福建农业学报2025，Vol.40Issue(3)：245-252,8.DOI:10.19303/j.issn.1008-0384.2025.03.004
基于RT-RPA-微流控芯片的牛副流感病毒3型可视化快速检测方法
RT-RPA Microfluidic Chips-based Visualizable Rapid Method for Detecting Bovine Parainfluenza Virus Type 3
摘要
Abstract
[Objective]A rapid and visualizable method for detecting bovine parainfluenza virus type 3(BPIV3)was developed utilizing the reverse transcription-recombinase polymerase amplification(RT-RPA),magnetic capture,and microfluidic chips.[Methods]Specific primers and probes were designed based on the BPIV3 gp3 gene.The bovine viral diarrhea virus(BVDV),bovine adenovirus type 3(BADV3),and bovine infectious rhinotracheitis virus(IBRV)were used as control to determine assay specificity.Assay sensitivity was examined on a 10×dilution of RNA extraction solution of the standard BPIV3 strain.Fifty-six serum and nasal swab samples from the cattle suspected of acute BPIV3 infection were collected for detection using the newly established method.Test results were compared with those obtained on nasal swab samples by RT-PCR.[Results]The amplicon from the RNA extract of the standard BPIV3 strain sized 404 bp,with none detected in control,obtained by the assay.A 100%homology with the highly conserved gp3 gene NC_002161.1 of BPIV3 was shown.At different concentrations of the extract,the lowest assay detection limit was 2.26×103 copies·µL-1.From the 56 serum samples,the cows,coded SD0078,SD0114,SD0319,SD0601,SD0714,and SD0755,were identified to be positively infected by BPIV3.The results agreed with what were obtained from RT-PCR on the nasal swab isolation samples.It took the assay merely 92 min from sample preparation to data acquisition.[Conclusion]A rapid and visualizable method for BPIV3 detection was developed using RT-RPA,magnetic capture,and microfluidic chips.The assay was high on specificity,sensitivity,and applicability for the intended purpose.关键词
牛副流感病毒3/逆转录重组酶聚合酶扩增技术/磁捕获/微流控芯片技术Key words
bovine respiratory virus 3/reverse transcription-recombinase polymerase amplification/magnetic capture/microfluidic chips分类
畜牧业引用本文复制引用
张铁男,侯梦媛,毛晓庆..基于RT-RPA-微流控芯片的牛副流感病毒3型可视化快速检测方法[J].福建农业学报,2025,40(3):245-252,8.基金项目
哈尔滨商业大学企业技术开发项目(HSDHX202104061) （HSDHX202104061）
