This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Stevens, S. J. C.
Right arrow Articles by Middeldorp, J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stevens, S. J. C.
Right arrow Articles by Middeldorp, J. M.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 2002, p. 3986-3992, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.3986-3992.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Comparison of Quantitative Competitive PCR with LightCycler-Based PCR for Measuring Epstein-Barr Virus DNA Load in Clinical Specimens

Servi J. C. Stevens, Sandra A. W. M. Verkuijlen, Adriaan J. C. van den Brule, and Jaap M. Middeldorp* Received 11 February 2002/ Returned for modification 25 May 2002/ Accepted 17 August 2002

The aim of this study was to develop a LightCycler-based real-time PCR assay for monitoring the Epstein-Barr virus (EBV) DNA load in unfractionated whole blood. This assay was compared with quantitative competitive PCR (Q-PCR) for EBV. The LightCycler-based assay was highly sensitive and reproducible when quantifying plasmid DNA in either the presence or absence of healthy donor blood DNA. Amplifying plasmid DNA in DNA backgrounds from different donors slightly increased the variation of quantification, indicating that clinical specimen DNA has an influence on quantification. In most transplant recipients, a good correlation was observed between EBV DNA load dynamics determined by LightCycler and Q-PCR in follow-up samples, although the correlation between absolute values of EBV DNA loads was weak and occasional samples were false negative in the LightCycler assay. In 253 cross-sectional blood samples from patients with Burkitt's lymphoma, infectious mononucleosis, or human immunodeficiency virus infection, a weak but significant correlation between the two methods was found (r2 = 0.37, P < 0.001). Our results indicate that the clinical specimen DNA background may influence the absolute values of EBV DNA load in LightCycler analyses but that this effect is rare. LightCycler PCR is very well suited for monitoring of EBV DNA load dynamics, and its diagnostic value is comparable to that of Q-PCR. To avoid false negativity or underestimation of viral load, future internal calibration of the LightCycler is recommended. This would also enhance EBV load assay standardization and interinstitute comparisons.

* Corresponding author. Mailing address: Department of Pathology, University Hospital Vrije Universiteit, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands. Phone: 31 20 4444001. Fax: 31 20 4442964. E-mail: j.middeldorp{at}vumc.nl.

Journal of Clinical Microbiology, November 2002, p. 3986-3992, Vol. 40, No. 11
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.11.3986-3992.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gulley, M. L., Tang, W. (2008). Laboratory Assays for Epstein-Barr Virus-Related Disease. J. Mol. Diagn. 10: 279-292 [Abstract] [Full Text]  
  • Fafi-Kremer, S., Morand, P., Barranger, C., Bargues, G., Magro, S., Bes, J., Bourgeois, P., Joannes, M., Seigneurin, J.-M. (2008). Evaluation of the Epstein-Barr Virus R-Gene Quantification Kit in Whole Blood with Different Extraction Methods and PCR Platforms. J. Mol. Diagn. 10: 78-84 [Abstract] [Full Text]  
  • Hakim, H., Gibson, C., Pan, J., Srivastava, K., Gu, Z., Bankowski, M. J., Hayden, R. T. (2007). Comparison of Various Blood Compartments and Reporting Units for the Detection and Quantification of Epstein-Barr Virus in Peripheral Blood. J. Clin. Microbiol. 45: 2151-2155 [Abstract] [Full Text]  
  • Wada, K., Kubota, N., Ito, Y., Yagasaki, H., Kato, K., Yoshikawa, T., Ono, Y., Ando, H., Fujimoto, Y., Kiuchi, T., Kojima, S., Nishiyama, Y., Kimura, H. (2007). Simultaneous Quantification of Epstein-Barr Virus, Cytomegalovirus, and Human Herpesvirus 6 DNA in Samples from Transplant Recipients by Multiplex Real-Time PCR Assay. J. Clin. Microbiol. 45: 1426-1432 [Abstract] [Full Text]  
  • Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
  • Hesselink, A. T., van den Brule, A. J. C., Groothuismink, Z. M. A., Molano, M., Berkhof, J., Meijer, C. J. L. M., Snijders, P. J. F. (2005). Comparison of Three Different PCR Methods for Quantifying Human Papillomavirus Type 16 DNA in Cervical Scrape Specimens. J. Clin. Microbiol. 43: 4868-4871 [Abstract] [Full Text]  
  • Stevens, S. J. C., Verkuijlen, S. A. W. M., Hariwiyanto, B., Harijadi, , Fachiroh, J., Paramita, D. K., Tan, I. B., Haryana, S. M., Middeldorp, J. M. (2005). Diagnostic Value of Measuring Epstein-Barr Virus (EBV) DNA Load and Carcinoma-Specific Viral mRNA in Relation to Anti-EBV Immunoglobulin A (IgA) and IgG Antibody Levels in Blood of Nasopharyngeal Carcinoma Patients from Indonesia. J. Clin. Microbiol. 43: 3066-3073 [Abstract] [Full Text]  
  • Ruiz, G., Pena, P., de Ory, F., Echevarria, J. E. (2005). Comparison of Commercial Real-Time PCR Assays for Quantification of Epstein-Barr Virus DNA. J. Clin. Microbiol. 43: 2053-2057 [Abstract] [Full Text]  
  • Simon, A., Labalette, P., Ordinaire, I., Frealle, E., Dei-Cas, E., Camus, D., Delhaes, L. (2004). Use of Fluorescence Resonance Energy Transfer Hybridization Probes To Evaluate Quantitative Real-Time PCR for Diagnosis of Ocular Toxoplasmosis. J. Clin. Microbiol. 42: 3681-3685 [Abstract] [Full Text]  
  • Wadowsky, R. M., Laus, S., Green, M., Webber, S. A., Rowe, D. (2003). Measurement of Epstein-Barr Virus DNA Loads in Whole Blood and Plasma by TaqMan PCR and in Peripheral Blood Lymphocytes by Competitive PCR. J. Clin. Microbiol. 41: 5245-5249 [Abstract] [Full Text]  
  • Shoji, N., Ohyashiki, J. H., Suzuki, A., Kubota, N., Kimura, Y., Matsubayashi, J., Mukai, K., Ohyashiki, K. (2003). Multiple Pulmonary Nodules Caused by B-Cell Post-transplant Lymphoproliferative Disorder after Bone Marrow Transplantation: Monitoring Epstein-Barr Virus Viral Load. Jpn J Clin Oncol 33: 408-412 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»