Allele typing of HLA-DQA1 by non-isotopic PCR-LIS-SSCP

ABBA MC; GÓMEZ MA; GOLIJOW CD

BRAZILIAN JOURNAL OF MEDICAL AND BIOLOGICAL RESEARCH

Associacâo Brasileira de Divulgacâ Científica

Lugar: Riberâo Preto, Brasil; Año: 2001 vol. 34 p. 867 - 869

0100-879X

In the present study we used a simple and reliable method for HLADQA1 allele typing based on the single-stranded conformation polymorphism (SSCP) properties of DNA molecules obtained by PCR. The technique consists of PCR amplification of a DNA fragment comprising the second exon of the HLA-DQA1 gene, amplicon denaturation using a low ionic strength solution (LIS), and electrophoresis on a small native polyacrylamide gel, followed by a rapid silver staining procedure. In order to validate the technique and to obtain the allele patterns for the DQA1 gene, 50 cervical samples were typed using this methodology and the commercial AmplitypeÆ HLA DQA1 Amplification and Typing kit. All the alleles detected with the kit were characterized by the LIS-SSCP approach. This procedure proved to be useful for population screening and typing of the DQA1 gene as well as for detecting new alleles or mutations in the donor-recipient molecular matching of HLA class II genes.