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

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, December 2003, p. 5488-5491, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5488-5491.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Human Herpesvirus 8 Load in Matched Serum and Plasma Samples of Patients with AIDS-Associated Kaposi's Sarcoma

Abeltje M. Polstra,^1* Remco van den Burg,¹ Jaap Goudsmit,^2,3 and Marion Cornelissen¹

Department of Human Retrovirology University of Amsterdam,¹ Center of Poverty-Related Communicable Diseases, Amsterdam,² Crucell Holland B.V., Leiden, The Netherlands³

Received 13 May 2003/ Returned for modification 19 August 2003/ Accepted 21 September 2003

Human herpesvirus 8 (HHV-8) (or Kaposi's sarcoma [KS]-associated herpesvirus) is associated with all forms of KS. HHV-8 DNA load in peripheral blood mononuclear cells (PBMCs) of KS patients has been shown to correlate with the clinical stage of the disease. Studies have been done to assess the HHV-8 viral load in different sample types from KS patients and its clinical relevance. This paper describes the design and evaluation of a quantitative real-time (TaqMan) PCR assay for routine diagnosis of HHV-8 infection. The linear dynamic range was 5 to 5 x 10⁶ copies of HHV-8 DNA (r² > 0.99). The assay is very sensitive, specific, and easily reproducible (less than 2% variability) and can be used for different clinical samples, such as serum, plasma, and PBMCs. The question of which clinical sample, serum or plasma, is preferable for HHV8 DNA testing was addressed, using this newly developed real-time PCR assay. From 85 patients with diagnosed AIDS-KS, matched plasma and serum samples were collected. Of the 85 patients tested, 35 were positive for HHV-8 DNA in both plasma and serum (41%), 8 were positive in serum but not plasma, and 7 had detectable HHV-8 DNA only in plasma. The HHV-8 load was similar in both plasma and serum, and no significant difference was found. However, more inhibition was seen in the plasma samples with the use of a system quality control, seal herpesvirus type 1. Therefore, our results suggest that serum is the preferred material for HHV-8 load testing, since there is less possible hindrance in the amplification than with plasma.

---

* Corresponding author. Mailing address: Department of Human Retrovirology, University of Amsterdam, Amsterdam Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31-20-5667859. Fax: 31-20-5669064. E-mail: a.m.polstra{at}amc.uva.nl.

---

Journal of Clinical Microbiology, December 2003, p. 5488-5491, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5488-5491.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Sugita, S, Shimizu, N, Watanabe, K, Mizukami, M, Morio, T, Sugamoto, Y, Mochizuki, M (2008). Use of multiplex PCR and real-time PCR to detect human herpes virus genome in ocular fluids of patients with uveitis. Br J Ophthalmol 92: 928-932 [Abstract] [Full Text]  
• Gentile, G., Capobianchi, A., Volpi, A., Palu, G., Pica, F., Calistri, A., Biasolo, M. A., Martino, P. (2005). Human Herpesvirus 8 DNA in Serum During Seroconversion in Allogeneic Bone Marrow Transplant Recipients. JNCI J Natl Cancer Inst 97: 1008-1011 [Abstract] [Full Text]