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A TaqMan probe-based RT-qPCR for the detection of brine shrimp reovirus 1

文献类型： 外文期刊

作者： Li, Xuan 1 ; Wang, Guohao 2 ; Lou, Haoyu 2 ; Zhou, Chengyan 2 ; Meng, Xiangmin 1 ; Wang, Xinping 1 ; Dong, Xuan 1 ;

作者机构： 1.Qingdao Univ Sci & Technol, Coll Biol Engn, Qingdao 266042, Peoples R China

2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Qingdao Marine Sci & Technol Ctr,Lab Marine Fisher, Qingdao 266071, Peoples R China

3.Chinese Acad Fishery Sci, Minist Agr & Rural Affairs, Key Lab Maricultural Organism Dis Control, Qingdao 266071, Peoples R China

4.Chinese Acad Fishery Sci, Qingdao Key Lab Mariculture Epidemiol & Biosecur, Qingdao 266071, Peoples R China

5.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai 201306, Peoples R China

6.Huzhou Univ, Coll Life Sci, Huzhou 313000, Peoples R China

关键词： Brine shrimp reovirus 1; Detection; Real-time RT-qPCR; Artemia

期刊名称：AQUACULTURE （ 影响因子：3.9； 五年影响因子：4.4 ）

ISSN： 0044-8486

年卷期： 2025 年 604 卷

页码：

收录情况： SCI

摘要： Brine shrimp (Artemia) are widely utilized as a vital live feed in the aquaculture industry. Recently, we found a novel reovirus- brine shrimp reovirus 1 (BSR1) in brine shrimp. Considering the potential impact of reovirus on the industry, it is necessary to develop a highly sensitive and specific detection method in advance for identifying BSR1. In this study, we established a real-time quantitative reverse transcription PCR (RT-qPCR) method based on TaqMan probe. The detection limit of this method is as low as 4.4 x 101 copies/reaction. The standard curve between 4.4 x 101 to 4.4 x 109 copies/reaction showed a high correlation coefficient (R2 = 0.997). This newly developed method was used to screen for BSR1 in brine shrimp cyst samples from Africa, Asia, Europe, Oceania, South America and North America, and the results showed that BSR1 was detected in samples from Asia and North America. The brine shrimp cyst samples tested covered seven different species, with BSR1-positive samples found in A. franciscana, A. urmiana, A. sinica, and parthenogenetic Artemia lineage. The diagnostic sensitivity (DSe) and diagnostic specificity (DSp) values of the newly developed RT-qPCR method, when compared to the high-throughput sequencing, were determined to be 100 % and 88 %. This newly developed RT-qPCR method can serve as a crucial tool for detecting BSR1, facilitating early warning and swift response to viruses carried by brine shrimp, thereby effectively mitigating the risk of pathogen transmission to cultured animals.

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