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首页> 外文期刊>Transfusion: The Journal of the American Association of Blood Banks >B-lymphoblastoid cell lines as a source of reference DNA for human platelet and neutrophil antigen genotyping.
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B-lymphoblastoid cell lines as a source of reference DNA for human platelet and neutrophil antigen genotyping.

机译：B淋巴母细胞系作为人类血小板和中性粒细胞抗原基因分型的参考DNA来源。

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摘要

BACKGROUND: Human platelet and neutrophil antigens (HPAs, HNAs) are targets for platelet or granulocyte antibodies causing immune thrombocytopenia or neutropenia, respectively. Currently, genotyping is replacing phenotyping as the preferred method of diagnosis of immune cytopenia. To establish a reliable genotyping analysis, however, the availability as reference DNA of genomic DNA from persons of known genotype is essential. STUDY DESIGN AND METHODS: By the use of Epstein-Barr virus transformation, panels of B-lympho-blastoid cell lines (B-LCLs) from HPA- and HNA-phenotyped individuals were developed. Genomic DNA was isolated from these cell lines and tested as reference DNA for genotyping of persons for HPAs and HNAs. RESULTS: DNA derived from these B-LCLs was typed by polymerase chain reaction-restriction fragment length polymorphism and -sequence-specific primers. The results were in accordance with the genotyping from peripheral blood cells. These results were confirmed by 24 laboratories in Germany in a blind study. CONCLUSION: The inexhaustible source of reference DNA derived from B-LCLs allowed the evaluation of reliable HPA and HNA genotyping for quality control purposes. It should facilitate the development of DNA typing in blood centers and clinical laboratories.
机译：背景：人类血小板和嗜中性粒细胞抗原（HPA，HNA）分别是引起免疫性血小板减少或中性粒细胞减少的血小板或粒细胞抗体的靶标。目前，基因分型正在取代表型，成为诊断免疫性血细胞减少症的首选方法。然而，为了建立可靠的基因分型分析，从已知基因型的人获得基因组DNA作为参考DNA是必不可少的。研究设计和方法：通过使用爱泼斯坦-巴尔病毒转化，开发了来自HPA和HNA表型个体的B淋巴母细胞细胞系（B-LCL）。从这些细胞系中分离出基因组DNA，并将其作为参考DNA进行人的HPA和HNA基因分型。结果：通过聚合酶链反应-限制性片段长度多态性和-序列特异性引物对源自这些B-LCL的DNA进行分型。结果与外周血细胞的基因分型一致。这些结果在德国的24个实验室进行了盲目研究中得到了证实。结论：来自B-LCL的参考DNA的取之不尽用之不竭，因此可以评估出于质量控制目的的可靠HPA和HNA基因分型。它应该促进血液中心和临床实验室中DNA分型的发展。

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