Establishment of a SYBR Green I-based real-time PCR for the detection of decapod iridescent virus 1

Ting Xu; Rongxiang Tan; Yutao Zhu; Jian Ye

doi:10.1016/j.jip.2023.107998

Establishment of a SYBR Green I-based real-time PCR for the detection of decapod iridescent virus 1

J Invertebr Pathol. 2023 Nov:201:107998. doi: 10.1016/j.jip.2023.107998. Epub 2023 Oct 5.

Authors

Ting Xu¹, Rongxiang Tan², Yutao Zhu², Jian Ye³

Affiliations

¹ School of Life and Environmental Sciences, Shaoxing University, Shaoxing 312000, China. Electronic address: xuting@usx.edu.cn.
² School of Life and Environmental Sciences, Shaoxing University, Shaoxing 312000, China.
³ Hangzhou Centre for Agricultural Technology Extension, Hangzhou 310017, China. Electronic address: yejian1015@163.com.

PMID: 37802421
DOI: 10.1016/j.jip.2023.107998

Abstract

Decapod iridescent virus 1 (DIV1) is an emerging pathogen that mainly threatens decapod crustaceans, causing high mortalities and leading to huge economic losses. In this study, a pair of specific primers were designed for the major capsid protein (MCP) gene of DIV1, and a SYBR Green I-based real-time PCR method was developed. The method displayed good linearity (R² = 1.000) and good repeatability in detecting standards of DIV1 MCP fragments ranging from 6.2 × 10¹ to 6.2 × 10⁸ DNA copies/μl. Specificity analysis revealed that the real-time PCR was specific for DIV1 and did not react with other common shrimp pathogens or healthy shrimp DNA. Sensitivity analysis revealed that the real-time PCR could efficiently detect DIV1 DNA as low as 62 copies/μl within 35 cycles. In summary, the established real-time PCR provides an efficient, sensitive, and reliable detection method for DIV1.

Keywords: Decapod iridescent virus 1; Litopenaeus vannamei; Real-time PCR.

MeSH terms

Animals
Benzothiazoles*
DNA
Decapoda*
Real-Time Polymerase Chain Reaction / methods
Sensitivity and Specificity

Substances

SYBR Green I
Benzothiazoles
DNA