025","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1038\/s41577-019-0123-x","article-title":"Adaptive immune responses to primary and secondary dengue virus infections.","volume":"19","author":"AL St. John","year":"2019","journal-title":"Nature Reviews Immunology"},{"issue":"2","key":"pntd.0011596.ref026","first-page":"201","article-title":"Development of an immunochromatographic test with anti-LipL32-coupled gold nanoparticles for Leptospira detection.","volume":"37","author":"C Chirathaworn","year":"2014","journal-title":"The new microbiologica."}],"updated-by":[{"DOI":"10.1371\/journal.pntd.0011596","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T00:00:00Z","timestamp":1694476800000}}],"container-title":["PLOS Neglected Tropical Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0011596","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T13:32:08Z","timestamp":1694525528000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pntd.0011596"}},"subtitle":[],"editor":[{"given":"Vasantha Kumari","family":"Neela","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2023,8,25]]},"references-count":26,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2023,8,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pntd.0011596","relation":{},"ISSN":["1935-2735"],"issn-type":[{"value":"1935-2735","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8,25]]}}}