Donor HLA Genotyping using Polymerase Chain Reaction- Sequence Specific Primers (PCR-SSP) as Method for Acquiring Donor Panel in Platelet Refractoriness

T. D. Atmakusuma, A. Rachman, N. K. Ritchie
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Abstract

Background: Evaluation and identification of HLA antibodies in the recipient’s serum is of utmost importance prior to transplantation and transfusion. HLA typing is a steppingstone in proposing a donor panel. In order to obtain the HLA typing, Polymerase Chain Reaction-Sequence Specific Primers (PCR-SSP) can be performed.Materials and method: This is a preliminary study to determine HLA polymorphism by HLA genotyping in 43 blood donors. DNA from the samples was isolated using commercial kits according to the standard protocol. The DNA then was amplified using PCR-SSP methods and analyzed using the provided set in the kit.Results: This study found that the most frequent HLA-A alleles was HLA-A*24 (41.9%). For HLA-B alleles, the most common was HLA-B*15 (28%). Most frequent HLA-A-B haplotypes was HLA-A*24-B*15 (11.3%). The results from this study concurs with that of previous study. However, some alleles might vary due to difference in study population. Determining HLA-typing is of paramount importance in an ethnically diverse country such as Indonesia. In contrast to homogenous caucassian country, difference in ethnicity might cause platelet refractoriness due to incompatibility. HLA-typing would also guide the diagnostic workup and required treatment strategy for platelet refractoriness.Conclusion: From the HLA typing using PCR-SSP in blood donors in Jakarta, we found that the most frequent alleles were HLA-A*24 and HLA-B*15; and the most frequent haplotypes were HLA-A*24-B*15. This study should be upscaled to include larger population and ethnic groups to obtain complete profile of Indonesian population.Keywords: platelet refractoriness, HLA, donor, PCR-SSP, transfusion medicine
利用聚合酶链反应-序列特异性引物(PCR-SSP)进行供体HLA基因分型以获得血小板难耐性供体样本
背景:在移植和输血前,受体血清HLA抗体的评估和鉴定是至关重要的。HLA分型是建立供体小组的基础。为了获得HLA分型,可以使用聚合酶链反应-序列特异性引物(PCR-SSP)。材料与方法:采用HLA基因分型法测定43例献血者HLA多态性的初步研究。根据标准方案使用商业试剂盒从样品中分离DNA。然后使用PCR-SSP方法扩增DNA,并使用试剂盒中提供的集合进行分析。结果:本研究发现HLA-A等位基因最常见的是HLA-A*24(41.9%)。HLA-B等位基因最常见的是HLA-B*15(28%)。最常见的HLA-A- b单倍型为HLA-A*24-B*15(11.3%)。这项研究的结果与以前的研究结果一致。然而,由于研究人群的不同,一些等位基因可能会有所不同。确定hla分型在印度尼西亚这样一个种族多样化的国家至关重要。与单一的高加索国家相比,种族差异可能导致血小板不相容。hla分型也将指导诊断工作和血小板难治性所需的治疗策略。结论:利用PCR-SSP对雅加达献血者进行HLA分型,发现HLA- a *24和HLA- b *15是最常见的等位基因;最常见的单倍型为HLA-A*24-B*15。这项研究应该扩大到包括更多的人口和种族群体,以获得印度尼西亚人口的完整概况。关键词:血小板难固性,HLA,供体,PCR-SSP,输血医学
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