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DETERMINATION OF DQB1 ALLELES USING PCR AMPLIFICATION AND ALLELE‐SPECIFIC PRIMERS
Author(s) -
Lepage V.,
Ivanova R.,
Loste M.N.,
Mallet C.,
Douay C.,
Naoumova E.,
Charron D.
Publication year - 1995
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.1995.tb00256.x
Subject(s) - genotyping , polymerase chain reaction , typing , allele , biology , restriction fragment length polymorphism , variants of pcr , genetics , microbiology and biotechnology , genotype , gene
SUMMARY Molecular genotyping of HLA class II genes is commonly carried out using polymerase chain reaction (PCR) in combination with sequence‐specific oligotyping (PCR‐SSO) or a combination of the PCR and restriction fragment length polymorphism methods (PCR‐RFLP). However, the identification of the DQB1 type by PCR‐SSO and PCR‐RFLP is very time‐consuming which is disadvantageous for the typing of cadaveric organ donors. We have developed a DQB1 typing method using PCR in combination with allele‐specific amplification (PCR‐ASA), which allows the identification of the 17 most frequent alleles in one step using seven amplification mixtures. PCR allele‐specific amplification HLA‐DQB1 typing is easy to perform, and the results are easy to interpret in routine clinical practice. The PCR‐ASA method is therefore better suited to DQB1 typing for organ transplantation than other methods.