Comparison of serological and molecular (PCR-SSP) techniques of HLA-DR typing in clinical laboratory routine. 1997

G Woszczek, and M Borowiec, and M Miś, and M Górska, and M L Kowalski
Department of Clinical Immunology and Allergy, Medical University of Lódź, Poland. kowalsml@psk2.am.lodz.pl

Advances in molecular biology techniques allowed for introduction of PCR-based methods for HLA typing. In routine HLA typing for organ transplantation serological method is still being used as a standard, although molecular techniques are applied more and more often. The aim of our study was to compare HLA-DR typing using traditional serological method and PCR-SSP methodology in routine clinical laboratory. HLA-DR typing was performed using standard microcytotoxicity assay and PCR-SSP method in 28 patients referred to our Transplantation Immunology Unit for HLA typing. Comparison of results obtained by both methods revealed no discrepancies in 5 patients, in 12 patients the PCR-SSP typing showed additional DR antigens or splits of antigens. In 11 patients serological typing turned out to be impossible because of technical problems. Molecular PCR typing allowed for precise antigen determination in all the patients. Comparing both methods we found PCR-SSP HLA typing method very useful in routine HLA-DR determination, especially valuable in patients, in whom some problems in serological testing are expected.

UI MeSH Term Description Entries
D012150 Polymorphism, Restriction Fragment Length Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment. RFLP,Restriction Fragment Length Polymorphism,RFLPs,Restriction Fragment Length Polymorphisms
D006650 Histocompatibility Testing Identification of the major histocompatibility antigens of transplant DONORS and potential recipients, usually by serological tests. Donor and recipient pairs should be of identical ABO blood group, and in addition should be matched as closely as possible for HISTOCOMPATIBILITY ANTIGENS in order to minimize the likelihood of allograft rejection. (King, Dictionary of Genetics, 4th ed) Crossmatching, Tissue,HLA Typing,Tissue Typing,Crossmatchings, Tissue,HLA Typings,Histocompatibility Testings,Testing, Histocompatibility,Testings, Histocompatibility,Tissue Crossmatching,Tissue Crossmatchings,Tissue Typings,Typing, HLA,Typing, Tissue,Typings, HLA,Typings, Tissue
D006684 HLA-DR Antigens A subclass of HLA-D antigens that consist of alpha and beta chains. The inheritance of HLA-DR antigens differs from that of the HLA-DQ ANTIGENS and HLA-DP ANTIGENS. HLA-DR,Antigens, HLA-DR,HLA DR Antigens
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000483 Alleles Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product. Allelomorphs,Allele,Allelomorph
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain

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