Application of random amplified polymorphic DNA PCR for genomic analysis of HIV-1-infected individuals

Mutat Res. 1998 Nov;406(1):25-31. doi: 10.1016/s1383-5726(98)00007-7.

Abstract

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) is a DNA fingerprinting technique used to detect genomic polymorphisms. We employed sixteen different RAPD-PCR 10-mer primers to amplify DNA from the peripheral blood mononuclear cells (PBMC) of 80 HIV-1-infected individuals. These individuals were previously identified as either heterozygotes (+ /delta32) and homozygotes (+/+) for the CCR5 locus by PCR with gene specific primers. Four of the sixteen randomly selected RAPD primers produced distinguishable banding profiles between CCR5 (+/delta32) heterozygotes and CCR5 (+/+ ) homozygotes. Direct sequencing of some RAPD-PCR products obtained with one of the four RAPD primers that were tested yielded clearly readable, but limited sequences, which were similar to portions of the previously published sequences for (+/+ ) homozygotes (98% similarity) and (+/delta32) heterozygotes (87% similarity) of the CCR5 alleles. Thus, the RAPD-PCR technique may be useful for the identification of human molecular markers that may correlate with susceptibility to HIV-1-infection, or differences in disease progression among HIV-l-infected individuals.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • DNA / blood
  • DNA Fingerprinting / methods
  • Genetic Carrier Screening
  • Genotype
  • HIV Infections / genetics*
  • HIV Infections / immunology
  • HIV-1*
  • Homozygote
  • Humans
  • Leukocytes, Mononuclear / immunology
  • Molecular Sequence Data
  • Polymorphism, Genetic*
  • Random Amplified Polymorphic DNA Technique*
  • Receptors, CCR5 / genetics*

Substances

  • Receptors, CCR5
  • DNA