Identification of HIV-1 infected infants and young children using real-time RT PCR and dried blood spots from Uganda and Cameroon

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...

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Bibliographic Details
Published in:Journal of virological methods 2007-09, Vol.144 (1), p.109-114
Main Authors: Ou, Chin-Yih, Yang, Hua, Balinandi, Steven, Sawadogo, Souleymane, Shanmugam, Vedapuri, Tih, Pius M., Adje-Toure, Christiane, Tancho, Sam, Ya, Leonard Kouadio, Bulterys, Marc, Downing, Robert, Nkengasong, John N.
Format: Article
Language:English
Subjects:
Cameroon
Child
DNA, Viral - blood
DNA, Viral - isolation & purification
Dried blood spots
Female
HIV Infections - diagnosis
HIV Infections - virology
HIV Seropositivity
HIV-1 - isolation & purification
Human immunodeficiency virus
Human immunodeficiency virus 1
Humans
Infant
Pediatric diagnosis
Real-time RT PCR
Reverse Transcriptase Polymerase Chain Reaction - methods
Sensitivity and Specificity
Uganda
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Summary: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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2007.04.003