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Quantification of HIV-1 RNA in dried blood spots using real time quantitative PCR in resource limited settings

Emma Carine Iradukunda
Emma Carine Iradukunda
Chemistry Department
University of Georgia
Chemistry Building, Room 400
Analytical Seminar

Despite medical advancements to eradicate human immunodeficiency virus (HIV) around the world, HIV is still among the top leading causes of death in untreated newborn in resourcelimited countries. To address this issue, a confirmatory HIV infection test is required as soon as the baby is born to benefit from available HIV therapies such as antiretroviral therapy1.  However, the presence of maternal HIV antibodies transferred during pregnancy, childbirth as well as breastfeeding make the diagnosis very difficult to achieve, using standard antibody tests such as ELISA until the level of maternal antibody falls below limit of detection at 18 months.  It is then imperative to conduct a direct detection of the virus in infants below 18 months old.  Real-time polymerase chain reaction (rt-PCR) is one of the best analytical methods to detect HIV pro-viral DNA and RNA in newborns because of its capacity to detect HIV RNA viral load within few days after birth 2-3.

 To carry out this test, an adequate blood sample is needed. The use of dried blood spots (DBS) is preferred over whole blood samples in remote areas given that : 1) they are a minimally invasive and  its collection doesn’t require trained personnel;  2)  they can be collected at home and shipped to the testing facility using standard courier services under ambient conditions;  3) they present a very low biohazard risk compared to venipuncture method of blood collection and 4) all common analytic methods currently applied to traditional liquid samples can also be applied to DBS4-8. The overall aim of this seminar is to review the use of dried blood spots (DBS) in HIV surveillance using rt-PCR HIV-1 test in resource-limited areas. 

  1. Comeau AM, Pitt J, Hillyer GV, Landesman S, Bremer J, Chang BH, Lew J, Moye J, Grady GF, McIntosh K. J. Pediatr 1996, 129:111–118
  2. Arredondo M, Garrido C, Parkin N, Zahonero N, Bertagnolio S, Soriano V, de Mendoza C, J. Clin. Microbiol 2012, 50:569–572. 10.1128/JCM.00418-11
  3. Vidya M, Saravanan S, Rifkin S, Solomon SS, Waldrop G, Mayer KH, Solomon S, Balakrishnan P. J. Virol.  2012 181:177–181.
  4. Sherman GG, Stevens G, Jones SA, Horsfield P, Stevens WS. J. Acquir. Immune Defic. Syndr. 2005, 38:615–617. 10.1097
  5. Deep A,Kumar P, Kumar A, Thakkar A. Inten J Pharm sci 2012, 15: 90-4
  6. McDade TW. Am J Hum Biol 2014, 26:1-9.17.
  7. McDade TW, Williams S, Snodgrass JJ. Demography 2007, 44:899-925
  8. Hannon WH, Therrell BL. Hoboken (NJ): John Willey& Sons, Inc. 2014, P.1-5

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