title: Identification of HIV-1 infected infants and young children using real-time RT PCR and dried blood spots from Uganda and Cameroon.
abstract: Serodiagnosis of HIV infection in infants born to HIV-infected mothers is problematic due to the prolonged presence of maternal antibodies in infants. Nucleic acid-based amplification assays have been used to overcome this problem. Here a simplified, one-tube, real-time, duplex reverse transcription PCR (RT PCR) assay is shown to detect HIV-1 total nucleic acid (TNA) isolated from dried blood spots. The detection of TNA, as opposed to DNA alone, increases the HIV target molecules and thus makes the assay more robust. This method was used to detect HIV from the DBS collected from HIV-1 exposed infants and young children in Uganda (n=128) and Cameroon (n=315). The gold-standards used were a plasma viral assay in Uganda and Amplicor DNA assay in Cameroon. The concordance of this real-time assay and the gold standards was 99.2% (127/128) and 99.4% (313/315) with the Ugandan and Cameroonian samples, respectively. This simple and cost-effective assay is potentially useful for the diagnosis of pediatric HIV infection and for evaluating programs to reduce mother-to-child transmission of HIV-1.
authors.affiliation: Division of HIV/AIDS Prevention, National Center of HIV, STD and TB Prevention, Global AIDS Program, Centers for Disease Control and Prevention, 1600 Clifton Road, Mail-stop A-12, Atlanta, GA 30333, USA. cho2@cdc.gov
authors.forename: Chin-Yih
authors.initials: CY
authors.lastname: Ou
authors.forename: Hua
authors.initials: H
authors.lastname: Yang
authors.forename: Steven
authors.initials: S
authors.lastname: Balinandi
authors.forename: Souleymane
authors.initials: S
authors.lastname: Sawadogo
authors.forename: Vedapuri
authors.initials: V
authors.lastname: Shanmugam
authors.forename: Pius M
authors.initials: PM
authors.lastname: Tih
authors.forename: Christiane
authors.initials: C
authors.lastname: Adje-Toure
authors.forename: Sam
authors.initials: S
authors.lastname: Tancho
authors.forename: Leonard Kouadio
authors.initials: LK
authors.lastname: Ya
authors.forename: Marc
authors.initials: M
authors.lastname: Bulterys
authors.forename: Robert
authors.initials: R
authors.lastname: Downing
authors.forename: John N
authors.initials: JN
authors.lastname: Nkengasong
mesh_terms: D002163:Cameroon; D002648:Child; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D006679:HIV Seropositivity; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014454:Uganda
chemical_list: D004279:DNA, Viral