高梯度磁性细胞分选和内标法显著改进了对血型糖蛋白A（GPA）位点体细胞突变的检测。

High gradient magnetic cell sorting and internal standardisation substantially improve the assay for somatic mutations at the glycophorin A (GPA) locus.

作者信息

Hempel Klaus, Deubel Werner, Lorenz Reinhard, Reiners Christoph

机构信息

Klinik und Poliklinik für Nuklearmedizin der Universität, Würzburg, Josef-Schneider-Str 2, D-97080 Würzburg, Germany.

出版信息

Mutat Res. 2003 Apr 9;525(1-2):29-42. doi: 10.1016/s0027-5107(02)00311-1.

DOI:10.1016/s0027-5107(02)00311-1

PMID:12650903

Abstract

In the MACS-BR6 version of the GPA assay [Int. J. Radiat. Res. 70 (1996) 131] variant red blood cells (RBC) are isolated from 5 x 10(8) normal RBC by magnetic cell separation (MACS) before detection and quantification by immunolabelling and flow cytometry as in the classical BR6 assay. In the present work it is described how the MACS-BR6 assay is improved by internal standardisation with FITC-labelled RBC. This modification of the assay has the advantage that (i) the analysis of variants is not disturbed by the overwhelming number of normal RBC that (ii) the precision of the assay is improved and finally that (iii) a sufficient number of variants is available for further investigations. Tn positive RBC behave in MACS like variants. It is demonstrated that in normal individuals Tn cells (frequency: approximately 2 x 10(-8)) do not disturb the assay.

摘要

在GPA分析的MACS - BR6版本中[《国际辐射研究杂志》70 (1996) 131]，在通过免疫标记和流式细胞术进行检测和定量之前，通过磁珠细胞分选（MACS）从5×10⁸个正常红细胞中分离出变异红细胞（RBC），如同经典的BR6分析一样。在本研究中，描述了如何通过用异硫氰酸荧光素（FITC）标记的红细胞进行内部标准化来改进MACS - BR6分析。该分析方法的这种改进具有以下优点：（i）变异体的分析不会受到大量正常红细胞的干扰；（ii）分析的精度得到提高；最后（iii）有足够数量的变异体可用于进一步研究。Tn阳性红细胞在MACS中的行为类似于变异体。结果表明，在正常个体中，Tn细胞（频率：约2×10⁻⁸）不会干扰该分析。