Skip to main content
SpringerLink
Log in

Quantitation of Viral Load

  • Chapter
Clinical and Forensic Applications of Capillary Electrophoresis

Part of the book series: Pathology and Laboratory Medicine ((PLM))

  • 647 Accesses

Abstract

Viruses, the world’s smallest organisms, are a large group of single cell organisms comprised of nucleic acid surrounded by a protein coat. Pathogenic viruses play an important role in many human diseases, including hepatitis, cancer, and AIDS, as well as the common cold. These viruses are of particular concern for patients who are immune compromised, those undergoing transplant procedures with donor organs, and/or those receiving blood from human sources.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Similar content being viewed by others

References

  1. Zaia, J. A., Gallez-Hawkins, G. M., Tegtmeier, B. R., ter Veer, A., Li, X., Niland, J. C., and Forman, S. J. (1997) Late cytomegalovirus disease in marrow transplantation is predicted by virus load in plasma. J. Infect. Dis. 176, 72–75.

    Google Scholar 

  2. Boivin, G., Handfield, J., Toma, E., Murray, G. Lalonde, R., and Bergeron, M. G. (1998) Comparative evaluation of the cytomegalovirus DNA load in polymorphonuclear leukocytes and plasma of human immunodeficiency virus-infected subjects. J. Infect. Dis. 177, 355–360.

    Google Scholar 

  3. Cope, A. V., Sabin, C., Burroughs, A., Rolles, K., Griffiths, P. D., and Emery, V. C. (1997) Interrelationships among quantity of human cytomegalovirus (HCMV) DNA in blood, donor-recipient serostatus, and administration of methylprednisolone as risk factors for HCMV disease following liver transplantation. J. Infect. Dis. 176, 1484–1490.

    Google Scholar 

  4. Boeckh, M. and Boivin, G. (1998) Quantitation of cytomegalovirus: methodologic aspects and clinical applications. Clin. Microbio. Rev. 11, 533–554.

    Google Scholar 

  5. Chatellard, P., Sahli, R., Iten, A., von Overbeck, J., and Meylan, P. R. (1998) Single tube competitive PCR for quantitation of CMV DNA in the blood of HIV+ and solid organ transplant patients. J. Virol. Meth. 71, 137–146.

    Google Scholar 

  6. Gerna, G., Sarasini, A., Baldanti, F., Percivalle, E., Zella, D., and Revello, M. G. (1997) Quantitative systemic and local evaluation of the antiviral effect of ganciclovir and foscarnet induction treatment on human cytomegalovirus gastrointestinal disease of patients with AIDS. Italian Foscarnet GID Study Group. Antiviral Res. 34, 39–50.

    Google Scholar 

  7. Lazzarotto, T., Campisi, T., Dal Monte, P., Galli, S., Spezzacatena, P., Guglielmi, P., and Landini, M. P. (1996) A quantitative test ( HCMV-hybridcapture(TM)) to detect human cytomegalovirus DNA in the blood of immunocompromised patients compared with antigenemia and polymerase chain reaction. New Microbiol. 19, 193–201.

    Google Scholar 

  8. Poirier-Toulemonde, A. S., Imbert-Marcille, B. M., Ferre-Aubineau, V., Besse,B., Le Roux, M. G., Cantarovich, D., and Billaudel, S. (1997) Successful quantification of cytomegalovirus DNA by competitive PCR and detection with capillary electrophoresis. Mol. Cell. Probes. 1 l, 11–23.

    Google Scholar 

  9. Boivin, G., Gilbert, C., Morissette, M., Handfield, J., Goyette, N., and Bergeron, M. G. (1997) A case of ganciclovir-resistant cytomegalovirus (CMV) retinitis in a patient with AIDS: longitudinal molecular analysis of the CMV viral load and viral mutations in blood compartments. AIDS 11, 867–873.

    Article  PubMed  CAS  Google Scholar 

  10. Gerna, G., Percivalle, E., Baldanti, F., Sarasini, A., Zavattoni, M., Furione, M., et al. (1998) Diagnostic significance and clinical impact of quantitative assays for diagnosis of human cytomegalovirus infection/disease in immunocompromised patients. New Microbiol. 21, 293–308.

    PubMed  CAS  Google Scholar 

  11. Hebart, H., Schroder, A., Loffler, J., Klingebiel, T., Martin, H., Wassmann, B., et al. (1997) Cytomegalovirus monitoring by polymerase chain reaction of whole blood samples from patients undergoing autologous bone marrow or peripheral blood progenitor cell transplantation. J. Infect. Dis. 175, 1490–1493.

    Google Scholar 

  12. de Jong, M. D., Boucher, C. A., Danner, S. A., Gazzard, B., Griffiths, P. D., Katlama,C., et al. (1998) Summary of the international consensus symposium on management of HIV, CMV and hepatitis virus infections. Antiviral Res. 37, 1–16.

    Google Scholar 

  13. Evans, A. A., O’Connell, A. P., Pugh, J. C., Mason, W. S., Shen, F. M., Chen, G. C., et al. (1998) Geographic variation in viral load among hepatitis B carriers with differing risks of hepatocellular carcinoma. Cancer Epidemiol. Biomarkers Prevention 7, 559–565.

    Google Scholar 

  14. Huang, X., , M. J., and Zare, R. N. (1988) Bias in quantitative capillary zone electrophoresis caused by electrokinetic sample injection. Anal. Chem. 60, 375–377.

    Google Scholar 

  15. Simpson, P. R., Yu, X. H., Redza, Z. M., Anson, J. G., Chan, S. H., and Lin, Y. (1997) Quantification of hepatitis B virus DNA using competitive PCR and a scintillation proximity assay. J. Virol. Methods 69, 197–208.

    Google Scholar 

  16. Dienstag, J. L., Perrillo, R. P., Schiff, E. R., Bartholomew, M., Vicary, C., and Rubin, M. (1995) A preliminary trial of lamivudine for chronic hepatitis B. New Engl. J. Med. 333, 1657–1661.

    Google Scholar 

  17. Perelson, A. S., Neumann, A. U., Markowitz, M., Leonard, J. M., and Ho, D. D. (1996) HIV-1 dynamics in vivo: virion clearance rate, infected cell life-span, and viral generation time. Science 271, 1582–1586.

    Article  PubMed  CAS  Google Scholar 

  18. Piatak, M., Saag, M. S., and Yang, L. C. (1993) High levels of HIV in plasma during all stages of infection determined by competitive PCR. Science 259, 1749–1754.

    Article  PubMed  CAS  Google Scholar 

  19. Ho, D. D., Neumann, A. U., Perelson, A. S., Chen, W., Leonard, J. M., and Markowitz, M. (1995) Rapid turnover of plasma virions and CD4 sup + lymphocytes in HIV-1 infection. Nature 373, 123–126.

    Article  PubMed  CAS  Google Scholar 

  20. Marschner, I. C., Collier, A. C., and Coombs, R. W. (1998) Use of changes in plasma levels of human immunodeficiency virus type 1 RNA to assess clinical benefit of antiretroviral therapy. J. Infect. Dis. 177, 40–47.

    Google Scholar 

  21. Mellors, J. W., Munoz, A. M., and Giorgi, J. V. (1997) Plasma viral load and CD4 sup + lymphocytes as prognostic markers of HIV-1 infection. Ann. Intern. Med. 126, 946–954.

    Google Scholar 

  22. Montaner, J., DeMasi, R., and Hill, A. (1997) Validation of HIV-1 RNA and CD4 count as surrogate markers in the CAESAR trial, in: Program and abstracts of the 6th European Conference on Clinical Aspects and Treatment of HIV Infection; October 11–15, 1997. Hamburg, Germany, Abstract 207.

    Google Scholar 

  23. Wei, X., Ghosh, S. K., and Taylor, M. E. (1995) Viral dynamics in HIV-1 infection. Nature 373, 117–122.

    Article  PubMed  CAS  Google Scholar 

  24. Carpenter, C. C. J., Fischl, M. A., and Hammer, S. M. (1997) Antiretroviral therapy for HIV infection in 1997: updated recommendations of the International AIDS Society-USA panel. JAMA 277, 1962–1969.

    Article  PubMed  CAS  Google Scholar 

  25. US Department of Health and Human Services and the Henry J. Kaiser Family Foundation. (1998) Guidelines for the use of antiretroviral agents in HIV-1 infected adults and adolescents. MMWR Morb. Mortal. Wkly. Rep. 47 (RR-05), 43–82.

    Google Scholar 

  26. Brandsma, J. and Miller, G. (1980) Nucleic acid spot hybridization: rapid quantitative screening of lymphoid cell lines for Epstein-Barr virus. Proc. Nati. Acad. Sci. USA 77, 6851–6855.

    Google Scholar 

  27. Ferre, F., Marchese, A., and Pezzoli, P. (1994) Quantitative PCR: an overview, in PCR. The Polymerase Chain Reaction ( Mullis, K. B., Ferre, F., and Gibbs, R. A., eds.), Birkhauser, Boston, MA.

    Google Scholar 

  28. Kolesar, J. M., Allen, P. A., and Doren, C. M. (1997) Direct quantification of HIV-1 RNA by CE-LIF. J. Chrom. B 697, 189–194.

    Article  CAS  Google Scholar 

  29. Dieguez-Lucena, J. L., Ruiz-Galdon, M., Morell-Ocana, M., Garcia-Villanova, J., Flores-Polanco, F. J., and Reyes-Engel, A. (1994) Capillary electrophoresis compared with silver staining of polyacrylamide gels for quantification of per products. Clin. Chem. 40, 493, 494.

    Google Scholar 

  30. Haupt, W. and Pingoud, A. (1983) Comparison of several high performance liquid chromatography techniques for the separation of oligodeoxynucleotides according to their chain lengths. J. Chromatogr. 260, 419–423.

    Article  CAS  Google Scholar 

  31. Oefner, P. J., Bonn, G. K., Huber, C. G., and Nathakarnkitkoll, S. (1992) Comparative study of capillary zone electrophoresis and high performance liquid chromatography in the analysis of oligonucleotides and DNA. J. Chromatogr. 625, 331–340.

    Article  PubMed  CAS  Google Scholar 

  32. Karger, B. L., Chu, Y. H., and Foret, F. (1995) Capillary electrophoresis of proteins and nucleic acids. Annu. Rev. Biophys. Biomol. Struct. 24, 579–610.

    Google Scholar 

  33. Kuhr, W. G. and Monnig, C. A. (1992) Fundamental reviews: capillary electrophoresis. Anal. Chem. 62, 403R - 414R.

    Article  Google Scholar 

  34. Landers, J. P. (1995) Clinical capillary electrophoresis. Clin. Chem. 41, 495–509.

    Google Scholar 

  35. Gelfi, C., Leoncini, F., Righetti, P. G., Cremonsei, L., di Blasio, A. M., Carniti, C., and Vignali, M. (1995) Separation and quantitation of reverse transcriptase polymerase chain reaction fragments of basic fibroblast growth factor by capillary electrophoresis in polymer networks. Electrophoresis 16, 780–783.

    Article  PubMed  CAS  Google Scholar 

  36. Lu, W., Han, D. S., Yuan, J., and Andrieu, J. M. (1994) Multi-target PCR analysis by capillary electrophoresis and laser induced fluorescence. Nature 368, 269–271.

    Article  PubMed  CAS  Google Scholar 

  37. Rossomando, E. F., White, L., and Ulfelder, K. J. (1994) Capillary electrophoresis: separation and quantitation of reverse transcriptase polymerase chain reaction products from polio virus. J. Chromatogr. B 656, 159–168.

    Article  CAS  Google Scholar 

  38. Schwartz, H. E. and Ulfelder, K. J. (1991) Analysis of DNA restriction fragments and polymerase chain reaction products towards detection of the AIDS (HIV-1) virus in blood. J. Chromatogr. 559, 267–283.

    Article  PubMed  CAS  Google Scholar 

  39. Stalbom, B. M., Torven, A., and Lundberg, L. G. (1994) Application of capillary electrophoresis to the post polymerase chain reaction analysis of rat mRNA for gastric H+, K+-ATPase. Ann. Biochem. 217, 91–97.

    Google Scholar 

  40. Schwartz, H. E. and Ulfelder, K. J. (1992) Capillary electrophoresis with laser induced fluorescence detection of PCR fragments using thiazole orange. Ann. Chem. 64, 1737–1740

    Google Scholar 

  41. Schwarz, H. E., Ulfelder, K. J., Chen, F. T. A., and Pentoney, S. L. (1994) The utility of laser induced fluorescence detection in applications of capillary electrophoresis. J. Cap. Electrophor. 1, 36–54.

    Google Scholar 

  42. Srinivasan, K., Girard, J. E., and Williams, P. (1993) Electrophoretic separations of polymerase chain reaction-amplified DNA fragments in DNA typing using a capillary electrophoresis-laser induced fluorescence system. J. Chromatogr. 652, 83–91.

    Article  CAS  Google Scholar 

  43. Srinivasan, K., Morris, S. C., and Girard, J. E. (1993) Enhanced detection of PCR products through use of TOTO and YOYO intercalating dyes with laser induced fluorescence-capillary electrophoresis. Appi. Theoret. Electrophor. 3, 235–239.

    Google Scholar 

  44. Hjerten, S. (1985) High performance electrophoresis: elimination of electroendosmosis and solute absorption. J. Chromatogr. 347: 191–198.

    Article  CAS  Google Scholar 

  45. Schmalzing, D., Piggee, C. A., Foret, F., Carrilho, E., and Karger, B. L. (1993) Characterization and performance of a neutral hydrophilic coating for the capillary electrophoretic separation of biopolymers. J. Chromatogr. A 652, 149–159.

    Article  PubMed  CAS  Google Scholar 

  46. Tsuda, T. (1987) Modification of electroosmotic flow with cetyltrimethylammonium bromide in capillary electrophoresis. J. High Resolut. Chromatogr. 10, 622–624.

    Google Scholar 

  47. Hwang, S. J., Leem S. D., Lu, R. H., Chan, C. Y., Lai, L., Co, R. L., and Tong, M. J. (1996) Comparison of three different hybridization assays in the quantitative measurement of serum hepatitis B virus DNA. J. Virol. Methods 62, 123–129.

    Google Scholar 

  48. Kleparnik, K., Garner, M., and Bocek, P. (1995) Injection bias of DNA fragments in capillary electrophoresis with seiving. J. Chromatogr. A 698, 375–383.

    Article  PubMed  CAS  Google Scholar 

  49. Heller, C. (1995) Capillary electrophoresis of proteins and nucleic acids in gels and entangled polymer solutions. J. Chromatogr. A 698, 19–31.

    Article  CAS  Google Scholar 

  50. Pariat, Y. F., Berka, J., Heiger, D. N., Schmitt, T., and Vilenchik, M. (1993) Separation of DNA fragments by capillary electrophoresis using replaceable linear polyacrylamide matrices. J. Chromatogr. A 652, 57–66.

    Article  PubMed  CAS  Google Scholar 

  51. Kolesar, J. M., Rizzo, J. D., Kuhn, J. G. (1995) Quantitative analysis of NQO1 gene expression by RT-PCR and CE-LIF. J. Cap. Electrophor. 2, 287–290.

    Google Scholar 

  52. Kunkel, A., Degenhardt, M., and Watzig, H. (1996) Precise quantitative results by capillary electrophoresis (CE): instrumental aspects (an update) Proceedings from the 8th International Symposium on High Performance Capillary Electrophoresis 17, 140 (Abstract).

    Google Scholar 

  53. Butler, J. M., McCord, B. R., and Jung, J. M. (1994) Quantitation of polymerase chain reaction products by capillary electrophoresis using laser fluorescence. J. Chromatogr. B 658, 271–280.

    Article  CAS  Google Scholar 

  54. Fasco, M. J., Treanor, C. P., Spivack, S., Figge, H. L., and Kaminsky, L. S. (1995) Quantitative RNA-polymerase chain reaction-DNA analysis by capillary electrophoresis with laser induced fluorescence. Anal. Biochem. 224, 140–147.

    Google Scholar 

  55. Glasel, J. (1995) Validity of nucleic acid purities monitored by 260nm/280nm absorbance ratios. Biotechniques 18, 62, 63.

    Google Scholar 

  56. Kuypers, A., Meijerink, J. P., Smetsers, T., Linssen, P., and Mensink, E. (1994) Quantitative analysis of DNA aberrations amplified by competitive polymerase chain reaction using capillary electrophoresis. J. Chromatogr. B 660, 271–277.

    Article  CAS  Google Scholar 

  57. Kolesar, J. M., Burris, H., and Kuhn, J. G. (1995) Detection of a point mutation in NQO1 (DT-diaphorase) in a patient with colon cancer. J. Natl. Cancer Inst. 87, 1022–1024.

    Google Scholar 

  58. Khrapko, K., Hanekamp, J. S., Thilly, W. G., Belenkii, A., Foret, F., and Karger, B. L. (1994) Constant denaturant capillary electrophoresis (CDCE): a high resolution approach to mutational analysis. Nucleic Acids Res. 22, 364–369.

    Article  PubMed  CAS  Google Scholar 

  59. Kumar, R., Hanekamp, J. S., Louhelainen, K., Burvall, K., Onfelt, A., Hemminki, K., and Thilly, W. G. (1995) Separation of transforming amino acid-substituting mutations in codons 12, 13, and 61 of the n-ras gene by constant denaturant capillary electrophoresis ( CDCE ). Carcinogenesis 16, 2667–2673.

    Google Scholar 

  60. Mitchell, C. E., Belinsky, S. A., and Lechner, J. F. (1995) Detection and quantitation of mutant K-ras codon 12 restriction fragments by capillary electrophoresis. Ann. Biochem. 224, 148–153.

    Google Scholar 

Download references

Authors
  1. Jill M. Kolesar
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Texas Medical Branch, Galveston, TX, USA

    John R. Petersen  & Amin A. Mohammad  & 

Rights and permissions

Reprints and permissions

Copyright information

© 2001 Springer Science+Business Media New York

About this chapter

Cite this chapter

Kolesar, J.M. (2001). Quantitation of Viral Load. In: Petersen, J.R., Mohammad, A.A. (eds) Clinical and Forensic Applications of Capillary Electrophoresis. Pathology and Laboratory Medicine. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59259-120-6_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-59259-120-6_13

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61737-113-4

  • Online ISBN: 978-1-59259-120-6

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation