title: Design and evaluation of new, highly sensitive and specific primers for polymerase chain reaction detection of HIV-1 infected primary lymphocytes.
abstract: Primer pairs in the HIV-1 POL and ENV genes were evaluated by performing a PCR on lysed peripheral blood mononuclear cells (PBMCs) from 96 HIV-1 seropositive and 40 seronegative individuals originating from 16 different geographical localities in Africa, Europe and Haiti. A single PCR using primer pairs to the LTR, GAG and ENV regions and detection by radioactively labelled oligonucleotide probes was compared to a nested PCR scheme using newly designed POL and ENV primers which used ethidium-bromide staining of the amplified product on agarose gel. The newly designed POL nested primer pair was shown to be highly sensitive (93%) and specific (100%) for the detection of HIV-1 proviral DNA of very diverse geographical and genetic origin, including highly aberrant HIV-1 isolates. The sensitivity of the newly designed ENV primers was 68.7%, which does not differ significantly from the sensitivity of the classical primers, SK 68/69. Both ENV primers were unable to amplify two SIVcpz isolates from naturally infected chimpanzees.
authors.affiliation: Division of Microbiology, Institute of Tropical Medicine, Antwerp, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Fransen
authors.forename: P
authors.initials: P
authors.lastname: Zhong
authors.forename: H
authors.initials: H
authors.lastname: De Beenhouwer
authors.forename: G
authors.initials: G
authors.lastname: Carpels
authors.forename: M
authors.initials: M
authors.lastname: Peeters
authors.forename: J
authors.initials: J
authors.lastname: Louwagie
authors.forename: W
authors.initials: W
authors.lastname: Janssens
authors.forename: P
authors.initials: P
authors.lastname: Piot
authors.forename: G
authors.initials: G
authors.lastname: van der Groen
mesh_terms: D000349:Africa; D001483:Base Sequence; D017931:DNA Primers; D005060:Europe; D015752:Genes, env; D015751:Genes, pol; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D018023:HIV Seronegativity; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006205:Haiti; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008214:Lymphocytes / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014766:Viremia / Q000175:diagnosis* / Q000821:virology
chemical_list: D017931:DNA Primers