Establishment and Optimization of Realtime PCR Assays for Detecting Disease in Shrimp Caused by Decapod Iridescent Virus 1

Received: 28-07-2023

Accepted: 26-01-2024

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Issue: Vol. 22 No. 2 (2024)

Section:

CHĂN NUÔI – THÚ Y – THỦY SẢN

How to Cite:

Hoai, T., Huyen, N., Oanh, N., Thang, V., Ngoc, N., Loan, N., … Man, N. (2024). Establishment and Optimization of Realtime PCR Assays for Detecting Disease in Shrimp Caused by Decapod Iridescent Virus 1. Vietnam Journal of Agricultural Sciences, 22(2), 215–225. http://testtapchi.vnua.edu.vn/index.php/vjasvn/article/view/1260

Establishment and Optimization of Realtime PCR Assays for Detecting Disease in Shrimp Caused by Decapod Iridescent Virus 1

Truong Dinh Hoai (*) 1, 2, 3 , Nguyen Thi Huyen , Nguyen Thi Kim Oanh , Vu Dang Thang , Nguyen Dang Hong Ngoc , Nguyen Thanh Loan , Au Xuan Khoa , Vu Thi Lan Huong , Nguyen Thi Thuy Man

  • 1 Khoa Chăn nuôi và NTTS
  • 2 Faculty of Animal Science and Aquaculture, Vietnam National University of Agriculture
  • 3 Khoa Thủy sản, Học viện Nông nghiệp Việt Nam

    • Keywords

    Real-time PCR, procedure establishment, optimization, MCP and ATPase genes, plasmids, DIV1

    Abstract


    The study was conducted to establish, optimize and validate the real-time PCR assay for the detection of Decapod iridescent virus 1(DIV1) and control of this disease in shrimp. The plasmid samples containing two target genes MCP and ATPase of DIV1 were used to establish and optimize the procedure using different probe concentrations 0.1, 0.15 and 0.2µM. The sensitivity, performance, specificity, inter-assay stability, and interlaboratory matching results from nationally and internationally standardized laboratories were used to evaluate the efficacy and validity of these real-time PCR assays. The results of real-time PCR using plasmids containing target MCP and ATPase genes at a probe concentration of 0.2µM revealed the best detection ability. The reaction detection limits were 13.6 and 14.3 plasmid copies/reaction with the efficiency at 98.9% and 92.6%, respectively. The assays exhibited high specificity (no reaction to all shrimp-positive samples for WSSV, IHHNV, AHPND, and EHP) and high stability (CV < 15%). The study results showed that two real-time PCR assays using plasmids carrying target MCP and ATPase genes of DIV1 established in this study could ensure reliability and applicability to laboratories for screening and controlling DIV1 disease in Vietnam.

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