2.2.15细胞上清HBVDNA定量检测方法的比较

Quantitiative detection HBVDNA in culture media of 2.2.15 cell

目的拟筛选和比较2.2.15细胞上清中HBVDNA定量检测的方法。方法分别采用3种方法抽提HBVDNA,抽提物分别采用Taqman荧光定量、SYBR荧光定量PCR技术检测其滴度。结果直接应用上清分别采取热变性,碱裂解法抽提HBVDNA均为阴性,而应用PEG沉淀后检测结果证实细胞上清中有高拷贝的分泌性HBVDNA,Taqman荧光定量方法的敏感性高于SYBR实时定量PCR,在高拷贝时,两种方法的检测结果一致。结论PEG沉淀法是2.2.15细胞上清HBVDNA提取的有效方法,Taqman荧光定量方法敏感性高,是HBVDNA定量检查的首选方法。SYBR法简便,经济,也可用于上清HBVDNA的检测。

TO explore the methods of quantitiative detection secretary HBVDNA of 2.2.15 cell. The HBVDNA in culture media were prepared with three methods for isolation and amplified by Taq man and SYBR real-time quatitiative PCR. HBVDNA could not detected by the direct prepratation of culture media. PEG precipitate method was shown to have high copy of secretary HBVDNA in culture media of 2.2.15 cell. The sensitivity of Taq man real time PCR was better than SYBR real-time quantitiative PCR. The results of detetion by two methods was the same when the concentration of HBVDNA was high. [Conclusion] The PEG precipitate method is the pratical method of HBVDNA isolation. Taq man real time PCR is the first -choise of HBVDNA quantitiative analysis. SRBR real time PCR is a simple, economical method of HBVDNA quantitiative detetion.