为建立一种灵敏、可快速检测猪圆环病毒4型（PCV4）的SYBR Green I实时荧光定量PCR方法，根据PCV4 Rep基因的保守区域设计引物，建立针对PCV4的SYBR Green I实时荧光定量PCR检测方法，并对该方法的特异性、灵敏性和重复性进行评价。结果显示，该检测方法的Ct值与标准品在5.64×102~5.64×109 copies/μL范围内呈良好的线性关系，R2为0.999，斜率为-3.638，检测下限为5.64×102 copies/μL，且无非特异性扩增。利用该方法对56份临床样品进行检测，发现PCV4核酸阳性率为3.57%，比普通PCR更灵敏可靠。结果表明，本试验建立的SYBR Green I实时荧光定量PCR方法可用于PCV4的快速诊断。
In order to rapidly and sensitively detect porcine circovirus type 4（PCV4），the SYBR Green I real-time quantitative PCR assay was established based on the primers that were designed according to the conserved region of PCV4 Rep gene. Then its specificity，sensitivity and repeatability were evaluated. The results showed that its Ct value remained a good linear relationship with the standard materials within the range of 5.64×109~5.64×102 copies/μL，specifically，R2 was 0.999，the slope was -3.638，and the detection limit was 5.64×102 copies/μL without non-specific amplification. 56 clinical samples were tested by the method，it was found that the nucleic acid positive rate of PCV4 was 3.57%，which was more sensitive and reliable than that of conventional PCR. Therefore，the established SYBR Green I real-time PCR method could achieve the rapid detection of PCV4.
黄小武,杜倩,闭璟珊,韦正吉,邹联斌,赵子欣,闫修魁,韩知晓,辛佳亮,郑敏. PCV4 SYBR Green I实时荧光定量PCR方法的建立及应用[J].《中国动物检疫》编辑部,2020,37(10):109-113.复制