Development and application of a recombinase-aided amplification and lateral flow assay for rapid detection of pseudorabies virus from clinical crude samples
Development and application of a recombinase-aided amplification and lateral flow assay for rapid detection of pseudorabies virus from clinical crude samples
复制标题
重组酶辅助扩增和侧流检测的开发和应用，用于快速检测临床粗样品中的伪狂犬病病毒
DOI：
10.1016/j.ijbiomac.2022.10.153
复制
发表时间：
2022-12-17
影响因子：
8.2
通讯作者：
Zhang，Gaiping
中科院分区：
文献类型：
--
作者：
Guo，Zhenhua;Xing，Guangxu;Zhang，Gaiping
伪狂犬病病毒（Pseudorabiesvirus，PRV）是影响我国养猪业的重要经济病毒。准确、快速、简便的检测是控制和根除PRV的关键。本研究成功地建立了一种检测PRV gE基因的重组酶辅助扩增技术（RAA-LFA）。RAA-LFA与其他猪病原菌均不发生反应，表明该方法具有较好的特异性。该RAA-LFA方法对含有gE基因的标准质粒的检测限（LOD）为21个拷贝/反应。值得注意的是，在无核酸提取条件下，RAA-LFA每次反应可检测低至6.0 × 10050%组织培养感染剂量（TCID 50）的病毒滴度。临床检测结果表明，RAA-LFA检测结果与qPCR检测结果完全一致。总之，开发的PRV RAA-LFA方法为PRV床旁检测（POCT）提供了可接近性、可比准确性和灵敏度工具，这对于设备和人员资源稀缺的地区的PRV控制是有价值的。
Pseudorabies virus (PRV) is an economically important viral agent affecting the swine industry in China. Accurate, rapid and simple detection is critical to PRV control and eradication. In the present study, a visible and low equipment-dependent recombinase-aid amplification assay integrated with lateral flow assay (RAA-LFA) was successfully developed to detect the PRV against the gE gene. The RAA-LFA did not react with the other swine pathogens, indicating the method has a good specificity. The limit of detection (LOD) for this RAA-LFA method was 21 copies per reaction against standard plasmids containing gE gene. Notably, the RAA-LFA can detect as low as 6.0 × 10050 % tissue culture infective dose (TCID50) viral titer per reaction under nucleic-acid-extraction free condition. Clinical detection showed that the results detected by RAA-LFA were completely consistent with that of the qPCR assay. Taken together, the developed PRV RAA-LFA method provides approachability, comparable accuracy and sensitivity tool for PRV point-of-care testing (POCT), which is valuable to PRV control in areas where equipment and personnel resources are scarce.