title: LTR real-time PCR for HIV-1 DNA quantitation in blood cells for early diagnosis in infants born to seropositive mothers treated in HAART area (ANRS CO 01).
abstract: HIV-1 diagnosis in babies born to seropositive mothers is one of the challenges of HIV epidemics in children. A simple, rapid protocol was developed for quantifying HIV-1 DNA in whole blood samples and was used in the ANRS French pediatric cohort in conditions of prevention of mother-to-child transmission. A quantitative HIV-1 DNA protocol (LTR real-time PCR) requiring small blood volumes was developed. First, analytical reproducibility was evaluated on 172 samples. Results obtained on blood cell pellets and Ficoll-Hypaque separated mononuclear cells were compared in 48 adult HIV-1 samples. Second, the protocol was applied to HIV-1 diagnosis in infants in parallel with plasma HIV-RNA quantitation. This prospective study was performed in children born between May 2005 and April 2007 included in the ANRS cohort. The assay showed good reproducibility. The 95% detection cut-off value was 6 copies/PCR, that is, 40 copies/10(6) leukocytes. HIV-DNA levels in whole blood were highly correlated with those obtained after Ficoll-Hypaque separation (r = 0.900, P < 0.0001). A total of 3,002 specimens from 1,135 infants were tested. The specificity of HIV-DNA and HIV-RNA assays was 100%. HIV-1 infection was diagnosed in nine infants before age 60 days. HIV-DNA levels were low, underlining the need for sensitive assays when highly active antiretroviral therapy (HAART) has been given. The performances of this HIV-DNA assay showed that it is adapted to early diagnosis in children. The results were equivalent to those of HIV-RNA assay. HIV-DNA may be used even in masked primary infection in newborns whose mothers have received HAART.
authors.affiliation: Assistance Publique - Hôpitaux de Paris, CHU Necker-Enfants Malades, Service de Virologie, Paris, France. veronique.avettand@nck.aphp.fr
authors.forename: Véronique
authors.initials: V
authors.lastname: Avettand-Fènoël
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Chaix
authors.forename: Stéphane
authors.initials: S
authors.lastname: Blanche
authors.forename: Marianne
authors.initials: M
authors.lastname: Burgard
authors.forename: Corinne
authors.initials: C
authors.lastname: Floch
authors.forename: Kadidia
authors.initials: K
authors.lastname: Toure
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Allemon
authors.forename: Josiane
authors.initials: J
authors.lastname: Warszawski
authors.forename: Christine
authors.initials: C
authors.lastname: Rouzioux
mesh_terms: D000328:Adult; D019380:Anti-HIV Agents / Q000627:therapeutic use; D023241:Antiretroviral Therapy, Highly Active; D004279:DNA, Viral / Q000032:analysis / Q000097:blood* / Q000235:genetics; D005260:Female; D015658:HIV Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology / Q000635:transmission; D006679:HIV Seropositivity; D015497:HIV-1 / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007963:Leukocytes, Mononuclear / Q000821:virology; D009035:Mothers; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011446:Prospective Studies; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D019380:Anti-HIV Agents; D004279:DNA, Viral