{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T01:14:23Z","timestamp":1792631663516,"version":"4.5.0"},"reference-count":32,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"3","content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2022,7]]},"abstract":"<jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Background &amp; objectives:<\/jats:italic>\n                      Recently, the incidences of chikungunya, dengue and Zika infections have increased due to globalization and urbanization. It is vital that reliable detection tools become available to assess the viral prevalence within mosquito populations.\n                    <\/jats:p>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Methods:<\/jats:italic>\n                      Based on the previous publications on clinical diagnosis in human infections, for the first time, we described a customized triplex RT-qPCR protocol for simultaneous detection of chikungunya virus (CHIKV), dengue virus serotypes 1-4 (DENV1-4) and Zika virus (ZIKV) in mosquitoes.\n                    <\/jats:p>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Results:<\/jats:italic>\n                      In preliminary assessment to determine the specificity and sensitivity of primers and probes, all six targets were detected individually with the following thresholds as indicated by calculated pfu equivalents: 3.96x10\n                      <jats:sup>0<\/jats:sup>\n                      for CHIKV, 3.80x10\n                      <jats:sup>1<\/jats:sup>\n                      for DENV1, 3.20x10\n                      <jats:sup>1<\/jats:sup>\n                      for DENV2, 8.00x10\n                      <jats:sup>4<\/jats:sup>\n                      for DENV3, 1.58x10\n                      <jats:sup>0<\/jats:sup>\n                      for DENV4, and 6.20x10\n                      <jats:sup>0<\/jats:sup>\n                      for ZIKV When tested in a full combination of six targets (CDZ mix), CHIKV, DENV1-4 mix or ZIKV were all detected with the thresholds of 1.32x10\n                      <jats:sup>0<\/jats:sup>\n                      for CHIKV, 3.79x10\n                      <jats:sup>0<\/jats:sup>\n                      for DENV1-4 and 2.06x10\n                      <jats:sup>0<\/jats:sup>\n                      for ZIKV All targets, individually or in full combination were detected in the mixtures of\n                      <jats:italic toggle=\"yes\">Aedes aegypti<\/jats:italic>\n                      (L.) homogenate and viral lysates. A robust evaluation with three replicates in each of three plates for CHIKV, DENV1-4 and ZIKV individually or in full combination was conducted. In individual assays, CHIKV was detected to 3.96x10\n                      <jats:sup>-1<\/jats:sup>\n                      , DENV1-4 to 1.14x10\n                      <jats:sup>0<\/jats:sup>\n                      and ZIKV to 3.20x10\n                      <jats:sup>0<\/jats:sup>\n                      . In full combination assays, CHIKV was detected to 1.32x10\n                      <jats:sup>4<\/jats:sup>\n                      , DENV1-4 to 3.79x10\n                      <jats:sup>1<\/jats:sup>\n                      and ZIKV to 1.07x10\n                      <jats:sup>0<\/jats:sup>\n                      .\n                    <\/jats:p>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Interpretation &amp; conclusion:<\/jats:italic>\n                      This triplex RT-qPCR assay appears to consistently detect all six targets and does not cross react with\n                      <jats:italic toggle=\"yes\">Ae. aegypti<\/jats:italic>\n                      homogenate, making it a feasible, practical, and immediately adoptable protocol for use among vector control and other entities, particularly in the endemic areas of CHIKV, DENVs and ZIKV.\n                    <\/jats:p>\n                  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