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淋巴细胞脉络丛脑膜炎病毒实时荧光RPA快速检测方法的建立

点击数:6689  时间:2019-06-26 10:31:37  来源: 信息处      

针对淋巴细胞脉络丛脑膜炎病毒(lymphocytic choriomeningitis virus,LCMV)基因保守序列,设计特异性引物和荧光探针,建立了LCMV实时重组聚合酶等温扩增(real-time recombinasepolymerase amplification,real-time RPA)检测方法。该检测方法具有良好的特异性,与LCMV同步检测的其他鼠病毒均未出现交叉反应;该检测方法具有与RT-PCR一样的高敏感性;通过对LCMV感染鼠组织样本和LCMV阴性鼠血清样本的检测,证实建立的实时荧光RPA方法具有良好的稳定性和可靠性。与现有的核酸检测方法相比,该实时荧光RPA检测方法在核酸上样20 min内即可判读结果,可满足啮齿类动物LCMV快速检疫需要。



Establishment of a Real-time RPA Method for Rapid Detection ofLymphocytic Choriomeningitis Virus

In this paper,specificprimers and fluorescent probes were designed according to the conservativesequence of lymphocytic choriomeningitis virus(LCMV),and a real-time recombinase polymerase amplification assay(real-time RPA)was established. Resultsshowed that the specificity of the assay was good,andno any cross reaction with other murine viruses detected by LCMV was shown. Thesensitivity of established assay was as high as the RT-PCR method. Otherwise,based on detection of LCMV infected murine tissues and negativeserums,the real-time RPA showed good stability andreliability. Compared with current detection methods of nucleic acids,the test results could be interpreted within 20 minutes after loadingthe nucleic buffer using the real-time RPA,which couldsatisfy the need of rapid LCMV inspection in rodent animals.

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