A lateral flow dipstick combined with reverse transcription recombinase polymerase amplification for rapid and visual detection of the BVDV and BPIV3

文献类型： 外文期刊

第一作者： Yang, Sen

作者： Yang, Sen;Wang, Qian-Ying;Tan, Bin;Shi, Peng-Fei;Qiao, Lian-Jiang;Li, Zhi-Jie;Liu, Ke-Xin;Cao, Zhi-Gang;Zhang, Shu-Qin;Sun, Fei-Yan

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关键词： Bovine viral diarrhea virus; Bovine parainfluenza virus type 3; Reverse transcription recombinase polymerase amplification; Lateral flow dipstick

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.623； 五年影响因子：2.239 ）

ISSN： 0166-0934

年卷期： 2022 年 299 卷

页码：

收录情况： SCI

摘要： Bovine respiratory disease complex (BRDC) is a serious disease affecting feedlot cattle in China and likely other places worldwide. Bovine viral diarrhea virus (BVDV) and bovine parainfluenza virus type 3 (BPIV3) are principally responsible for causing BRDC, and are a major strain to the industrial economy. Eradication of these viruses/disease requires swift viral identification and treatment. Hence, this study established a fast and easy procedure of BVDV and BPIV3 identification that employs reverse transcription recombinase polymerase amplification (RT-RPA) and lateral flow dipstick (LFD), and uses primers and lateral flow (LF) probe targeting the 5 '-UTR gene of BVDV and phosphoprotein P gene of BPIV3, respectively. Our assay was able to successfully amplify BVDV and BPIV3 RNA within 25 min at 35 degrees C using RT-RPA, with products visible on the LFD within 5 min at room temperature (RT). The lowest detection limits were 50 RNA molecules for BVDV and 34 RNA molecules for BPIV3 per reaction. We also demonstrated that the established dual RT-RPA LFD assay was precise and targeted, harboring excellent potential to become an onsite molecular diagnostic tool in the detection of BVDV and BPIV3. This method can detect BVDV (Pestivirus A, B) and BPIV3, and exhibit no cross-reaction with other viruses like the classical swine fever virus (CSFV) and infectious bovine rhinotracheitis virus (IBRV). The assay performance was further assessed with clinical samples, and demonstrated good performance in comparison to real-time RT-PCR (RT-qPCR). Moreover, the RT-RPA LFD assay was comparitively rapid and required minimal training.

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