Establishment and application of a real-time fluorescence quantitative PCR assay for detecting EAPV in Passiflora edulis
4 affiliations
- 1Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences/Yunnan Provincial Key Laboratory of Agricultural Biotechnology, Kunming, China
- 2Forestry College/Key Laboratory of Forest Disaster Warning and Control in Yunnan Province, Faculty of Biodiversity Conservation, Southwest Forestry University, Kunming, China
- 3School of Ethnic Medicine, Yunnan Minzu University, Kunming, China
- 4Riphah International University, Faisalabad Campus, Faisalabad, Pakistan
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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BibTeX
@article{zhou2026establishment,
title = {{Establishment and application of a real-time fluorescence quantitative PCR assay for detecting EAPV in Passiflora edulis}},
author = {Hanye Zhou and Jie Zhang and Yu Li and Ziran Gao and Muhammad Qasim Aslam and Jin Xu and Yongdui Chen},
journal = {PLOS One},
year = {2026},
month = sep,
doi = {10.1371/journal.pone.0356266},
url = {https://doi.org/10.1371/journal.pone.0356266},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Establishment and application of a real-time fluorescence quantitative PCR assay for detecting EAPV in Passiflora edulis
AU - Hanye Zhou
AU - Jie Zhang
AU - Yu Li
AU - Ziran Gao
AU - Muhammad Qasim Aslam
AU - Jin Xu
AU - Yongdui Chen
PY - 2026
DA - 2026/09/08
JO - PLOS One
DO - 10.1371/journal.pone.0356266
UR - https://doi.org/10.1371/journal.pone.0356266
AB - 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.
SN - 1932-6203
ER -