【摘要】 本研究拟建立一种多重荧光PCR检测技术，可同时检测登革病毒（Dengue virus,DENV）、寨卡病毒（Zika virus,ZIKV）和西尼罗病毒（West Nile virus,WNV）。通过设计上述3种病原体的特异性引物，优化荧光PCR反应体系和扩增条件，再对多重荧光PCR反应体系的特异性、灵敏度和重复性进行验证。实验结果显示，建立的多重荧光PCR方法的引物和探针终浓度分别为600 nmol/L和400 nmol/L，退火温度为60℃；其最低检测限浓度为1.0×103拷贝/mL，具有较好的特异性和重复性，病原体之间无交叉反应。本研究所建立的多重荧光PCR检测方法可用于登革病毒、寨卡病毒和西尼罗病毒的定性检测，为蚊媒传染病的检测提供重要技术支持。更多还原

【Abstract】 In this study, a multiplex fluorescence PCR detection technique was established to simultaneously detect Dengue virus(DENV), Zika virus(ZIKV) and West Nile virus(WNV). Three pathogen-specific primers were designed by multiple PCR detection method, and the fluorescence PCR reaction system and amplification conditions were screened, and then the specificity, sensitivity and repeatability of the fluorescence PCR reaction system were verified. The final concentrations of primers and probes were 600 nmol/L and 400 nmol/L, respectively, and the annealing temperature was 60℃. The lowest detection limit was 1.0×103 copies/mL, with good specificity and reproducibility, and no cross-reactivity between pathogens. The multiplex PCR technique established in this study can be used for the qualitative detection of Dengue virus, Zika virus and West Nile virus, providing important technical support for the detection and diagnosis of mosquito-borne infectious diseases.更多还原