title: Novel mutations and SNPs identified in CCR2 using a new comprehensive denaturing gradient gel electrophoresis assay.
abstract: A single nucleotide polymorphism (SNP) at codon 64 in the CC chemokine receptor 2 gene (CCR2 V64I) has been associated with a dominant effect of delaying disease progression from human immunodeficiency virus-1 (HIV-1) infection to acquired immunodeficiency syndrome (AIDS). The objective of our study was to design a comprehensive mutation detection assay for the entire coding region of the CCR2A and CCR2B gene transcripts, including all relevant splice site junctions and to identify novel mutations and SNPs within our predominantly African-based population, which could influence an individual's susceptibility to HIV-1 infection and/or progression to AIDS. The mutation detection assay, based on denaturing gradient gel electrophoresis (DGGE), allowed for the complete analysis of five individuals per denaturing gel. Our study cohort consisted of 102 HIV seropositive patients and 144 HIV seronegative controls from the diverse South African population. Application of the CCR2-DGGE assay resulted in the detection of two previously reported CCR2 polymorphisms, namely CCR2 V64I and CCR2 N260N, and 11 novel mutations, including seven SNPs occurring at high allelic frequencies within specific population groups of South Africa. The large number of novel mutations/SNPs identified, using the CCR2-DGGE assay, indicates the importance for comprehensive analysis of all candidate genes in host susceptibility to HIV-1 infection, specifically in the under-studied African-based populations.
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D015331:Cohort Studies; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005260:Female; D020022:Genetic Predisposition to Disease / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics; D015658:HIV Infections / Q000097:blood / Q000235:genetics; D018023:HIV Seronegativity / Q000235:genetics; D006679:HIV Seropositivity / Q000235:genetics; D015497:HIV-1 / Q000302:isolation & purification; D006801:Humans; D007532:Isoleucine / Q000235:genetics; D008297:Male; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000235:genetics*; D014633:Valine / Q000235:genetics
chemical_list: C515334:CCR2 protein, human; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D007532:Isoleucine; D014633:Valine