Pro-Viral HIV-1 DNA PCR test for HIV Diagnosis in Infants.
Int Conf AIDS 2004 Jul 11-16; 15:(abstract no. B10283)
Enzama AR, Mugyenyi P, Kityo C Joint Clinical Research Centre, Kampala, Uganda
Background;The diagnosis of HIV-1 infection in neonates born to sero positive mothers presents two major dilemmas; the presence of maternal antibodies that preclude the use of serological tests, and the difficulty or impossibility of obtaining large volumes of blood for culture. Objectives;To evaluate a reliable method that uses small volumes of whole blood to diagnose HIV-1 in infants, as a surrogate serological test. To monitor the rate of Pediatric sero-conversion due to breastfeeding. Method;Between July 2001 and January 2003 whole blood samples drawn from babies at birth, week 4, week 6, month 3 and month 6 were tested at the Joint clinical Research Centre Laboratories. Two aliquots of cell pellet were prepared from 200ul of blood from each draw and plasma separated for storage. Qualitative HIV-1 DNA PCR was performed on one pellet at each visit to determine the presence of HIV-1 Pro-viral DNA. Where there was a positive DNA test the second pellet was assayed in duplicate and if still positive RNA PCR was done on the stored plasma to determine the viral load. If the viral load was above 400 copies/ml, HIV-1 ELISA was performed on the respective plasma to determine the presence of antibodies to HIV due to active immunity. ResultA negative DNA test at birth meant the baby was not HIV infected. A Positive DNA test with negative ELISA and Viral load<400 copies/ml indicated HIV infection. Subsequent DNA tests were used to determine sero-conversion. Conclusion;HIV-1 DNA test is a reliable method for infant and window stage diagnosis of HIV infection. HIV negative babies at birth can be isolated to prevent infection through breast milk. This study is an eye opener to determining the most risky period of pediatric infection during the 6 months of breast feeding.
Keywords: AEGIS, Polymerase Chain Reaction, HIV-1, HIV Infections, Viral Load, DNA, HIV Antibodies, HIV Seropositivity, DNA, Viral, HIV, HIV Core Protein p24, Hematologic Tests, Branched DNA Signal Amplification Assay, Laboratory Techniques and Procedures, DNA Primers, CD4 Lymphocyte Count, Anti-HIV Agents, Acquired Immunodeficiency Syndrome, Breast Feeding, Infant, Humans, Child, immunology, diagnosis
