Document 0068
 DOCN  M9490068
 TI    Detection of HIV-1 sequences in children using radioactive and
       colorimetric polymerase chain reactions.
 DT    9411
 AU    Giri AA; Lillo FB; McDermott JL; Jannuzzi C; Risso S; Fornia GL; Concedi
       DR; Varnier OE; Institute of Microbiology, School of Medicine, G.
       Gaslini; Children Hospital, Italy.
 SO    J Med Virol. 1994 Apr;42(4):414-9. Unique Identifier : AIDSLINE
       MED/94322025
 AB    The detection of HIV-1 proviral DNA in children born to seropositive
       mothers was studied using the polymerase chain reaction with either a
       radioactive electrophoretic method or a noval procedure that employs
       colorimetric microwell visualization. Peripheral blood mononuclear cell
       lysates from 18 HIV-1 infected children and 28 uninfected subjects were
       assayed for a 142 bp fragment of DNA from the gag region of HIV-1 using
       the primer pair SK145-431. Detection of amplified DNA was carried out by
       hybridization with a radiolabeled SK102 probe, or with a tagged SK102
       probe permitting colorimetric detection. The radioactive detection
       procedure demonstrated 100% specificity and correlated with the
       serological results. The assay was more sensitive than the p24 antigen
       test, but two false negative results were obtained. One was from a
       sample taken at 2 weeks, an age at which undetectable provirus levels
       were reported in almost all HIV-1 infected newborns. The second was
       probably due to a low copy number of proviral DNA, as positive results
       were obtained in all other (6) samples from this child. Comparative
       analysis in a limited number of specimens of radioactive and
       colorimetric detection following PCR revealed 100% specificity and
       comparable sensitivity with 4 discordant results. The results show that
       PCR is the best method for early diagnosis of HIV-1 infection in
       pediatric subjects. The study also demonstrated the value of a
       colorimetric detection method for PCR products. This colorimetric
       microwell plate procedure may prove a useful technique in routine
       diagnosis of HIV-1 infection in children.
 DE    Acquired Immunodeficiency Syndrome/*DIAGNOSIS  Child  Child, Preschool
       Colorimetry  DNA, Viral/*ANALYSIS  Female  Human  HIV Antibodies/BLOOD
       HIV-1/*GENETICS  Infant  Infant, Newborn  Male  *Polymerase Chain
       Reaction  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).