Rapid and simple subtyping of the HLA-DRB3 gene in Graves' disease by using temperature-gradient gel electrophoresis.

Abstract	A HLA-DRB3-subtyping system that uses TGGE for analyzing DRB3 alleles was developed. The polymorphic second exon of the HLA-DRB3 gene was amplified from four homozygous typing cell lines, 223 healthy individuals, and 102 patients with Graves' disease by using the PCR. The PCR products were electrophoresed in a temperature gradient from 35 degrees C to 70 degrees C, and the resulting fragments were visualized by silver staining. Four DRB3 alleles (HLA-DRB30101, 0201, 0202, and 0301) were distinguished from one another by the migration of the corresponding homoduplex with the exception that DRB30201 was indistinguishable from DRB30202. The latter two alleles, however, were resolved by the artificial heteroduplexing approach. Arginine in position 74 of the DRB3 gene product (i.e., HLA-DRB30101) was significantly more frequent in Graves' patients than in controls. The relative risk conferred by the presence of the DRB30101 was 15.8 (p < or = 0.001). The presence of arginine in position 74 contributed to an etiologic fraction of 75% in our study population. The PCR-TGGE technique is a simple, nonisotopic method, which may be useful in rapid screening of large populations for HLA disease markers.
Authors	M Chen, W Maerz, B J Manfras, P Kuehnl, K H Usadel, B O Boehm
Journal	Human immunology (Hum Immunol) Vol. 36 Issue 3 Pg. 199-203 (Mar 1993) ISSN: 0198-8859 [Print] United States
PMID	8320138 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	DNA Probes, HLA DNA, Single-Stranded HLA-DR Antigens HLA-DRB3 Chains
Topics	Alleles Autoimmune Diseases (genetics, immunology) Base Sequence DNA Probes, HLA DNA, Single-Stranded Disease Susceptibility (immunology) Electrophoresis, Polyacrylamide Gel (methods) Genetic Predisposition to Disease Graves Disease (genetics, immunology) HLA-DR Antigens (genetics) HLA-DRB3 Chains Humans Molecular Sequence Data Nucleic Acid Conformation Polymerase Chain Reaction Risk Temperature

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.

Realize the full power of the drug-disease research graph!