Novel mutations identified using a comprehensive CCR5-denaturing gradient gel electrophoresis assay

Background: Most mutations detected for the gene for CC chemokine receptor 5 (CCR5) are either relatively specific to different population groups or r arely observed in Africans. Objectives: To develop a comprehensive mutation detection assay for the ent ire coding region of CCR5 and to identify novel mutations that may play a r ole in genetic susceptibility to HIV-1 infection, within the diverse South African population. Design: The study cohort consisted of 103 HIV-seropositive patients and 146 HIV-seronegative controls of predominantly African descent. Methods: A mut ation detection assay for the entire coding region of CCR5 was designed; th is included amplification of part of the coding region of CCR2. The assay w as based on denaturing gradient gel electrophoresis (DGGE) and allowed the complete analysis of samples from 10 individuals per denaturing gel. Results: The use of the CCR5-DCGE assay led to the identification of seven novel and six previously reported mutations. All novel mutations, including a common poly morphism at codon 35, occurred exclusively in non-Caucasians , indicating possible African origin. Conclusion: A comprehensive DGGE mutation detection assay has been develope d for the entire coding region of CCR5. Application of this assay resulted in the identification of novel CCR5 mutations, which may have a significant effect on the normal functioning of CCR5 and thus contribute to host varia bility and susceptibility to HIV-1 infection and/or progression to AIDS wit hin this population. (C) 2001 Lippincott Williams & Wilkins.