Smarter Decisions,
Better Care

UpToDate synthesizes the most recent medical information into evidence-based practical recommendations clinicians trust to make the right point-of-care decisions.

  • Rigorous editorial process: Evidence-based treatment recommendations
  • World-Renowned physician authors: over 5,100 physician authors and editors around the globe
  • Innovative technology: integrates into the workflow; access from EMRs

Choose from the list below to learn more about subscriptions for a:


Subscribers log in here


Techniques and interpretation of HIV-1 RNA quantitation

INTRODUCTION

Human immunodeficiency virus type 1 (HIV-1) RNA can be measured using qualitative or quantitative techniques. Qualitative testing (commonly referred to as nucleic acid testing or NAT) is used as a screening test to identify HIV infected individuals, such as screening possible blood donors. Quantification of HIV-RNA (viral load measurements) can be used as a diagnostic test in certain situations; however, the HIV viral load is primarily used for management/monitoring of HIV-1 infected individuals.

This topic will address the laboratory methods for quantitation of HIV-1 RNA and the use of viral load for clinical management. Information on nucleic acid, HIV-2 RNA, and CD-4 cell count testing is found elsewhere. (See "Blood donor screening: Laboratory testing", section on 'HIV-1 and HIV-2' and "Clinical manifestations and diagnosis of HIV-2 infection", section on 'Testing for HIV-2 infection' and "Techniques and interpretation of measurement of the CD4 cell count in HIV-infected patients".)

IMPORTANCE OF HIV VIRAL LOAD MEASUREMENTS

Studies have shown HIV-1 RNA levels to be a predictor of the time to progression to acquired immunodeficiency syndrome (AIDS) and death that is independent of CD4 cell counts [1-6]. Viral load measurements are also useful in determining when to initiate antiretroviral therapy, and in monitoring the response to such therapy [7-11]. (See "Selecting antiretroviral regimens for the treatment-naïve HIV-infected patient" and "Patient monitoring during HIV antiretroviral therapy".)

In specific situations (neonatal infection and acute infection), HIV-1 RNA levels also may be useful in establishing the diagnosis of HIV infection, but HIV antibody tests are primarily used for this purpose.

LABORATORY METHODS FOR QUANTITATION OF HIV-1 RNA

There are four commercial assays that have been approved by the United States Food and Drug Administration (FDA) to quantify HIV-1 RNA from plasma samples:

           

Subscribers log in here

To continue reading this article, you must log in with your personal, hospital, or group practice subscription. For more information or to purchase a personal subscription, click below on the option that best describes you:
Literature review current through: Nov 2014. | This topic last updated: Jan 30, 2014.
The content on the UpToDate website is not intended nor recommended as a substitute for medical advice, diagnosis, or treatment. Always seek the advice of your own physician or other qualified health care professional regarding any medical questions or conditions. The use of this website is governed by the UpToDate Terms of Use ©2014 UpToDate, Inc.
References
Top
  1. Mellors JW, Rinaldo CR Jr, Gupta P, et al. Prognosis in HIV-1 infection predicted by the quantity of virus in plasma. Science 1996; 272:1167.
  2. O'Brien TR, Blattner WA, Waters D, et al. Serum HIV-1 RNA levels and time to development of AIDS in the Multicenter Hemophilia Cohort Study. JAMA 1996; 276:105.
  3. O'Brien WA, Hartigan PM, Martin D, et al. Changes in plasma HIV-1 RNA and CD4+ lymphocyte counts and the risk of progression to AIDS. Veterans Affairs Cooperative Study Group on AIDS. N Engl J Med 1996; 334:426.
  4. Katzenstein DA, Hammer SM, Hughes MD, et al. The relation of virologic and immunologic markers to clinical outcomes after nucleoside therapy in HIV-infected adults with 200 to 500 CD4 cells per cubic millimeter. AIDS Clinical Trials Group Study 175 Virology Study Team. N Engl J Med 1996; 335:1091.
  5. Shearer WT, Quinn TC, LaRussa P, et al. Viral load and disease progression in infants infected with human immunodeficiency virus type 1. Women and Infants Transmission Study Group. N Engl J Med 1997; 336:1337.
  6. Mellors JW, Muñoz A, Giorgi JV, et al. Plasma viral load and CD4+ lymphocytes as prognostic markers of HIV-1 infection. Ann Intern Med 1997; 126:946.
  7. Saag MS, Holodniy M, Kuritzkes DR, et al. HIV viral load markers in clinical practice. Nat Med 1996; 2:625.
  8. Hughes MD, Johnson VA, Hirsch MS, et al. Monitoring plasma HIV-1 RNA levels in addition to CD4+ lymphocyte count improves assessment of antiretroviral therapeutic response. ACTG 241 Protocol Virology Substudy Team. Ann Intern Med 1997; 126:929.
  9. O'Brien WA, Hartigan PM, Daar ES, et al. Changes in plasma HIV RNA levels and CD4+ lymphocyte counts predict both response to antiretroviral therapy and therapeutic failure. VA Cooperative Study Group on AIDS. Ann Intern Med 1997; 126:939.
  10. Yeni PG, Hammer SM, Hirsch MS, et al. Treatment for adult HIV infection: 2004 recommendations of the International AIDS Society-USA Panel. JAMA 2004; 292:251.
  11. Panel on Antiretroviral Guidelines for Adults and Adolescents. Guidelines for the use of antiretroviral agents in HIV-1-infected adults and adolescents. Department of Health and Human Services. Available at http://www.aidsinfo.nih.gov/contentfiles/lvguidelines/adultandadolescentgl.pdf (Accessed on October 31, 2013).
  12. Mulder J, McKinney N, Christopherson C, et al. Rapid and simple PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma: application to acute retroviral infection. J Clin Microbiol 1994; 32:292.
  13. Pachl C, Todd JA, Kern DG, et al. Rapid and precise quantification of HIV-1 RNA in plasma using a branched DNA signal amplification assay. J Acquir Immune Defic Syndr Hum Retrovirol 1995; 8:446.
  14. Schumacher W, Frick E, Kauselmann M, et al. Fully automated quantification of human immunodeficiency virus (HIV) type 1 RNA in human plasma by the COBAS AmpliPrep/COBAS TaqMan system. J Clin Virol 2007; 38:304.
  15. Gueudin M, Plantier JC, Lemée V, et al. Evaluation of the Roche Cobas TaqMan and Abbott RealTime extraction-quantification systems for HIV-1 subtypes. J Acquir Immune Defic Syndr 2007; 44:500.
  16. Foulongne V, Montes B, Didelot-Rousseau MN, Segondy M. Comparison of the LCx human immunodeficiency virus (HIV) RNA quantitative, RealTime HIV, and COBAS AmpliPrep-COBAS TaqMan assays for quantitation of HIV type 1 RNA in plasma. J Clin Microbiol 2006; 44:2963.
  17. Berger A, Scherzed L, Stürmer M, et al. Comparative evaluation of the Cobas Amplicor HIV-1 Monitor Ultrasensitive Test, the new Cobas AmpliPrep/Cobas Amplicor HIV-1 Monitor Ultrasensitive Test and the Versant HIV RNA 3.0 assays for quantitation of HIV-1 RNA in plasma samples. J Clin Virol 2005; 33:43.
  18. Damond F, Roquebert B, Bénard A, et al. Human immunodeficiency virus type 1 (HIV-1) plasma load discrepancies between the Roche COBAS AMPLICOR HIV-1 MONITOR Version 1.5 and the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 assays. J Clin Microbiol 2007; 45:3436.
  19. Cao Y, Ho DD, Todd J, et al. Clinical evaluation of branched DNA signal amplification for quantifying HIV type 1 in human plasma. AIDS Res Hum Retroviruses 1995; 11:353.
  20. Lin HJ, Myers LE, Yen-Lieberman B, et al. Multicenter evaluation of quantification methods for plasma human immunodeficiency virus type 1 RNA. J Infect Dis 1994; 170:553.
  21. Elbeik T, Charlebois E, Nassos P, et al. Quantitative and cost comparison of ultrasensitive human immunodeficiency virus type 1 RNA viral load assays: Bayer bDNA quantiplex versions 3.0 and 2.0 and Roche PCR Amplicor monitor version 1.5. J Clin Microbiol 2000; 38:1113.
  22. Farzadegan H, Hoover DR, Astemborski J, et al. Sex differences in HIV-1 viral load and progression to AIDS. Lancet 1998; 352:1510.
  23. Jain V, Liegler T, Kabami J, et al. Assessment of population-based HIV RNA levels in a rural east African setting using a fingerprick-based blood collection method. Clin Infect Dis 2013; 56:598.
  24. Dickover RE, Herman SA, Saddiq K, et al. Optimization of specimen-handling procedures for accurate quantitation of levels of human immunodeficiency virus RNA in plasma by reverse transcriptase PCR. J Clin Microbiol 1998; 36:1070.
  25. Holodniy M, Mole L, Yen-Lieberman B, et al. Comparative stabilities of quantitative human immunodeficiency virus RNA in plasma from samples collected in VACUTAINER CPT, VACUTAINER PPT, and standard VACUTAINER tubes. J Clin Microbiol 1995; 33:1562.
  26. Holodniy M, Rainen L, Herman S, Yen-Lieberman B. Stability of plasma human immunodeficiency virus load in VACUTAINER PPT plasma preparation tubes during overnight shipment. J Clin Microbiol 2000; 38:323.
  27. García-Bujalance S, Ladrón de Guevara C, González-García J, et al. Elevation of viral load by PCR and use of plasma preparation tubes for quantification of human immunodeficiency virus type 1. J Microbiol Methods 2007; 69:384.
  28. Griffith BP, Mayo DR. Increased levels of HIV RNA detected in samples with viral loads close to the detection limit collected in Plasma Preparation Tubes (PPT). J Clin Virol 2006; 35:197.
  29. Erice A, Brambilla D, Bremer J, et al. Performance characteristics of the QUANTIPLEX HIV-1 RNA 3.0 assay for detection and quantitation of human immunodeficiency virus type 1 RNA in plasma. J Clin Microbiol 2000; 38:2837.
  30. Sun R, Ku J, Jayakar H, et al. Ultrasensitive reverse transcription-PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma. J Clin Microbiol 1998; 36:2964.
  31. Holodniy M, Mole L, Winters M, Merigan TC. Diurnal and short-term stability of HIV virus load as measured by gene amplification. J Acquir Immune Defic Syndr 1994; 7:363.
  32. Brambilla D, Reichelderfer PS, Bremer JW, et al. The contribution of assay variation and biological variation to the total variability of plasma HIV-1 RNA measurements. The Women Infant Transmission Study Clinics. Virology Quality Assurance Program. AIDS 1999; 13:2269.
  33. Donovan RM, Bush CE, Markowitz NP, et al. Changes in virus load markers during AIDS-associated opportunistic diseases in human immunodeficiency virus-infected persons. J Infect Dis 1996; 174:401.
  34. Mole L, Ripich S, Margolis D, Holodniy M. The impact of active herpes simplex virus infection on human immunodeficiency virus load. J Infect Dis 1997; 176:766.
  35. O'Brien WA, Grovit-Ferbas K, Namazi A, et al. Human immunodeficiency virus-type 1 replication can be increased in peripheral blood of seropositive patients after influenza vaccination. Blood 1995; 86:1082.
  36. Staprans SI, Hamilton BL, Follansbee SE, et al. Activation of virus replication after vaccination of HIV-1-infected individuals. J Exp Med 1995; 182:1727.
  37. Hu DJ, Dondero TJ, Rayfield MA, et al. The emerging genetic diversity of HIV. The importance of global surveillance for diagnostics, research, and prevention. JAMA 1996; 275:210.
  38. Cartwright CP. The changing epidemiology of HIV/AIDS at a Minnesota hospital: impact of demographic change and viral diversity. J Med Virol 2006; 78 Suppl 1:S19.
  39. Lin HH, Gaschen BK, Collie M, et al. Genetic characterization of diverse HIV-1 strains in an immigrant population living in New York City. J Acquir Immune Defic Syndr 2006; 41:399.
  40. Parekh B, Phillips S, Granade TC, et al. Impact of HIV type 1 subtype variation on viral RNA quantitation. AIDS Res Hum Retroviruses 1999; 15:133.
  41. Jagodzinski LL, Wiggins DL, McManis JL, et al. Use of calibrated viral load standards for group M subtypes of human immunodeficiency virus type 1 to assess the performance of viral RNA quantitation tests. J Clin Microbiol 2000; 38:1247.
  42. Swanson P, Huang S, Abravaya K, et al. Evaluation of performance across the dynamic range of the Abbott RealTime HIV-1 assay as compared to VERSANT HIV-1 RNA 3.0 and AMPLICOR HIV-1 MONITOR v1.5 using serial dilutions of 39 group M and O viruses. J Virol Methods 2007; 141:49.
  43. Rouet F, Chaix ML, Nerrienet E, et al. Impact of HIV-1 genetic diversity on plasma HIV-1 RNA Quantification: usefulness of the Agence Nationale de Recherches sur le SIDA second-generation long terminal repeat-based real-time reverse transcriptase polymerase chain reaction test. J Acquir Immune Defic Syndr 2007; 45:380.