Performance Characteristics of Updated INNO-LiPA Assays for Molecular Typing of Human Leukocyte Antigen A (HLA-A), HLA-B, and HLA-DQB1 Alleles

Abstract

We carried out a multicenter performance evaluation of three new DNA-based human leukocyte antigen (HLA) typing assays: INNO-LiPA HLA-A Update, INNO-LiPA HLA-B Update, and INNO-LiPA HLA-DQB1 Update. After optimization, the accuracy rates were all 100%, and the final observed resolutions were 99.4, 92.4, and 85.6%, respectively. These rapid and easy-to-perform assays yielded results fully concordant with other DNA-based tissue typing tests.In order to optimize the selection of compatible organ and stem cell donors, DNA-based human leukocyte antigen (HLA) typing assays must not only be accurate but also take into account the continuing discovery of new HLA alleles (2). We recently updated three such HLA tests and then validated the new tests in two phases. First, we performed in-house assessments of the robustness of amplification and of the new probes by using well-characterized samples. In a second phase, reported below, we carried out external performance evaluations of the new assays.The three upgraded tests, INNO-LiPA HLA-A Update, INNO-LiPA HLA-B Update, and INNO-LiPA HLA-DQB1

HLA haplotypes and birth weight variation: is your future going to be light or heavy?

“…Each mother–child couple was analysed at low‐resolution level for HLA‐A , HLA‐B and HLA‐DRB1 polymorphisms, and children (cord blood donors) were analysed also at high resolution for HLA‐DRB1 polymorphisms. Low‐ and high‐resolution DNA analyses were carried out by polymerase chain reaction sequence‐specific primer method (Olerup, Saltsjöbaden, Sweden) and reverse PCR sequence‐specific oligonucleotide hybridization method (Innogenetics, Murex Biotech Limited, Ghent, Belgium) (35, 36).…”

Genetic epistasis between killer immunoglobulin-like receptors and human leukocyte antigens in Kawasaki disease susceptibility

“…We defined HLA-A and HLA-B polymorphisms at low-resolution level and HLA-C polymorphisms at high-resolution level by PCR with sequencespecific primers (INNO-TRAIN Diagnostik Gmbh, Kronberg, Germany) and/or by the reverse PCR sequence-specific oligonucleotide hybridization method (revPCR-SSO) (Innogenetics, Murex Biotech Limited, Ghent, Belgium), following the manufacturer's instruction. 34,35 Ambiguous typing results were resolved by direct sequencing. When only one allele was detected, the subject was considered homozygote.…”

Possible KIR-driven genetic pressure on the genesis and maintenance of specific HLA-A,B haplotypes as functional genetic blocks

“…Genomic DNA was genotyped by the PCR sequence-specific primer method (Olerup, Saltsjö baden, Sweden) and by the reverse PCR sequencespecific oligonucleotide hybridization method (Innogenetics, Murex Biotech Limited, Ghent, Belgium). 25,26 According to the international policy of FACT-Netcord and the Italian CB Banks, the CB units were typed for HLA-A and HLA-B polymorphisms at a low resolution level and for HLA-DRB1 at a high resolution before banking. According to the policy of IBMDR, the adult hematopoietic stem cell donors were typed for HLA-A and HLA-B and, more recently, for HLA-C polymorphisms at a low resolution level and for HLA-DRB1 at high resolution before data entry into the international database.…”