RAPID DIRECT DETERMINATION OF HLA-DQBI-ASTERISK-0301 IN THE WHOLE-BLOOD OF NORMAL INDIVIDUALS AND CANCER-PATIENTS BY SPECIFIC POLYMERASE CHAIN-REACTION AMPLIFICATION

The HLA class II DQBl0301 allele is present at a higher frequency in patients with malignant melanoma than in Caucasian controls. Furthermo re, HLA-DQBl0301 identifies a group of melanoma patients presenting w ith relatively advanced disease, and independently identifies a group of melanoma patients more likely to have disease recurrence. A rapid s creening test for HLA-DQBl0301 may be useful in clinical research inv olving melanoma patients. Standard molecular oligotyping for HLA class II alleles using the polymerase chain reaction (PCR)-sequence specifi c oligonucleotide (SSO) method is relatively expensive, labor-intensiv e, and involves the use of radioisotope. We therefore developed an ine xpensive, rapid, non-radioactive method using sequence-specific primer s, peripheral whole blood as the substrate, and strictly defined react ion conditions in a single-step PCR to allow determination of the pres ence or absence of genomic HLA-DQBl0301. Comparison of the single-ste p PCR method with standard PCR-SSO oligotyping on 63 blinded samples f rom Caucasian melanoma patients demonstrated complete agreement betwee n the two methods in the detection of HLA-DQBl0301. Confirmatory test ing in 456 additional cancer patients and healthy controls showed a se nsitivity of 98.0% and a specificity of 99.4%. Single-step PCR is accu rate, rapid, inexpensive, and does not require radioisotope. These adv antages make it the procedure of choice for screening melanoma patient s and others for the presence of the HLA-DQBI 0301 allele.