A simple method of HLA-DRB typing using enzymatically amplified DNA and immobilized probes on microtiter plate.
We have developed a simple and economical method for HLA-DNA typing, called microtiter plate hybridization (PCR-MPH), which could replace standard PCR-SSO. This method is similar to that of an ELISA. Briefly, the PCR products labeled at the 5' termini with biotin were hybridized with probes immobilized on a microtiter well, and the bound PCR products were detected by streptavidin-conjugated enzymes followed by color development. A system for HLA-DRB1 "generic" typing (e.g., DR1, DR2), using microtiter wells coated with 12 different SSOs has been established. The HLA-DRB types classified using this method agreed well with those obtained by conventional serologic typing. The advantages of this microtiter plate-hybridization method for routine HLA-DNA typing are a short assay time, easy processing of large numbers of samples, and the potential for automation.
Institute for Biotechnological Research, Wakunaga Pharmaceutical, Hiroshima, Japan.
S
S
Kawai
S
S
Maekawajiri
K
K
Tokunaga
T
T
Juji
A
A
Yamane
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry; D005838:Genotype; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*
D004277:DNA, Single-Stranded; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D015345:Oligonucleotide Probes