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中文摘要：
为确定狂犬病病毒（RV）Flury-LEP株外源基因的插入位点及其中和抗体快速检测方法,本研究在建立了RVFlury-LEP株反向遗传操作系统的基础上,构建了表达增强型绿色荧光蛋白（EGFP）的重组LEP基因组cDNA克隆pCI-LEP-EGFP,并成功拯救出重组病毒（rLEP-EGFP）。rLEP-EGFP的生物学特性和野生型LEP（wtLEP）在NA细胞和BHK-21细胞中的生长动力学及对小鼠的致病性没有显著差异。rLEP-EGFP在BHK-21细胞中连续传代9次,各代次重组病毒均稳定表达EGFP。重组表达的EGFP可用于中和抗体检测,rLEP-EGFP免疫犬血清经间接免疫荧光（IFA）或荧光下直接观察检测RV中和抗体滴度,两种检测结果显示较好的相似性（p〉0.05）。本研究为研制以Flury-LEP株活载体多价疫苗奠定了基础,并为病毒中和抗体的检测提供了更为快捷的新方法。
英文摘要：
To construct the recombinant rabies virus and develop a convenient method for virus neutralization antibodies （VNA） detection, a recombinant rabies virus rLEP-EGFP expressing enhanced green fluorescent protein （EGFP） was rescued based on infectious clone of rabies virus Flury-LEP strain. The biology characters of rLEP-EGFP were similar to that of the wild type LEP strain in respect of the growth dynamics in both NA cells and BHK-21 cells, and the pathogenicity in BALB/c mice. The EGFP expression of rLEP-EGFP was stable for at least nine passages in BHK-21 cells. The rLEP-EGFP could be used for detection of neutralization antibody in canine serum, and the results were as same as that of indirect immunofluorescence assay （p0.05）. This LEP-EGFP system provided an useful platform for development of novel polyvalent vaccine, and a convenient and economical method for VNA detection.
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