{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T13:24:34Z","timestamp":1790083474153,"version":"4.0.1"},"reference-count":47,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,2,14]],"date-time":"2025-02-14T00:00:00Z","timestamp":1739491200000},"content-version":"vor","delay-in-days":13,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2025,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:label\/>\n                    <jats:p>\n                      Globally \u2264\u20094 billion of the population are at potential risk of contracting dengue virus (DENV) infection. Seasonal outbreaks of dengue are frequently reported causing a high healthcare burden. Undiagnosed DENV can lead to severe morbidity and mortality. Early diagnosis of DENV relies on molecular methods, which are impractical in resource\u2010constrained settings (RCSs). Dengue can be caused by any of the four distinct DENV serotypes. Therefore, a simple method for rapid diagnosis of Pan\u2010DENV serotypes is of utmost importance at RCSs. A fluorescence detection platform for Pan\u2010DENV using RT\u2010RPA and CRISPR\/Cas12a was developed targeting nonstructural 1 (\n                      <jats:italic>NS1<\/jats:italic>\n                      ) gene for DENV\u20101, 2, and 3, and envelope (\n                      <jats:italic>E<\/jats:italic>\n                      ) gene for DENV\u20102. Further, crRNA specific to DENV serotypes were designed to facilitate CRISPR\/Cas12a detection. Analytical sensitivity was determined using synthetic RNA and DENV serotypes genome. Clinical validation of the assay was performed using RNA extracted from AES\/AFI clinical samples. The developed CRISPR\/Cas12a\u2010based detection platform can detect all four serotypes of DENV viz 1\u22124 in a single pot using fluorescence detection. This assay showed the limit of detection \u2265\u2009781\u2009zg reaction\n                      <jats:sup>\u2212<\/jats:sup>\n                      <jats:sup>1<\/jats:sup>\n                      , \u2265\u20091.81\u2009ag reaction\n                      <jats:sup>\u22121<\/jats:sup>\n                      , \u2265\u200962.5\u2009fg reaction\n                      <jats:sup>\u22121<\/jats:sup>\n                      , and \u2265\u20092.5\u2009pg reaction\n                      <jats:sup>\u22121<\/jats:sup>\n                      for synthetic DENV\u20101, DENV\u20102, DENV\u20103, and DENV\u20104 template, respectively. Our assay demonstrated the analytic sensitivity of \u2265\u200910\u2009ng reaction\n                      <jats:sup>\u22121<\/jats:sup>\n                      for DENV\u20101 and DENV\u20104, and \u2265\u20090.5\u2009ng\u2009reaction\n                      <jats:sup>\u22121<\/jats:sup>\n                      for DENV\u20103 and DENV\u20104 genomes. This assay showed no cross\u2010reactivity with other related etiologies tested causing AFI\/AES. With 76 clinical samples (DENV PCR positive\u2009=\u200916, DENV PCR negative\u2009=\u200960), the assay demonstrated 93.7% sensitivity and 100% specificity with an overall accuracy of 98.7% for detection of the Pan\u2010DENV serotypes. Our assay displayed comparable results to that of RT\u2010PCR. The ease of interpretation and rapid detection of the Pan\u2010DENV, represents the potential of the developed assay as an ideal point\u2010of\u2010care test. This assay upon field\u2010deployment could help in reducing healthcare burden, provide differential diagnosis and support initiating early and prompt treatment to patients at RCS.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/jmv.70219","type":"journal-article","created":{"date-parts":[[2025,2,14]],"date-time":"2025-02-14T03:19:37Z","timestamp":1739503177000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["RT\u2010RPA Assisted CRISPR\/Cas12a Based One\u2010Pot Rapid and Visual Detection of the Pan\u2010Dengue Virus"],"prefix":"10.1002","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7497-0291","authenticated-orcid":false,"given":"Pooja","family":"Bhardwaj","sequence":"first","affiliation":[{"name":"JE\u2010AES Apex Laboratory, ICMR\u2010Regional Medical Research Centre BRD Medical College Campus Gorakhpur 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