中国动物传染病学报2025，Vol.33Issue(2)：80-88,9.
基于RTCPA-CRISPR/Cas12a的牛病毒性腹泻病毒检测方法的建立及应用
Development and Application of RT-CPA-CRISPR/Cas12a Method for Detection of BVDV
摘要
Abstract
To develop an RT-CPA-CRISPR/Cas12a detection method for Bovine viral diarrhea virus(BVDV),this study designed CPA-specific primers based on the E0 protein gene sequence of BVDV.Additionally,crRNA targeting RT-RAA amplification products was designed following CRISPR/Cas12a principles.The RT-CPA-CRISPR/Cas12a reaction system was optimized,and clinical samples were tested using the established method.Results demonstrated that the detection limit of this method for BVDV was 8.0 copies/μL,with high specificity.Testing of 42 clinical samples revealed high consistency(92.86％ concordance rate)with conventional PCR methods.This study successfully established an RT-CPA-CRISPR/Cas12a detection method for BVDV,providing a technical tool for the diagnosis of this disease.关键词
牛病毒性腹泻病毒/交叉引物恒温扩增技术/CRISPR/Cas12a/灵敏度/特异性Key words
Bovine viral diarrhea virus/cross-priming amplification/CRISPR/Cas12a/sensitivity/specificity分类
农业科技引用本文复制引用
蒙琦,常亮,杨明,曹映辉,贺泂杰..基于RTCPA-CRISPR/Cas12a的牛病毒性腹泻病毒检测方法的建立及应用[J].中国动物传染病学报,2025,33(2):80-88,9.基金项目
平凉市重点研发计划项目(PL-STK-2023A-037) （PL-STK-2023A-037）
平凉市重点研发计划项目(PL-STK-2024A-028) （PL-STK-2024A-028）
甘肃省农业农村厅科技计划项目(GNKJ-2022-39) （GNKJ-2022-39）
甘肃省科技计划项目-重点研发计划(21YF5NA143) （21YF5NA143）
