針對(duì)淋巴細(xì)胞脈絡(luò)叢腦膜炎病毒（lymphocytic choriomeningitis virus，LCMV）基因保守序列，設(shè)計(jì)特異性引物和熒光探針，建立了LCMV實(shí)時(shí)重組聚合酶等溫?cái)U(kuò)增（real-time recombinasepolymerase amplification，real-time RPA）檢測(cè)方法。該檢測(cè)方法具有良好的特異性，與LCMV同步檢測(cè)的其他鼠病毒均未出現(xiàn)交叉反應(yīng)；該檢測(cè)方法具有與RT-PCR一樣的高敏感性；通過對(duì)LCMV感染鼠組織樣本和LCMV陰性鼠血清樣本的檢測(cè)，證實(shí)建立的實(shí)時(shí)熒光RPA方法具有良好的穩(wěn)定性和可靠性。與現(xiàn)有的核酸檢測(cè)方法相比，該實(shí)時(shí)熒光RPA檢測(cè)方法在核酸上樣20 min內(nèi)即可判讀結(jié)果，可滿足嚙齒類動(dòng)物L(fēng)CMV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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