基于重组酶聚合酶扩增结合侧流层析技术的B、E型腺病毒快速可视化检测方法的研究与评价

发布时间：2018-06-20 05:15

  本文选题：腺病毒科 + 重组酶聚合酶扩增技术　； 参考：《军事医学》2017年07期

【摘要】：目的建立一种快速、准确、可视化、易开展的B、E型腺病毒检测技术。方法针对B、E型腺病毒壳蛋白保守序列设计通用引物,建立可同时检测出这两种腺病毒型别的重组酶聚合酶扩增结合侧流层析试纸条(recombinase polymerase amplification combined with a lateral flow dipstick,LFD-RPA)检测技术,评价其灵敏度与特异性,并验证最佳反应温度与反应时间,同时利用该法对19例B、E型腺病毒感染患者及10例健康志愿者鼻咽拭子标本进行检测。结果腺病毒LFD-RPA法检测灵敏度可达10拷贝/μl,与q PCR方法相近,且不会与其他亚属腺病毒及其他病毒发生交叉反应。反应可在25~45℃温度范围内高效进行,检测全程仅需15~20 min。利用临床咽拭子标本对检测体系进行评估,其灵敏度及特异性均达100%。结论该检测方法灵敏度高、特异性强、操作简单,反应快速,能脱离对大型仪器、专业操作人员及正规实验室的依赖,在基层卫生部门,尤其在野外及现场检测中具有极大的应用潜力。
[Abstract]:Objective to establish a rapid, accurate, visual and easy detection technique for adenovirus type E (AV). Methods A universal primer was designed for the conserved sequence of adenovirus type E adenovirus shell protein, and a recombinant enzyme polymerase amplification method combined with LFD-RPAA assay was developed to detect the two adenovirus types simultaneously. The recombinant polymerase amplification combined with side flow chromatography test strip was used to detect the recombinant adenovirus types. The sensitivity and specificity were evaluated, and the optimum reaction temperature and reaction time were verified. At the same time, the nasopharyngeal swabs of 19 patients with adenovirus type E infection and 10 healthy volunteers were detected by this method. Results the sensitivity of LFD-RPA assay was up to 10 copies / 渭 l, which was similar to that of Q PCR, and no cross reaction with other subgenus adenovirus and other viruses was observed. The reaction can be carried out efficiently in the temperature range of 25 鈩，

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