{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:52:34Z","timestamp":1783039954617,"version":"3.54.6"},"update-to":[{"DOI":"10.1371\/journal.pntd.0012024","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2024,7,26]],"date-time":"2024-07-26T00:00:00Z","timestamp":1721952000000}}],"reference-count":39,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T00:00:00Z","timestamp":1721088000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key R & D Projects of Hubei Province","award":["2022BCA019"],"award-info":[{"award-number":["2022BCA019"]}]},{"name":"Hubei Provincial Public Leadership Talents Project","award":["SGGLJ202201"],"award-info":[{"award-number":["SGGLJ202201"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:p>Central China has been reported to be one of the most important endemic areas of zoonotic infection by spotted fever group rickettsiae (SFGR), severe fever with thrombocytopenia syndrome virus (SFTSV) and hantaan virus (HTNV). Due to similar clinical symptoms, it is challenging to make a definite diagnosis rapidly and accurately in the absence of microbiological tests. In the present study, an all-in-one real-time PCR assay was developed for the simultaneous detection of nucleic acids from SFGR, SFTSV and HTNV. Three linear standard curves for determining SFGR-<jats:italic>ompA<\/jats:italic>, SFTSV-L and HTNV-L were obtained within the range of 10<jats:sup>1<\/jats:sup>\u221210<jats:sup>6<\/jats:sup> copies\/\u03bcL, with the PCR amplification efficiencies ranging from 93.46% to 96.88% and the regression coefficients R<jats:sup>2<\/jats:sup> of &gt;0.99. The detection limit was 1.108 copies\/\u03bcL for SFGR-<jats:italic>ompA<\/jats:italic>, 1.075 copies\/\u03bcL for SFTSV-L and 1.006 copies\/\u03bcL for HTNV-L, respectively. Both the within-run and within-laboratory coefficients of variation on the cycle threshold (Ct) values were within the range of 0.53%-2.15%. It was also found there was no statistical difference in the Ct values between single template and multiple templates (P<jats:sub><jats:bold>SFGR-<jats:italic>ompA<\/jats:italic><\/jats:bold><\/jats:sub> = 0.186, P<jats:sub>SFTSV-L<\/jats:sub> = 0.612, P<jats:sub>HTNV-L<\/jats:sub> = 0.298). The sensitivity, specificity, positive and negative predictive value were all 100% for determining SFGR-<jats:italic>ompA<\/jats:italic> and SFTSV-L, 97%, 100%, 100% and 99.6% for HTNV-L, respectively. Therefore, the all-in-one real-time PCR assay appears to be a reliable, sensitive, rapid, high-throughput and low cost-effective method to diagnose the zoonotic infection by SFGR, SFTSV and HTNV.<\/jats:p>","DOI":"10.1371\/journal.pntd.0012024","type":"journal-article","created":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T17:41:58Z","timestamp":1721151718000},"page":"e0012024","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Development of an all-in-one real-time PCR assay for simultaneous detection of spotted fever group rickettsiae, severe fever with thrombocytopenia syndrome virus and hantaan virus prevalent in central 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