通过双标记时间分辨免疫荧光分析法同时定量两种植物病毒。
Simultaneous quantification of two plant viruses by double-label time-resolved immunofluorometric assay.
作者信息
Saarma M, Järvekülg L, Hemmilä I, Siitari H, Sinijärv R
机构信息
Laboratory of Molecular Genetics, Estonian Academy of Sciences, U.S.S.R.
出版信息
J Virol Methods. 1989 Jan;23(1):47-54. doi: 10.1016/0166-0934(89)90088-8.
For simultaneous and sensitive detection of two antigens in one sample, monoclonal antibody (MAb) to potato virus M (PVM) was labelled with a lanthanide Eu3+ and MAb to potato virus X (PVX) with another lanthanide Sm3+. A mixture of the labelled MAbs was used as a tracer. After performing the immunoreactions, the fluorescence of the dissociated lanthanides was measured at different wave-lengths in a time-resolved fluorometer to quantificate the PVX and PVM amount in a sample. Double-label time-resolved fluoroimmunoassay (TR-FIA) detected 1 ng/ml of each virus and was therefore more sensitive for simultaneous detection of PVX and PVM than reported for a single virus detection with double antibody sandwich ELISA (DAS-ELISA).
为了在一个样本中同时灵敏地检测两种抗原，用镧系元素铕（Eu3+）标记马铃薯M病毒（PVM）的单克隆抗体（MAb），用另一种镧系元素钐（Sm3+）标记马铃薯X病毒（PVX）的单克隆抗体。标记的单克隆抗体混合物用作示踪剂。进行免疫反应后，在时间分辨荧光计中于不同波长测量解离的镧系元素的荧光，以定量样本中PVX和PVM的含量。双标记时间分辨荧光免疫分析（TR-FIA）可检测到每种病毒1 ng/ml，因此与用双抗体夹心ELISA（DAS-ELISA）进行的单一病毒检测相比，对同时检测PVX和PVM更灵敏。
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