Información del artículo
Resumen
Las técnicas de cuantificación viral tienen cada vez más demanda en la microbiología clínica, bien para la evolución de pacientes trasplantados, bien para el seguimiento a largo plazo de enfermedades crónicas como los virus de la inmunodeficiencia humana (VIH) y de la hepatitis C (VHC). Las diferentes compañías farmacéuticas, interesadas en el desarrollo de metodologías eficientes y precisas para el diagnóstico y la correcta cuantificación viral han convergido en los últimos 2 años en la técnica de reacción en cadena de la polimerasa (PCR) en tiempo real aplicada a la cuantificación del VIH y del VHC, para incrementar la sensibilidad, la precisión, la linealidad y la correcta detección de la diversidad genómica de estos dos virus respecto a las técnicas clásicas que se venían aplicando desde finales de la década de 1990. La PCR en tiempo real es la gran herramienta del futuro inmediato, por su sensibilidad y precisión; sin embargo, se debe dar respuesta a nuevas preguntas, como si la constante variabilidad genética en ambos virus puede afectar progresivamente a la correcta cuantificación viral, situación difícilmente detectable al existir una uniformidad metodológica.
Palabras clave:
Virus de la inmunodeficiencia humana tipo 1
Hepatitis C
Carga viral
Abstract
Viral load techniques are more and more demanded in Clinical Microbiology, regarding with transplanted patients or long time follow-up of chronic diseases as those caused by human inmunodeficiency (HIV) and hepatitis C (HCV) viruses. In the last 2 years, pharmaceutical companies, interested to develop more efficient and accurate methods for the diagnosis and correct viral quantification of HIV and HCV, have converged in the real time-polymerase chain reaction (PCR) technique. This process is due to the increased sensitiviy, accuracy, linearity and correct detection of genomic viral variants of real time PCR techniques, in comparison with classical molecular methods applied since the nineties of the past century. In spite real time PCR appears as the best tool for the immediate future, new questions regarding the high variability of these viruses should be considered. This could affect the correctness of viral quantifications, while being difficult to detect it because of the methodological uniformity in the clinical laboratories.
Keywords:
Human immunodeficiency virus type 1
Hepatitis C
Viral load
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Bibliografía
[1.]
J.L. Fahey, J.M. Taylor, R. Detels, B. Hofmann, R. Melmed, P. Nishanian, et al.
N Engl J Med, 322 (1990), pp. 166-172
[2.]
J.W. Mellors, C.R. Rinaldo, P. Gupta, R.M. White, J.A. Todd, L.A. Kingsley.
Prognosis in HIV-1 infection predicted by the quantity of virus in plasma.
Science, 272 (1996), pp. 1167-1170
[3.]
J.W. Mellors, L.A. Kingsley, C.R. Rinaldo, J.A. Todd, B.S. Hoo, R.P. Kokka, et al.
Quantitation of HIV-1 RNA in plasma predicts outcome after seroconversion.
Ann Intern Med, 122 (1995), pp. 573-579
[4.]
F. de Wolf, I. Spijkerman, P. Schellekens, M. Langendam, C. Kuiken, M. Bakker, et al.
AIDS prognosis based on HIV-1 RNA, CD4+ T-cell count and function: markers with reciprocal predictive value over time after seroconversion.
AIDS, 11 (1997), pp. 1799-1806
[5.]
J.W. Mellors, J.B. Margolick, J.P. Phair, C.R. Rinaldo, R. Detels, L.P. Jacobson, et al.
Prognostic value of HIV-1 RNA, CD4 cell count, and CD4 cell count slope for progression to AIDS and death in untreated HIV-1 infection.
JAMA, 297 (2007), pp. 2349-2350
[6.]
S.E. Langford, J. Ananworanich, D.A. Cooper.
Predictors of disease progression in HIV infection: a review.
AIDS Res Ther, 4 (2007), pp. 11
[7.]
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.; 1-139 [actualizado: 3 de noviembre de 2008]. Disponible en: http://www.aidsinfo.nih.gov/ContentFiles/AdultandAdolescentGL.pdf.
[8.]
Anónimo, WHO. Recommendations for antiretroviral therapy for HIV infection in adults and adolescents. Key findings and recommendations from consultations with people living with HIV [actualizado: 31 de agosto de 2009]. Disponible en:http://www.gnpplus.net/images/stories/20090831_findings_and_recommendations_plhiv_consultations.pdf.
[9.]
Panel de expertos de GESIDA y Plan Nacional sobre el SIDA. Recomendaciones de GESIDA/Plan Nacional sobre el SIDA respecto al tratamiento antirretroviral en adultos infectados por el virus de la inmunodeficiencia humana [actualizado: febrero de 2009]. Disponible en: http://www.msps.es/ciudadanos/enfLesiones/enfTransmisibles/sida/docs/recomendacionesGesidaPNSTARVfebrero2009.pdf.
[10.]
K. Mullis, F. Faloona, S. Scharf, R. Saiki, G. Horn, H. Erlich.
Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction?.
Cold Spring Harbor Symposium on Quantitative Biology, 51 (1986), pp. 263
[11.]
FDA Advisor Committee Recommends Approval for Roche's Amplicor HIV Monitor Test. The first standardized PCR test to measure HIV levels in blood. 1996. Disponible en: http://www.aegis.com/news/pr/1996/PR960326.html.
[12.]
N. Zanchetta, G. Nardi, L. Tocalli, L. Drago, C. Bossi, F.R. Pulvirenti, et al.
Evaluation of the abbott LCx HIV-1 RNA quantitative, a new assay for quantitative determination of human immunodeficiency virus type 1 RNA.
J Clin Microbiol, 38 (2000), pp. 3882-3886
[13.]
Abbott LCx probes system. Abbott Laboratories, folleto técnico.
[14.]
T. Kievits, B. van Gemen, D. van Strijp, R. Schukkink, M. Dircks, H. Adriaanse, et al.
NASBA isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection.
J Virol Methods, 35 (1991), pp. 273-286
[15.]
NucliSens HIV-1 QT BioMérieux, package insert 2001. Disponible en: http://www.fda.gov/downloads/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/PremarketApprovalsPMAs/ucm091370.pdf.
[16.]
M.S. Urdea, J.C. Wilber, T. Yeghiazarian, J.A. Todd, D.G. Kern, S.J. Fong, et al.
Direct and quantitative detection of HIV-1 RNA in human plasma with a branched DNA signal amplification assay.
AIDS, 7 (1993), pp. S11-S14
[17.]
Siemens Versant HIV-1 RNA 3.0 Assay Siemens Mediacal Solution 2002. Disponible en: http://www.fda.gov/downloads/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/PremarketApprovalsPMAs/ucm091276.pdf.
[18.]
Y. Schmitt.
Performance characteristics of quantification assays for human immunodeficiency virus type 1 RNA.
J Clin Virol, 20 (2001), pp. 31-33
[19.]
D.N. Chernoff.
The significance of HIV viral load assay precision: a review of the package insert specifications of two commercial kits.
J Int Assoc Physicians AIDS Care, 1 (2002), pp. 134-140
[20.]
F. Rouet, M.L. Chaix, E. Nerrienet, N. Ngo-Giang-Huong, J.C. Plantier, Burgard, 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 Inmune Defic Syndr, 45 (2007), pp. 380-388
[21.]
J.F. Drexler, L.K. de Souza Luna, C. Pedroso, D.B. Pedral-Sampaio, A.T.L. Queiroz, E. Brites, et al.
Rates of and reasons for failure of commercial human immunodeficiency virus type 1 viral load assays in Brazil.
J Clin Microbiol, 45 (2007), pp. 2061-2063
[22.]
J.M. Raboud, S. Rae, R.S. Hogg, B. Yip, C.H. Sherlock, P.R. Harrigan, et al.
Suppression of plasma virus load below the detection limit of a human immunodeficiency virus kit is associated with longer virologic response than suppression below the limit of quantitation.
J Infect Dis, 180 (1999), pp. 1347-1350
[23.]
T.L. Kieffer, M.M. Finucane, R.E. Nettles, T.C. Quinn, K.W. Broman, S.C. Ray, et al.
Genotypic analysis of HIV-1 drug resistance at the limit of detection: virus production without evolution in treated adults with undetectable HIV loads.
J Infect Dis, 189 (2004), pp. 1452-1465
[24.]
R.E. Nettles, T.L. Kieffer, R.P. Simmons, J. Cofrancesco, R.D. Moore, J.E. Gallant, et al.
Genotypic resistance in HIV-1-infected patients with persistently detectable lowlevel viremia while receiving highly active antiretroviral therapy.
Clin Infect Dis, 39 (2004), pp. 1030-1037
[25.]
A. Holguin, M. de Mulder, G. Yebra, M. Lopez, V. Soriano.
Increase of non-B subtypes and recombinants among newly diagnosed HIV-1 native Spaniards and immigrants in Spain.
Curr HIV Res, 6 (2008), pp. 327-334
[26.]
A.M. Geretti.
HIV-1 subtypes: epidemiology and significance for HIV management.
Curr Opin Infect Dis, 19 (2006), pp. 1-7
[27.]
M.L. Wong, J.F. Medrano.
Real-time PCR for mRNA quantification.
BioTechiques, 39 (2005), pp. 75-85
[28.]
Roche Molecular System COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, 48 Tests. 2007. Disponible en: http://www.fda.gov/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/PremarketApprovalsPMAs/ucm091618.htm.
[29.]
Abbott Molecular. Abbott RealTime HIV-1 Amplification Reagent Kit, Abbott. 2007. Disponible en: http://www.fda.gov/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/PremarketApprovalsPMAs/ucm089797.htm.
[30.]
Siemens Healthcare Diagnostics. Versant HIV-1 RNA 1.0 Assay (kPCR). 2008. Folleto técnico.
[31.]
P. van Deursen, A. Verhoeven, P. de Bie, L. Bertens, J. de Jong.
Measuring human immunodeficiency virus type 1 RNA loads in dried blood spot specimens using NucliSENS EasyQ HIV-1 v2.0.
ECCMID, (2009),
[32.]
J.Y. Choi, E.J. Kim, H.J. Rho, J.Y. Kim, O.K. Kwon, J.H. Lee, et al.
Evaluation of the NucliSens EasyQ HIV-1 v1.1 and RealTime HIV-1 kits for quantitation of HIV-1 RNA in plasma.
J Virol Methods, 161 (2009), pp. 7-11
[33.]
V. Foulongne, B. Montes, M.N. Didelot-Rousseau, M. Segondy.
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, 44 (2006), pp. 2963-2966
[34.]
H. Gatanaga, K. Tsukada, H. Honda, J. Tanuma, H. Yazaki, T. Watanabe, et al.
Detection of HIV type 1 load by the Roche Cobas TaqMan assay in patients with viral loads previously undetectable by the Roche Cobas Amplicor Monitor.
Clin Infect Dis, 48 (2009), pp. 260-262
[35.]
B. Joos, M. Fischer, H. Kuster, S.K. Pillai, J.K. Wong, J. Böni, et al.
HIV rebounds from latently infected cells, rather than from continuing low-level replication.
Proc Natl Acad Sci USA, 105 (2008), pp. 16725-16730
[36.]
K. Korn, B. Weissbrich, C. Henke-Gendo, A. Heim, C.M. Jauer, N. Taylor, et al.
Singlepoint mutations causing more than 100-fold underestimation of human immunodeficiency virus type 1 (HIV-1) load with the Cobas TaqMan HIV-1 real-time PCR assay.
J Clin Microbiol, 47 (2009), pp. 1238-1240
[37.]
K.T. Troppan, E. Stelzl, D. Violan, M. Winkler, H.H. Kessler.
Evaluation of the new VERSANT HIV-1 RNA 1.0 Assay (kPCR) for quantitative detection of human immunodeficiency virus type 1 RNA.
J Clin Virol, 46 (2009), pp. 69-74
[38.]
A. Garcia-Diaz, G.S. Clewley, C.L. Booth, W. Labett, N. McAllister, A.M. Geretti.
Comparative evaluation of the performance of the Abbott real-time human immunodeficiency virus type 1 (HIV-1) assay for measurement of HIV-1 plasma viral load following automated specimen preparation.
J Clin Microbiol, 44 (2006), pp. 1788-1791
[39.]
G.L. Davis.
Monitoring of viral levels during therapy of hepatitis C.
Hepatology, 36 (2002), pp. S145-S151
[40.]
T. Berg, C. Sarrazin, E. Herrmann, H. Hinrichsen, T. Gerlach, R. Zachoval, et al.
Prediction of treatment outcome in patients with chronic hepatitis C: Significance of baseline parameters and viral dynamics during therapy.
Hepatology, 37 (2003), pp. 600-609
[41.]
M.G. Ghany, D.B. Strader, D.L. Thomas, B. Seeff.
Diagnosis, management, and treatment of hepatitis C: an update.
Hepatology, 49 (2009), pp. 1335-1374
[42.]
J.M. Pawlotsky.
Molecular diagnosis of viral hepatitis.
Gastroenterology, 122 (2002), pp. 1554-1568
[43.]
M. Schutten.
Comparison of the Abbott Realtime HIV-1 and HCV viral load assays with commercial competitor assays.
Expert Rev Mol Diagn, 8 (2008), pp. 369-377
[44.]
J. Vermehren, A. Kau, B.C. Gärtner, R. Göbel, S. Zeuzem, C. Sarrazin.
Differences between two real-time PCR-based hepatitis C virus (HCV) assays (RealTime HCV and Cobas AmpliPrep/Cobas TaqMan) and one signal amplification assay (Versant HCV RNA 3.0) for RNA detection and quantification.
J Clin Microbiol, 46 (2008), pp. 3880-3891
[45.]
N. Orta, M.R. Guna, J.C. Latorre, J.L. Pérez, C. Gimeno.
Análisis de resultados del Programa Externo de Control de Calidad SEIMC de carga viral del VIH-1 y del VHC. Año 2006.
Enferm Infecc Microbiol Clin, 25 (2007), pp. S8-S13
[46.]
N. Orta, M.R. Guna, J.C. Latorre, M. Ovies, J.L. Pérez, C. Gimeno.
Análisis de resultados del Programa Externo de Control de Calidad SEIMC de carga viral del VIH- 1 y del VHC. Año 2007.
Enferm Infecc Microbiol Clin, 26 (2008), pp. S8-S13
[47.]
Programa Externo de Control de Calidad SEIMC. Análisis del control de carga viral VHC. Año 2008. Disponible en: http://seimc.org/control/.
[48.]
A.M. Caliendo, A. Valsamakis, Y. Zhou, B. Yen-Lieberman, J. Andersen, S. Young, et al.
Multilaboratory comparison of hepatitis C virus viral load assays.
J Clin Microbiol, 44 (2006), pp. 1726-1732
[49.]
P. Colson, A. Motte, C. Tamalet.
Broad differences between the COBAS Ampliprep total nucleic acid isolation-COBAS TaqMan 48 hepatitis C virus (HCV) and COBAS HCV Monitor v2.0 assays for quantification of serum HCV RNA of non-1 genotypes.
J Clin Microbiol, 44 (2006), pp. 1602-1603
[50.]
C. Sarrazin, B.C. Gärtner, D. Sizmann, R. Babiel, U. Mihm, W.P. Hofmann, et al.
Comparison of conventional PCR with real-time PCR and branched DNA-based assays for hepatitis C virus RNA quantification and clinical significance for genotypes 1 to 5.
J Clin Microbiol, 44 (2006), pp. 729-737
[51.]
E. Tuaillon, A.M. Mondain, L. Ottomani, L. Roudière, P. Perney, M.C. Picot, et al.
Impact of hepatitis C virus (HCV) genotypes on quantification of HCV RNA in serum by COBAS AmpliPrep/COBAS TaqMan HCV test, Abbott HCV RealTime assay, and VERSANT HCV RNA Assay.
J Clin Microbiol, 45 (2007), pp. 3077-3081
[52.]
D. Sizmann, C. Boeck, J. Boelter, D. Fischer, M. Miethke, S. Nicolaus, et al.
Fully automated quantification of hepatitis C virus (HCV) RNA in human plasma and human serum by the COBAS AmpliPrep/COBAS TaqMan system.
J Clin Virol, 38 (2007), pp. 326-333
[53.]
S. Chevaliez, M. Bouvier-Alias, R. Brillet, J.M. Pawlostky.
Overestimation and underestimation of hepatitis C virus RNA levels in a widely used real-time polymerase chain reaction-based method.
Hepatology, 46 (2007), pp. 22-31
[54.]
M.T. Pyne, E.Q. Konnick, A. Phansalkar, D.R. Hillyard.
Evaluation of the Abbott investigational use only RealTime hepatitis C virus (HCV) assay and comparison to the Roche TaqMan HCV analyte-specific reagent assay.
J Clin Microbiol, 47 (2009), pp. 2872-2878
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