Center of Inspection and Quarantine, Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang 050051, China; Hebei Academy of Science and Technology for Inspection and Quarantine, Shijiazhuang 050051, China.
College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China.
Mol Cell Probes. 2018 Jun;39:41-46. doi: 10.1016/j.mcp.2018.04.004. Epub 2018 Apr 27.
A visible and equipment-free recombinase polymerase amplification assay combined with a lateral flow strip (LFS RPA) was developed to detect canine parvovirus type 2 (CPV-2), which is the etiological agent of canine parvovirus disease. The CPV-2 LFS RPA assay was developed based on the VP2 gene and is performed in a closed fist using body heat for 15 min; the products are visible to the naked eye on the LFS within 5 min. The assay could detect CPV-2a, CPV-2b and CPV-2c, and there was no cross-reaction with the other viruses tested. Using the standard CPV-2 DNA as a template, the analytical sensitivity was 1.0 × 10 copies per reaction, which was the same result as that of a real-time PCR. The assay performance was further evaluated by testing 60 canine fecal samples, and CPV-2 DNA was detected in 46 samples (76.7%, 46/60) by LFS RPA, which was the same result as that of the real-time PCR assay and higher than that of the SNAP method (48.3%, 29/60). The novel CPV-2 LFS RPA assay is an attractive and promising tool for rapid and convenient diagnosis of CPV disease, especially cage side and in underequipped laboratories.
一种可见且无需设备的重组酶聚合酶扩增检测方法（LFS RPA）与侧向流条带（LFS RPA）结合，用于检测犬细小病毒 2 型（CPV-2），CPV-2 是犬细小病毒病的病原体。CPV-2 LFS RPA 检测方法基于 VP2 基因建立，在封闭的拳手中使用体热进行 15 分钟反应；产物在 5 分钟内可在 LFS 上肉眼可见。该检测方法可检测 CPV-2a、CPV-2b 和 CPV-2c，与测试的其他病毒无交叉反应。使用标准 CPV-2 DNA 作为模板，分析灵敏度为 1.0×10 拷贝/反应，与实时 PCR 结果相同。通过检测 60 份犬粪便样本进一步评估了该检测方法的性能，LFS RPA 检测到 46 份样本（76.7%，46/60）中存在 CPV-2 DNA，与实时 PCR 检测结果相同，且高于 SNAP 方法（48.3%，29/60）。新型 CPV-2 LFS RPA 检测方法是一种快速、方便诊断 CPV 病的有吸引力和有前途的工具，尤其是在笼边和设备不足的实验室。
