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聚合酶链反应-序列特异性寡核苷酸探针技术(PCR-SSOP)与序列特异性引物分型技术(SBT)在人类白细胞抗原(HLA)分型中的差异:一项病例研究

Discrepancies in HLA typing by PCR-SSOP and SBT techniques: a case study.

作者信息

Spínola Hélder, Bruges-Armas Jácome, Brehm António

机构信息

Human Genetics Laboratory, University of Madeira, Campus of Penteada, 9000-399 Funchal, Portugal.

出版信息

Hum Biol. 2007 Oct;79(5):537-43. doi: 10.1353/hub.2008.0001.

Abstract

Six hundred twenty-one samples from Portugal, the Cabo Verde archipelago, and Guinea-Bissau were typed for HLA-A, HLA-B, and HLA-DRB1 using the polymerase chain reaction-sequence-specific oligonucleotide probe (PCR-SSOP) method and the sequence-based typing (SBT) method to characterize and compare discrepancies between the two methods. Fifty-three alleles (4.27% of 1242 chromosomes typed) identified by the PCR-SSOP method were not concordant with the results obtained using the SBT method. Thirty-four (2.74% of total chromosomes typed) PCR-SSOP mistyping results were discrepancies inside the same allele group and 19 others (1.53% of total chromosomes typed) were relative to nonconcordant results between different groups. PCR-SSOP allele mistyping is the result of interpretation difficulties resulting from less intense, absent, or dubious hybridization patterns. Noncommercial PCR-SSOP procedures are highly exigent on the technicians' experience and the availability of properly calibrated high-precision equipment.

摘要

使用聚合酶链反应-序列特异性寡核苷酸探针(PCR-SSOP)方法和基于序列的分型(SBT)方法,对来自葡萄牙、佛得角群岛和几内亚比绍的621个样本进行HLA-A、HLA-B和HLA-DRB1分型,以表征和比较两种方法之间的差异。PCR-SSOP方法鉴定出的53个等位基因(占1242条分型染色体的4.27%)与使用SBT方法获得的结果不一致。34个(占总分型染色体的2.74%)PCR-SSOP错误分型结果是同一等位基因组内的差异,另外19个(占总分型染色体的1.53%)与不同组之间的不一致结果有关。PCR-SSOP等位基因错误分型是由杂交模式强度较低、缺失或可疑导致的解读困难造成的。非商业PCR-SSOP程序对技术人员的经验以及校准良好的高精度设备的可用性要求很高。

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