HLA‐AB typing by polymerase‐chain reaction with sequence‐specific primers: more accurate, less errors, and increased resolution compared to serological typing

Abstract

Until recently, serological typing has been the primary technique used for HLA class I analysis. But because of limitations, molecular-typing techniques have replaced or supplemented the microlymphocytotoxicity test. It has been assumed that HLA class I serological typing was more accurate than serological HLA-DR typing; the latter has been shown to have 10-25% errors. But several studies have shown that HLA-AB typing was poorer than expected, and error frequencies between 5-25% were reported. This study systematically investigated the accuracy of HLA class I serological AB typing in healthy, bone-marrow registry donors, necrokidney donors, kidney-transplantation patients (on waiting lists), and haematological disorder patients. Genomic HLA class I typing, which uses polymerase-chain reaction with sequence-specific primers (PCR-SSP), gave discrepant results in 3-24% of the patients, compared to serological typings. The highest error rate (24%) was found among haematological disorder patients. Among the kidney waiting-list patients and necrokidney donors, 11% discrepancies were found. In the consecutively typed bone-marrow donors group, 3% errors were found. But among those with only one detected HLA-A specificity, 12% discrepancies were found, and among donors with only one detected HLA-B specificity, 19% errors were found. Based on these results, we recommend that patients with haematological disorders should be typed using genomic techniques. In investigations of bone-marrow registry donors and kidney patients, in which only one serological specificity is found, additional typing by genomic methods should be done.

Quality control of a national bone marrow donor registry: results of a pilot study and proposal for a standardized approach

“…This estimate is very close to that reported for the Swedish Registry. 10 As also observed by previous investigators, [9][10][11][12] the missed antigens belong preferentially to the A19 and B70 groups and are notoriously difficult to define because of lack of monospecific antisera. The DNA retyping results therefore show that systematic verification by molecular typing of HLA-AB homozygous samples will significantly improve the histocompatibility records of the Registry.…”

“…9 More recently, a combined serology/polymerase chain reaction-sequence specific primers (PCR-SSP) analysis of donors from the Tobias Registry (Sweden) showed an overall 3% discrepancy rate, but among homozygotes, 12% for HLA-A and 19% for HLA-B discrepancies, respectively, were observed. 11 The largest study group of donors typed both by serology (performed in 1993-1997) and by PCR-SSOP hybridization or PCR-SSP concerns 440 000 non-Caucasoid donors from National Marrow Donor Program (NMDP). 12 An overall discrepancy rate of 24% has been reported, 8% for HLA-A, 13% for HLA-B, and 3% for both HLA-A and -B.…”

Immunogenetic Heterogeneity in Single-System and Multisystem Langerhans Cell Histiocytosis

“…The polymorphism detected with PCR-based typing methods correlates well with the polymorphism recognized by serology and cellular typing (27). More importantly, with DNA-based tissue typing, more biologically relevant polymorphisms may be detected with error frequencies lower than with serology (28,29).…”

Cyclosporine and mycophenolate mofetil prophylaxis with fludarabine and melphalan conditioning for unrelated donor transplantation: a prospective study of 22 patients with hematologic malignancies

“…Donor/recipient matching and stem cell source High-resolution polymerase chain reaction-sequencespecific-primers (PCR-SSP) for HLA class I and II was performed. 7 Donor/recipients were fully matched at HLA A, B, C, and DRB1 in 17 pairs, while five had HLA class I allele mismatches. Stem cell source, which was determined by the NMDP donor center, was bone marrow (n ¼ 11) and peripheral blood (n ¼ 11).…”