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淋巴細胞脈絡叢腦膜炎病毒實時熒光RPA快速檢測方法的建立

點擊數:11914  時間:2019-06-26 10:31:37  來源: 信息處      

針對淋巴細胞脈絡叢腦膜炎病毒(lymphocytic choriomeningitis virus,LCMV)基因保守序列,設計特異性引物和熒光探針,建立了LCMV實時重組聚合酶等溫擴增(real-time recombinasepolymerase amplification,real-time RPA)檢測方法。該檢測方法具有良好的特異性,與LCMV同步檢測的其他鼠病毒均未出現交叉反應;該檢測方法具有與RT-PCR一樣的高敏感性;通過對LCMV感染鼠組織樣本和LCMV陰性鼠血清樣本的檢測,證實建立的實時熒光RPA方法具有良好的穩定性和可靠性。與現有的核酸檢測方法相比,該實時熒光RPA檢測方法在核酸上樣20 min内即可判讀結果,可滿足齧齒類動物LCMV快速檢疫需要。



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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