Abstract
Objective: To develop tetra-primer PCR assays for detection of the CCR2-V64I, CCR5-A59029G and SDF1-G801A polymorphisms associated with HIV pathogenesis. Design and Methods: For each assay, two primers for the amplification of the gene locus are combined in one tube with two primers for the subsequent allele specific amplification (ASA). In the first set of cycles, pre-amplification of the gene region of interest is ensured by the gene specific primers. In the second set of cycles, lowering the annealing temperature allows ASA on the newly produced template. Results: Analysis of 90 DNA samples resulted in allele frequencies for CCR2-V64I, CCR5-A59029G and SDF1-G801A which are similar to other Caucasian cohorts. Furthermore, re-analysis of sequenced genomic DNA by tetra-primer PCR analysis (7–11 times) always showed identical results. Conclusion: Our set of single-tube assays allows rapid and reproducible genotyping of the CCR2-V64I, CCR5-A59029G and SDF1-G801A polymorphisms. These inexpensive but accurate assays are valuable for screening these polymorphisms in cohorts of HIV-infected patients.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
