纳米磁珠联合PCR检测甘蔗杆状病毒方法的建立和初步应用

本研究通过对DNA提取裂解液的成分和用量以及裂解时间等因素进行系统优化,建立了一种高通量的磁珠法提取甘蔗总DNA,再进行PCR检测甘蔗杆状病毒(SCBV)的方法。采用核酸自动提取仪,应用纳米磁珠提取甘蔗总DNA,经PCR检测甘蔗杆状病毒,并与以试剂盒提取DNA为模板的PCR检测结果进行比较。结果显示,经优化的裂解液成分为EDTA·Na20.08 mol/L、NaCl 0.8 mol/L、SDS 2%和Tris-HCl0.10 mol/L,裂解时间为20 min,可用于纳米磁珠提取甘蔗总DNA进行SCBV检测。所建立的方法通量高、成本低、耗时短,可用于SCBV的批量检测。

Establishment and Preliminary Application of Nanomagnetic Beads Combined with PCR for Detecting Sugarcane Baculovirus

In this study,the composition,dosage and pyrolysis time of DNA extraction lysate were systematically optimized,and a high flux magnetic beads method for extracting total DNA from sugarcane as well as detecting Sugarcane bacilliform virus(SCBV)by PCR was established.Total DNA from sugarcane was extracted by nanomagnetic beads with automatic nucleic acid extraction instrument and SCBV was detected by PCR as well.The results were compared with those of PCR extracted from sugarcane with kit as template.Using an automatic nucleic acid extractor and the nano magnetic beads to extract the total DNA of sugarcane.After detection of Sugarcane bacilliform virus by PCR,the results of PCR detection were compared with those of DNA extraction using kit as template.The results showed that the components of the optimized lysate are EDTA·Na20.08 mol/L,NaCl0.8 mol/L,SDS 2%and Tris-HCl 0.10 mol/L,and the lysis time is 20 min.It can be used to extract total DNA from sugarcane by nano magnetic beads for SCBV detection,which is not only high throughput,but also low cost and time-saving.