Large‐scale oligonucleotide typing for HLA‐DRB1/3/4 and HLA‐DQB1 is highly accurate, specific, and reliable

Abstract

DNA typing of HLA class II alleles of the DRB1/3/4 and DQB1 loci using sequence-specific oligonucleotide probes and polymerase chain reaction-amplified DNA was used for the large-scale typing of donors for the National Marrow Donor Program unrelated donor registry. The results of quality control analysis for the first 7 months of the project show the typing to be highly accurate, specific, and reliable. The percent of correctly classified HLA oligotypes based on 1652 DRB1 and 1652 DQB1 assignments was greater than 99% for DRB1/DRB3/DRB4 and greater than 98% for DQB1. This level of accuracy is particularly remarkable because the quality control samples could not be distinguished from 9011 donor samples tested at the same time by the laboratories.

Rapid high-throughput human leukocyte antigen typing by massively parallel pyrosequencing for high-resolution allele identification

“…Using this method, optimized high-resolution HLA genotyping would require haplotype separation for the reduction of ambiguities caused by the cis/trans difficulty, but can be achieved by group-specific polymerase chain reaction (PCR) [14] or haplotypes-specific extraction [15]. Developing and applying sequence-specific primer [16 -19] or sequence-specific oligonucleotide probes [20] are some of the solutions to resolve this ambiguity problem, which would otherwise require the sequencing of cloned DNA.…”

Estimation of HLA-A, -B, -DRB1 Haplotype Frequencies Using Mixed Resolution Data from a National Registry with Selective Retyping of Volunteers

“…Typing was done using a variety of techniques at varying levels of resolution (Table 1) as the technology evolved over the 20-year history of the NMDP [3][4][5][6]. Many of the intermediate-and highresolution typings of volunteer donors were performed because they were potential matches for a patient ("patientdirected typings").…”

Unrelated donor or partially matched related donor peripheral stem cell transplant with CD34+ selection and CD3+ addback for pediatric patients with leukemias

“…These methodologies have been previously tested and validated. 17,18 Conditioning and GVHD prophylaxis Conditioning included thiotepa 5 mg/kg days À6, À5, cyclophosphamide 60 mg/kg days À4, À3, and total body irradiation 1200 cGy/6 fractions days À2, À1, 0. Patients with CML in first chronic phase received anti-lymphocyte globulin 1.5 mg/kg days -2, -1, 0 for additional immunosuppression.…”