Abstract: Objective　To develop a rapid test for detection of Schistosoma japonicum specific gene fragments based on the recombinase⁃aided isothermal amplification assay (RAA) and nucleic acid dipstick test. Methods　The S. japonicum SjG28 gene fragment was selected as the target gene fragment, and the primers and fluorescent probe were designed and synthesized. Then, a S. japonicum nucleic acid dipstick test was established. The sensitivity of this dipstick test was evaluated by detecting different copies of recombinant plasmids containing the S. japonicum SjG28 gene fragment and different concentrations of genomic DNA from adult worms of S. japonicum, and the specificity of the dipstick test was evaluated by detecting the genomic DNA from Clonorchis sinensis, S. mansoni, Ancylostoma duodenale, S. haematobium, Babesia and Paragonimus westermani. Results　The S. japonicum nucleic acid dipstick test based on the S. japonicum SjG28 gene fragment showed the minimum detectable limit of 10 copies/μL of the recombinant plasmid containing the S. japonicum SjG28 gene fragment and the minimum detectable limit of 1 pg/μL of S. japonicum genomic DNA, and the dipstick assay tested negative for the genomic DNA from C. sinensis, S. mansoni, A. duodenale, S. haematobium, Babesia and P. westermani. Conclusion　A rapid, simple, and visualized assay is established for detection of S. japonicum specific gene fragments based on RAA and nucleic acid dipstick test.

Key words: Schistosoma japonicum, Nucleic acid dipstick, Recombinase?aided amplification, Nucleic acid detection, SjG28 gene

摘要： 目的　结合重组酶介导核酸等温扩增技术（recombinase⁃aided isothermal amplification assay，RAA）和核酸试纸条建立一种快速检测日本血吸虫特异性基因片段的方法。方法　以日本血吸虫SjG28基因片段作为靶序列，设计并合成引物、荧光探针，建立日本血吸虫核酸试纸条检测方法。通过检测不同拷贝数的含SjG28基因片段的重组质粒和不同浓度的日本血吸虫成虫基因组DNA，对该方法敏感性进行评价；通过检测华支睾吸虫、曼氏血吸虫、十二指肠钩口线虫、埃及血吸虫、巴贝斯虫和卫氏并殖吸虫基因组DNA，对该方法特异性进行评价。结果　以重组质粒为模板，建立的日本血吸虫特异性基因片段核酸试纸条检测方法最低检测限为10拷贝/μL；以成虫基因组DNA为模板，最低检测限为1 pg/μL。该方法检测华支睾吸虫、曼氏血吸虫、十二指肠钩口线虫、埃及血吸虫、巴贝斯虫和卫氏并殖吸虫基因组DNA结果均为阴性。结论　本研究结合RAA技术和核酸试纸条建立了一种可快速、简便、可视化检测日本血吸虫特异性基因片段的核酸试纸条法。

关键词: 日本血吸虫, 核酸试纸条, 重组酶介导的等温扩增, 核酸检测, SjG28基因