Xu Ting, Tan Rongxiang, Zhu Yutao, Ye Jian
School of Life and Environmental Sciences, Shaoxing University, Shaoxing 312000, China.
School of Life and Environmental Sciences, Shaoxing University, Shaoxing 312000, China.
J Invertebr Pathol. 2023 Nov;201:107998. doi: 10.1016/j.jip.2023.107998. Epub 2023 Oct 5.
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 (R = 1.000) and good repeatability in detecting standards of DIV1 MCP fragments ranging from 6.2 × 10 to 6.2 × 10 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.
十足目虹彩病毒1（DIV1）是一种新兴病原体，主要威胁十足目甲壳类动物，可导致高死亡率并造成巨大经济损失。在本研究中，针对DIV1的主要衣壳蛋白（MCP）基因设计了一对特异性引物，并建立了基于SYBR Green I的实时荧光定量PCR方法。该方法在检测浓度范围为6.2×10至6.2×10个DNA拷贝/微升的DIV1 MCP片段标准品时，显示出良好的线性关系（R = 1.000）和重复性。特异性分析表明，该实时荧光定量PCR对DIV1具有特异性，不与其他常见虾类病原体或健康虾的DNA发生反应。敏感性分析表明，该实时荧光定量PCR能够在35个循环内有效检测低至62拷贝/微升的DIV1 DNA。总之，所建立的实时荧光定量PCR为DIV1提供了一种高效、灵敏且可靠的检测方法。
