ENZYME-LINKED DNA OLIGOTYPING - A PRACTICAL METHOD FOR CLINICAL HLA-DNA TYPING

The use of PCR and oligonucleotide hybridization has increased the acc uracy and resolution of typing for HLA class II alleles, but current p rocedures, performed in batches, take too long and are not suited for testing single samples. We have developed a typing method using enzyme -linked oligonucleotides and PCR products immobilized in 96-well trays . Trays preloaded with typing probes, covalently linked with alkaline phosphatase, have been kept for weeks at 4-degrees-C without loss of e nzyme-probe activity. Bound alkaline phosphatase was detected using a color reaction with enzymatic amplification which produces readings in 30 minutes. Coupled with a quick DNA preparation method, results can be obtained in about 4 hours. This method can be easily performed in s mall laboratories. It is accurate, reproducible, and sensitive, and wi ll make oligotyping for HLA alleles more convenient for testing clinic al samples.