Simultaneous Genotyping of Single Nucleotide Polymorphisms in the Il-1 Gene Complex by Multiplex Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

The interleukin-1 (IL-1) gene complex consists of the IL-1 alpha, IL-1 beta and IL-1 receptor antagonist genes. Single- nucleotide polymorphisms (SNP) in all three genes have been associated with human diseases. In this study, primers containing mismatches at 1-3 nucleotide positions were designed to incorporate a restriction site for endonuclease AlwNI or Xcml in the presence of allele-specific nucleotides at the polymorphic positions. Based on this technique, a simple and robust multiplex polymerase chain reaction/ restriction fragment length polymorphism ( multiplex PCR/RFLP ) assay was developed to determine simultaneously three to four informative SNPs (IL-1beta/ + 3954, IL-1beta-511 and IL -1Ra/9261 or IL-1alpha/-889, IL-1beta/-31, IL-1beta/5810 and IL-1Ra/11100 SNPs) in the IL-1 gene complex. (C) 2002 Elsevier Science B.V All rights reserved