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Development and Validation of a Fully Automated Platform for Extended Blood Group Genotyping

机译:扩展血型基因分型的全自动平台的开发和验证

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摘要

Thirty-five blood group systems, containing >300 antigens, are Listed by the International Society of Blood Transfusion. Most of these antigens result from a single nucleotide polymorphism. Blood group typing is conventionally performed by serology. However, this technique has some limitations and cannot respond to the growing demand of blood products typed for a Large number of antigens. The knowledge of the molecular basis of these red blood cell systems allowed the implementation of molecular biology methods in immunohematology laboratories. Here, we describe a blood group genotyping assay based on the use of TKL immobilization support and microarray-based HIFI technology that takes approximately 4 hours and 30 minutes from whole-blood samples to results analysis. Targets amplified by multiplex PCR were hybridized on the chip, and a revelation step allowed the simultaneous identification of up to 24 blood group antigens, Leading to the determination of extended genotypes. Two panels of multiplex PCR were developed: Panel 1 (KEL1/2, KEL3/4; JK1/2; FY1/2; MNS1/2, MNS3/4, FY*Fy et FY*X) and Panel 2 (YT1/2; C01/2; D01/2, HY+, Jo(a+); LU1/2; DI1/2). We present the results of the evaluation of our platform on a panel of 583 and 190 blood donor samples for Panel 1 and 2, respectively. Good correlations (99% to 100%) with reference were obtained.
机译:国际输血协会列出了包含> 300个抗原的三十五个血型系统。这些抗原大多数来自单核苷酸多态性。血型分型通常通过血清学进行。但是,该技术有一些局限性,不能满足对大量抗原分型的血液制品日益增长的需求。这些红细胞系统的分子基础知识使免疫血液学实验室可以实施分子生物学方法。在这里,我们描述了基于TKL固定化支持和基于微阵列的HIFI技术的血型基因分型测定,从全血样品到结果分析大约需要4小时30分钟。通过多重PCR扩增的靶标在芯片上杂交,揭示步骤可同时鉴定多达24种血型抗原,从而确定了扩展的基因型。开发了两个多重PCR面板:面板1(KEL1 / 2,KEL3 / 4; JK1 / 2; FY1 / 2; MNS1 / 2,MNS3 / 4,FY * Fy et FY * X)和面板2(YT1 / 2 ; C01 / 2; D01 / 2,HY +,Jo(a +); LU1 / 2; DI1 / 2)。我们分别在583个和190个献血者样本的面板1和面板2上展示了我们平台的评估结果。获得了与参考的良好相关性(99%至100%)。

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