{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T18:08:19Z","timestamp":1788890899046,"version":"build-2803163510"},"reference-count":28,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Industrial Innovation Talent Project of \u201cYunnan Revitalization Talent Support Project\u201d","award":["XDYC-CYCX-2022-0065"],"award-info":[{"award-number":["XDYC-CYCX-2022-0065"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31860498"],"award-info":[{"award-number":["31860498"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    <jats:italic>East Asian Passiflora virus<\/jats:italic>\n                    (EAPV) is a significant viral pathogen prevalent across passionfruit cultivation regions in China and causes substantial economic losses to the passionfruit industry. The development of a sensitive, rapid, and accurate diagnostic method is essential for virus identification and epidemiological surveillance to support effective disease management strategies. In this study, a SYBR Green-based real-time quantitative PCR (qPCR) assay was developed for EAPV detection using a specific primer set targeting the viral coat protein (CP) gene. The assay was optimized with a primer concentration of 0.2 \u03bcmol\/L and an annealing temperature of 60\u00b0C. The primers exhibited high specificity, generating a standard curve with an amplification efficiency of 90.9% and a coefficient of determination (R\n                    <jats:sup>2<\/jats:sup>\n                    ) of 0.992. The limit of detection was 11.41\u2009\u00d7\u200910\n                    <jats:sup>2<\/jats:sup>\n                    copies\/\u03bcL, representing a 1000-fold greater sensitivity than conventional PCR. Moreover, viral accumulation was successfully detected in both inoculated and systemic leaves of passionfruit plants. In 2025, a total of 120 suspected virus-infected passionfruit samples were collected from Yunnan Province, China, and all samples tested positive for EAPV using the developed qPCR assay. Overall, the SYBR Green-based qPCR method established in this study demonstrated high specificity and sensitivity, providing a reliable tool for rapid EAPV diagnosis and epidemiological investigations.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0356266","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T17:32:47Z","timestamp":1788888767000},"page":"e0356266","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment and application of a real-time fluorescence quantitative PCR assay for detecting EAPV in Passiflora edulis"],"prefix":"10.1371","volume":"21","author":[{"given":"Hanye","family":"Zhou","sequence":"first","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]},{"name":"Forestry College\/Key Laboratory of Forest Disaster Warning and Control in Yunnan Province, Faculty of Biodiversity Conservation, Southwest Forestry University","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Zhang","sequence":"additional","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Li","sequence":"additional","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziran","family":"Gao","sequence":"additional","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]},{"name":"School of Ethnic Medicine, Yunnan Minzu University","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Qasim","family":"Aslam","sequence":"additional","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]},{"name":"Riphah International University, Faisalabad Campus","place":["Faisalabad, Pakistan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Xu","sequence":"additional","affiliation":[{"name":"Forestry College\/Key Laboratory of Forest Disaster Warning and Control in Yunnan Province, Faculty of Biodiversity Conservation, Southwest Forestry University","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-7139-3353","authenticated-orcid":true,"given":"Yongdui","family":"Chen","sequence":"additional","affiliation":[{"name":"Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences\/Yunnan Provincial Key Laboratory of Agricultural Biotechnology","place":["Kunming, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"pone.0356266.ref001","doi-asserted-by":"crossref","first-page":"617","DOI":"10.3389\/fphar.2020.00617","article-title":"Passiflora edulis: An Insight Into Current Researches on Phytochemistry and Pharmacology","volume":"11","author":"X He","year":"2020","journal-title":"Front Pharmacol"},{"issue":"8","key":"pone.0356266.ref002","doi-asserted-by":"crossref","first-page":"2307","DOI":"10.1094\/PDIS-09-22-2167-SC","article-title":"Occurrence and Distribution of Major Viruses Infecting Passion Fruit in Guizhou Province, China, and Molecular Characterization of Two Potyviruses","volume":"107","author":"X Fu","year":"2023","journal-title":"Plant Dis"},{"issue":"10","key":"pone.0356266.ref003","first-page":"839","article-title":"Research Progress on Viruses of Passiflora edulis","volume":"13","author":"W Wu","year":"2024","journal-title":"Biology 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Mohamed","family":"Amer","sequence":"first","affiliation":[{"name":"Cairo University Faculty of Veterinary Medicine","place":["EGYPT"]}],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":28,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9,8]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0356266","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]}}}