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M9490068.TXT
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1994-09-03
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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).