Discrepancies in HLA Typing by PCR-SSOP and SBT Techniques: a case study

Six hundred twenty-one samples from Portugal, the Cabo Verde archipelago, and Guinea-Bissau were typed for HLA-A, HLA-B, and HLADRB1usingthepolymerasechainreaction–sequence-specificoligonucleotide probe (PCR-SSOP) method and the sequence-based typing (SBT) method to characterizeandcomparediscrepancie...

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Bibliographic Details
Main Author: Spínola, Hélder (author)
Other Authors: Bruges-Armas, Jácome (author), Brehm, António (author)
Format: article
Language:eng
Published: 2017
Subjects:
Online Access:http://hdl.handle.net/10400.13/1351
Country:Portugal
Oai:oai:digituma.uma.pt:10400.13/1351
Description
Summary:Six hundred twenty-one samples from Portugal, the Cabo Verde archipelago, and Guinea-Bissau were typed for HLA-A, HLA-B, and HLADRB1usingthepolymerasechainreaction–sequence-specificoligonucleotide probe (PCR-SSOP) method and the sequence-based typing (SBT) method to characterizeandcomparediscrepanciesbetweenthetwomethods.Fifty-three alleles (4.27% of 1,242 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.