One-tube method for complete HPA-1 genotyping by PCR using sequence-specific primers

被引:4
作者
Boldt, B
Skogen, B
Agostini, T
Roscetti, D
McFarland, J
机构
[1] Genet Testing Inst, Brookfield, WI 53045 USA
[2] Univ Tromso Hosp, N-9012 Tromso, Norway
[3] Blood Ctr SW Wisconsin, Milwaukee, WI USA
关键词
human platelet antigen; HPA-1; neonatal alloimmune thrombocytopenia; genotyping; PCR;
D O I
10.1046/j.1365-2141.1997.5013303.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Human platelet antigens (HPA) can be targets for antibody responses that cause life-threatening thrombocytopenia following platelet transfusions or pregnancy As an aid to diagnosis and prevention, serologic and DNA-based methods have been developed to type HPA. Of the DNA-based strategies, those using the polymerase chain reaction (PCR) are very sensitive, but often require processing of amplification products. Sequence-specific primers (SSP) in the PCR eliminate the need for extensive handling of reaction products beyond gel electrophoresis. However, current methods require a separate reaction for each allele being typed. In this report we describe a method to simultaneously and completely genotype both alleles of HPA-1 in a single PCR. In addition, because the absence of an amplification product might also show the failure of a SSP, we introduced a recombinant template that can only be amplified by the SSP, thus ensuring primer performance and the identified genotype.
引用
收藏
页码:968 / 973
页数:6
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