title: Non-isotopic polymerase chain reaction methods for the detection of HIV-1 in Ugandan mothers and infants.
abstract: Two non-isotopic polymerase chain reaction (PCR) methods were evaluated by testing blood from 41 HIV-1-seropositive and 16 HIV-1-seronegative Ugandan mothers and 56 of their children (aged 0.5-15.0 months). Amplification of HIV-1 sequences was performed in duplicate using a biotinylated primer pair to the gag region (SK 462-431) and nested primer pairs (JA 17-20) to the pol region of HIV-1. gag sequences were hybridized using a microtiter plate coated with the SK 102 probe followed by colorimetric detection using an avidin-horseradish peroxidase conjugate and tetramethylbenzidine/peroxide substrate. pol sequences were detected on agarose gel stained with ethidium bromide. Results of HIV-1 PCR analysis showed that 40 out of 41 (98%) seropositive mothers and 10 out of 29 (34%) seropositive children had detectable HIV-1 gag and pol sequences. None of the 16 seronegative mothers nor 27 seronegative or Western blot-indeterminate children had detectable HIV-1 sequences. Our results suggest that non-isotopic PCR methods are sensitive, specific, and potentially useful in the early diagnosis of HIV-1 infection in developed and developing countries.
authors.affiliation: Department of Pathology, Case Western Reserve University, Cleveland, Ohio 44106.
authors.forename: J B
authors.initials: JB
authors.lastname: Jackson
authors.forename: C
authors.initials: C
authors.lastname: Ndugwa
authors.forename: F
authors.initials: F
authors.lastname: Mmiro
authors.forename: P
authors.initials: P
authors.lastname: Kataaha
authors.forename: L
authors.initials: L
authors.lastname: Guay
authors.forename: E A
authors.initials: EA
authors.lastname: Dragon
authors.forename: J
authors.initials: J
authors.lastname: Goldfarb
authors.forename: K
authors.initials: K
authors.lastname: Olness
mesh_terms: D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D015153:Blotting, Western; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015483:HIV Antibodies / Q000097:blood; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014454:Uganda
chemical_list: D004279:DNA, Viral; D015483:HIV Antibodies