BACKGROUND: The genotype of human leukocyte antigen (HLA) alleles has important effect on organ transplantation and medicolegal expertise. The presented genotyping can not reach high-flux or integration, without accuracy or repetitiveness.
OBJECTIVE: To evaluate the accuracy and reliability of self-fabricated oligoneucleotide array by comparing with polymerase chain reaction with sequence-specific primers (PCR-SSP) in identification of HLA-DQA1 alleles.
METHODS: A total of 100 clinical samples were enrolled in the study. HLA-DQA1 genotyping were performed by PCR-SSP and oligoneucleotide array, respectively. A pairs of group-specific primers were designed according to the sequence polymorphism of HLA-DQA1 exon two. The target DNA was asymmetrically amplified with the labeled sense primer. The labeled PCR products were hybridized with the specific allele typing probes immobilized on a glass support, and the signals were scanned by scanner and then analyzed. The discordant samples by arrays and PCR-SSP were verified by sequencing.
RESULTS AND CONCLUSION: All 100 samples have been genotyped by oligoneucleotide array and PCR-SSP successfully. The coincidence rate was 94%. Four homozygous samples typed by PCR-SSP were actually heterozygous by array. The other two unidentified samples were typed by sequencing. The results showed that a mistake for one sample was made by PCR-SSP or array. In reproducible tests, the signal reappear rate was 95%. That is, the self-fabricated oligoneucleotide array with ideal stability, specificity and sensitivity, which provide a suitable plateau for HLA-DQA1 genotyping domestically.