利用荧光定量PCR检测猪细小病毒在IBRS-2细胞增殖动态研究

Study on the Proliferation Dynamics of Porcine Parvovirus in IBRS-2 Cells Using Real-time PCR Technology

为了解猪细小病毒(PPV)在IBRS-2细胞中的增殖规律,建立了基于TaqMan实时荧光定量PCR技术的PPV检测方法,并应用该方法研究PPV在IBRS-2细胞中的增殖动态。结果显示,该方法特异性较好且灵敏度高,可检测低至1.4×100拷贝/μL的病毒量。检测显示PPV接种IBRS-2细胞后12h开始大量增殖,60～72h增殖最为迅速,到72h病毒含量达到最高峰,之后逐渐下降。结果表明,所建立的荧光定量PCR方法可用于PPV的快速定量检测,对病毒增殖规律的研究为该病毒疫苗的生产提供了科学依据。

To explore the proliferation dynamics of PPV in IBRS-2 cells,a TaqMan fluorescent quantitative real-time PCR technology was established. The results showed that this method is specific and sensitive for the detection of PPV. Experiment of sensitivity indicated that this method could detect 1.4？100copies/μL viral load at least. After inoculation of PPV, the IBRS-2 cells showed obvious CPE. Virus began to proliferate when the cells infected with PPV after 12 hours, and proliferated rapidly during 60 to 72 hours, viral load achieved the highest value in 72hours and then decreased gradually.This study results indicated that the TaqMan Real-time PCR technology could be used in the rapid quantitative detection of PPV. The proliferation dynamics also could provided a scientific basis for the vaccine production of PPV.