Large-scale DNA typing for human platelet alloantigens by PCR-PHFA (preferential homoduplex formation assay)

被引:27
作者
Fujiwara, K
Isa, K
Oka, T
Maekawajiri, S
Yamane, A
Akaza, T
Tadokoro, K
Juji, T
Shibata, Y
Tokunaga, K
机构
[1] WAKUNAGA PHARMACEUT CO LTD,BIOTECHNOL RES INST,HIROSHIMA,JAPAN
[2] UNIV TOKYO,FAC MED,DEPT TRANSFUS MED & IMMUNOHAEMATOL,TOKYO 113,JAPAN
[3] UNIV TOKYO,SCH INT HLTH,DEPT HUMAN GENET,TOKYO,JAPAN
关键词
human platelet alloantigens; DNA typing; single base substitution; preferential homoduplex formation assay;
D O I
10.1046/j.1365-2141.1996.d01-1871.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Alloimmunization against human platelet alloantigens (TTPA) is known to be involved in disorders such as neonatal alloimmune thrombocytopenic purpura, posttransfusion purpura, and refractoriness to platelet transfusion therapy. HPA typing is essential in diagnosis and management of patients. Therefore a reliable and speedy method is necessary for HPA typing. We have successfully applied a new DNA typing method, PCR-preferential homoduplex formation assay (PHFA) method, to typing for the HPA-1, -2, -3, -4, -5 and -6 systems. This method is based on DNA strand competition during hybridization under a precisely controlled temperature gradient between a double-labelled amplicon (standard DNA), prepared from biotin- and DNP-labelled primers, and an unlabelled amplicon (sample DNA). The results obtained by PCR-PHFA typing were in good agreement with the allotypes determined by serological typing and by other DNA typing methods. The PCR-PHFA method can be easily automated, is suitable for typing both small and large numbers of samples, and thus is applicable to routine HPA typing.
引用
收藏
页码:198 / 203
页数:6
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