基于侧向流纸条的重组酶聚合酶扩增检测试剂盒的研制及其在山羊粪便中检测捻转血矛线虫的应用。
Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces.
机构信息
Laboratory of Guangdong Province and Hong Kong Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou, 510642, China.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China.
出版信息
Korean J Parasitol. 2021 Apr;59(2):167-171. doi: 10.3347/kjp.2021.59.2.167. Epub 2021 Apr 22.
Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.
捻转血矛线虫病仍然是小反刍动物的一个重大问题。在这项研究中，建立了重组酶聚合扩增（RPA）联合侧流条带（LFS-RPA）检测试剂盒，用于快速检测山羊粪便中的捻转血矛线虫。该检测试剂盒使用针对 ITS-2 基因中特定序列的引物和探针。我们比较了 LFS-RPA 检测试剂盒与 PCR 检测试剂盒的性能。LFS-RPA 的检测限为 10 fg DNA，比 PCR 获得的最低检测限低 10 倍。在 24 份山羊粪便样本中，LFS-RPA 检测试剂盒检测到 H. contortus DNA 的灵敏度为 95.8%，而 PCR 检测试剂盒的灵敏度为 79.1%。LFS-RPA 检测试剂盒未检测到其他相关寄生虫的 DNA，并且对山羊粪便中存在的抑制剂具有足够的耐受性。总之，我们的结果表明，LFS-RPA 检测试剂盒具有较高的诊断准确性，可用于快速检测 H. contortus，值得进一步评估。
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