使用整体式色谱载体浓缩植物病毒。

Concentration of plant viruses using monolithic chromatographic supports.

作者信息

Kramberger Petra, Petrovic Natasa, Strancar Ales, Ravnikar Maja

机构信息

BIA Separations, d.o.o., Teslova 30, SI-1000 Ljubljana, Slovenia.

出版信息

J Virol Methods. 2004 Sep 1;120(1):51-7. doi: 10.1016/j.jviromet.2004.03.018.

DOI:10.1016/j.jviromet.2004.03.018

PMID:15234809

Abstract

A new chromatographic medium, Convective Interaction Media(CIM) disk monolithic columns, was applied to plant virus concentration. The ability of the columns to concentrate highly diluted plant viruses was tested on a model plant virus, rod-shaped tomato mosaic virus (ToMV). Enzyme-linked immunosorbent assay (ELISA) was used for the quantitative analysis. The virus was concentrated using a strong anion exchanger, CIM quaternary amine (QA) disk monolithic column. A high salt concentration was used to elute the concentrated virus from the columns. It has been demonstrated that ToMV, which had been diluted considerably below the sensitivity of ELISA, was concentrated by several orders of magnitude in the one-step procedure. Concentrated virus preparations could be used directly for ELISA testing. In comparison with methods described for concentrating plant viruses from irrigation water, the above procedure may provide a much faster and more efficient way to concentrate highly diluted plant viruses. The procedure could be applied to the testing of other highly diluted plant viruses, and to concentrating viruses for antiserum production.

摘要

一种新型色谱介质——对流相互作用介质（CIM）盘状整体柱，被应用于植物病毒浓缩。在一种模型植物病毒——杆状番茄花叶病毒（ToMV）上测试了该柱浓缩高度稀释植物病毒的能力。采用酶联免疫吸附测定法（ELISA）进行定量分析。使用强阴离子交换剂CIM季胺（QA）盘状整体柱对病毒进行浓缩。采用高盐浓度从柱上洗脱浓缩的病毒。结果表明，在一步操作中，被稀释至远低于ELISA检测灵敏度的ToMV被浓缩了几个数量级。浓缩的病毒制剂可直接用于ELISA检测。与从灌溉水中浓缩植物病毒的方法相比，上述方法可能为浓缩高度稀释的植物病毒提供一种更快、更高效的途径。该方法可应用于其他高度稀释植物病毒的检测，以及用于生产抗血清的病毒浓缩。