Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

牛病毒性腹泻病毒1型RT-RAA快速诊断方法的建立_范颖

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To address the challenge of rapid on-site diagnosis of Bovine Viral Diarrhea Virus (BVDV) and enhance emergency response efficiency, specific primers and probes were designed based on the conserved 5'-UTR region of the BVDV type 1 genome. This led to the development of a recombinase-aided isothermal nucleic acid amplification method for BVDV type 1 (reverse transcription recombinase aid amplification, RT-RAA). The specificity, sensitivity, and reproducibility of this method were subsequently evaluated. The results demonstrated that the assay can detect BVDV type 1 with a detection limit as low as 1.2 × 10² copies/μL within a 20-minute incubation period at a constant temperature of 39°C. Moreover, intra-assay repeatability tests conducted across three concentration gradients yielded coefficient of variation (CV) values of 3.24%, 9.48%, and 8.79%, respectively— all remaining below the 10.00% threshold, indicating excellent reproducibility. Importantly, the developed method showed no cross-reactivity with other pathogens, including Classical Swine Fever Virus, Bovine Coronavirus, and Bovine Infectious Rhinotracheitis Virus. When applied to 66 clinical samples, the RT-RAA method exhibited a high concordance rate of 98.48% compared to fluorescence quantitative PCR results, with a Kappa value of 0.881 (P < 0.001). These findings underscore that the RT-RAA assay established in this study is highly specific, sensitive, and easy to perform, making it an ideal tool for rapid on-site detection of BVDV type 1.

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非洲猪瘟病毒实时荧光RAA检测方法的建立-南京农大&动卫

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重组酶介导的油菜茎基溃疡病...荧光核酸扩增检测体系的建立_陈吴健