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Inventi Impact: Biomedical Analysis

Articles

  • Inventi:pba/51/12
    EVALUATION OF A SINGLE ROUND POLYMERASE CHAIN REACTION ASSAY USING DRIED BLOOD SPOTS FOR DIAGNOSIS OF HIV-1 INFECTION IN INFANTS IN AN AFRICAN SETTING
    30-Jun-2012 Research 2012 : July - September
    Bhavna H Chohan, Sandra Emery, Dalton Wamalwa, Grace John-Stewart, Maxwel Majiwa, Musa Ngayo,\r\nSteve Froggett, Julie Overbaugh

    Background: The aim of this study was to develop an economical ââ?¬Ë?in-houseââ?¬â?¢ single round polymerase chain\r\nreaction (PCR) assay using filter paper-dried blood spots (FP-DBS) for early infant HIV-1 diagnosis and to evaluate its\r\nperformance in an African setting.\r\nMethods: An ââ?¬Ë?in-houseââ?¬â?¢ single round PCR assay that targets conserved regions in the HIV-1 polymerase (pol) gene\r\nwas validated for use with FP-DBS; first we validated this assay using FP-DBS spiked with cell standards of known\r\nHIV-1 copy numbers. Next, we validated the assay by testing the archived FP-DBS (N = 115) from infants of known\r\nHIV-1 infection status. Subsequently this ââ?¬Ë?in-houseââ?¬â?¢ HIV-1 pol PCR FP-DBS assay was then established in Nairobi,\r\nKenya for further evaluation on freshly collected FP-DBS (N = 186) from infants, and compared with findings from\r\na reference laboratory using the Roche AmplicorÃ?® HIV-1 DNA Test, version 1.5 assay.\r\nResults: The HIV-1 pol PCR FP-DBS assay could detect one HIV-1 proviral copy in 38.7% of tests, 2 copies in 46.9%\r\nof tests, 5 copies in 72.5% of tests and 10 copies in 98.1% of tests performed with spiked samples. Using the\r\narchived FP-DBS samples from infants of known infection status, this assay was 92.8% sensitive and 98.3% specific\r\nfor HIV-1 infant diagnosis. Using 186 FP-DBS collected from infants recently defined as HIV-1 positive using the\r\ncommercially available Roche Amplicor v1.5 assay, 178 FP-DBS tested positive by this ââ?¬Ë?in-houseââ?¬â?¢ single-round HIV-1\r\npol PCR FP-DBS PCR assay. Upon subsequent retesting, the 8 infant FP-DBS samples that were discordant were\r\nconfirmed as HIV-1 negative by both assays using a second blood sample.\r\nConclusions: HIV-1 was detected with high sensitivity and specificity using both archived and more recently\r\ncollected samples. This suggests that this ââ?¬Ë?in-houseââ?¬â?¢ HIV-1 pol FP-DBS PCR assay can provide an alternative costeffective,\r\nreliable and rapid method for early detection of HIV-1 infection in infants.

    How to Cite this Article
    CC Compliant Citation: Chohan et al.: Evaluation of a single round polymerase chain reaction assay using dried blood spots for\r\ndiagnosis of HIV-1 infection in infants in an African setting. BMC Pediatrics 2011 11:18.
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