-D- phenotype screening by polybrene method
10.13303/j.cjbt.issn.1004-549x.2021.11.003
- VernacularTitle:应用聚凝胺筛选-D-表型红细胞的研究
- Author:
Fengyong ZHAO
1
;
Zhongying WANG
1
;
Yuyu ZHANG
1
;
Qin LI
1
;
Yunxiang WU
2
;
Fei WANG
2
;
Qing LI
2
;
Demei ZHANG
2
;
Dong XIANG
1
;
Ziyan ZHU
1
Author Information
1. Shanghai Blood Center, Shanghai 200051, China
2. Taiyuan Blood Center
- Publication Type:Journal Article
- Keywords:
-D-phenotype;
polybrene;
RhD-- gene carrier;
RhD antigen expression;
red blood cell
- From:
Chinese Journal of Blood Transfusion
2021;34(11):1186-1190
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.
