ArticlePloS one2026
Establishment and application of a real-time fluorescence quantitative PCR assay for detecting EAPV in Passiflora edulis.
Article in PloS one, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
East Asian Passiflora virus (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 μmol/L and an annealing temperature of 60°C. The primers exhibited high specificity, generating a standard curve with an amplification efficiency of 90.9% and a coefficient of determination (R2) of 0.992. The limit of detection was 11.41 × 102 copies/μL, 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.
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