A new simplified method of gene typing.
SSP-PCR (sequence-specific primer) DNA typing was performed in Terasaki trays using 1.5 microliters of DNA, and the ethidium-stained PCR product was measured by direct fluorometric reading. Elimination of the gel electrophoresis step greatly simplified the SSP method. 17 serological DR specificities were discriminated for 239 DNA samples utilizing the new method, standard SSP, sequence-specific oligonucleotide probe (SSOP), and restriction fragment length polymorphism (PCR-RFLP). Results showed 98% concordance between the SSP-PCR assay and conventional methods. DRB1 alleles were determined by PCR-RFLP in 59 samples, by SSP in 110 samples, and by consensus (all methods) in the remaining samples.
Chia
D
D
UCLA Tissue Typing Laboratory, Department of Surgery, School of Medicine.
Terasaki
P
P
Chan
H
H
Acalinovich
A
A
Maruya
E
E
Saji
H
H
Ware
K
K
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics*; D006684:HLA-DR Antigens / Q000235:genetics; D059811:HLA-DRB1 Chains; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D017931:DNA Primers; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains