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Typing of the granulocyte‐specific NA antigens by restriction fragment length polymorphism analysis
Author(s) -
Stein E.L.,
Bux J.,
Santoso S.,
MuellerEckhardt C.
Publication year - 1994
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1994.tb04939.x
Subject(s) - restriction enzyme , restriction fragment length polymorphism , microbiology and biotechnology , biology , polymerase chain reaction , primer (cosmetics) , genotype , typing , cleaved amplified polymorphic sequence , complementary dna , genetics , dna , gene , chemistry , organic chemistry
SUMMARY. 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 Fcgamma‐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.