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Development and application of reverse transcriptase droplet digital PCR technology for sensitive detection of BVDV-1 and BTV in bovine semen

文献类型： 外文期刊

作者： Yu, Zhichao 1 ; Chen, Linjun 2 ; Cui, Qiang 2 ; Yan, Han 2 ; Li, Junyan 3 ; Luo, Xiaoping 3 ; Li, Yingying 4 ; Ju, Xianghong 1 ; Yong, Yanhong 1 ; Zhao, Namula 1 ; Zhao, Zhiguo 2 ;

作者机构： 1.Guangdong Ocean Univ, Coll Coastal Agr Sci, Dept Vet Med, Zhanjiang 524088, Guangdong, Peoples R China

2.Technol Ctr, Hohhot 010020, Inner Mongolia, Peoples R China

3.Inner Mongolia Acad Agr & Anim Husb Sci, Vet Res Inst, Key Lab Grass Feeding Livestock Healthy Breeding &, Hohhot 010031, Inner Mongolia, Peoples R China

4.Anim & Plant Quarantine & Anim Dis Prevent & Contr, Baotou 014016, Inner Mongolia, Peoples R China

关键词： Bovine viral diarrhoea virus genotype 1; Bluetongue virus; Droplet digital PCR; Bovine semen

期刊名称：BMC VETERINARY RESEARCH （ 影响因子：2.6； 五年影响因子：2.7 ）

ISSN：

年卷期： 2025 年 21 卷 1 期

页码：

收录情况： SCI

摘要： BackgroundBovine viral diarrhoea virus genotype 1 (BVDV-1) and bluetongue virus (BTV) are potent viral pathogens that may be transmitted through semen, resulting in the spread of diseases via artificial insemination. Thus, establishing an early detection method for BVDV-1 and BTV infection is important for the trading of semen. In this study, we developed two RT-ddPCR methods to detect BVDV-1 and BTV, and each method was evaluated for repeatability, limit of detection and specificity. The sensitivity of these methods was compared with that of RT-qPCR (WOAH) by analysing clinical samples.ResultsThe RT-ddPCR results revealed that both methods exhibited good repeatability at low concentrations, with detection limits of 1.05 copies/mu L and 0.662 copies/mu L per reaction for BVDV-1 and BTV, respectively; additionally, both methods exhibited high specificity and did not exhibit cross-reaction with other important semen-transmitted pathogens. Eighty bovine semen samples and twenty mixed semen samples were tested. The results revealed that the positivity rates of BVDV-1 and BTV RT-ddPCR (25% and 23%, respectively) were greater than those of RT-qPCR (19% and 18%, respectively).ConclusionsRT-ddPCR was highly sensitive for detecting low concentrations of BVDV-1 and BTV in clinical samples and could be a good supplement for qPCR testing.

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