Typing of the granulocyte-specific NA antigens by restriction fragment length polymorphism analysis.
An RNA-based method has been developed to genotype donors for the granulocyte-specific alloantigens NA1 and NA2. mRNA was isolated from granulocytes, reversely transcribed into cDNA and amplified using an Fc-gamma-receptor III-1 sequence-specific primer in the polymerase chain reaction (PCR). PCR products were analysed by restriction fragment length polymorphism (RFLP) using the restriction endonuclease Taq I, which provided a distinct restriction fragment pattern corresponding to the NA alleles. 17 donors were typed by PCR-RFLP and the results were in close accordance with those obtained by serological phenotyping by granulocyte immunofluorescence and the antigen capture assay MAIGA.
Stein
E L
EL
Institute for Clinical Immunology and Transfusion Medicine, University of Giessen, Germany.
Bux
J
J
Santoso
S
S
Mueller-Eckhardt
C
C
D000954:Antigens, Surface / Q000097:blood*; D001483:Base Sequence; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D007118:Immunoassay; D007519:Isoantigens / Q000097:blood*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017452:Receptors, IgG / Q000235:genetics
D000954:Antigens, Surface; D018076:DNA, Complementary; D007519:Isoantigens; D017452:Receptors, IgG