【摘要】 根据GenBank上鹅源新城疫病毒NA-1株M基因的序列,在保守区域设计并合成了1对引物,采用荧光嵌合法(SYBR GreenⅠ)建立了检测鹅源新城疫病毒的实时荧光定量PCR(real-time PCR)。以鹅源新城疫病毒NA-1株反转录产物cDNA为标准品,通过优化反应条件,建立了标准曲线,并进行了融解曲线分析。结果,标准曲线的Ct值检测范围为23~36,相关系数(r2)为0.992;无引物二聚体及非特异性产物,且Tm=(83±0)℃。结果表明,建立的检测鹅源新城疫病毒实时荧光定量PCR方法特异性强、灵敏性高,能为以后快速诊断鹅源新城疫病毒提供有效保障。更多还原

【Abstract】 According to the M gene sequence of Newcastle disease virus(NDV) strain NA-1 from goose available in GenBank,a pair of primers was designed for establishing a SYBR Green Ⅰreal-time PCR for M gene of NDV.To establish the standard curve,the cDNA of the NDV strain NA-1 was served as a stan-dard.The analysis of melting curve was also carried out.The results indicated that the linear range of Ct values ranged from 23 to 36 with a good correlation coefficient(r2=0.992) and the melting curve also showed a single peak with an Tm value of(83±0) ℃.It was concluded that the established real-time PCR assay was highly sensitive and specific,which could provide a tool for rapid diagnosis of NDV in clinical samples from geese.更多还原