This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
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 Brink, A. A. T. P.
Right arrow Articles by van den Brule, A. J. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Brink, A. A. T. P.
Right arrow Articles by van den Brule, A. J. C.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 1998, p. 3164-3169, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Nucleic Acid Sequence-Based Amplification, a New Method for Analysis of Spliced and Unspliced Epstein-Barr Virus Latent Transcripts, and Its Comparison with Reverse Transcriptase PCR

Antoinette A. T. P. Brink,1 Marcel B. H. J. Vervoort,1 Jaap M. Middeldorp,1,2 Chris J. L. M. Meijer,1 and Adriaan J. C. van den Brule1,*

Section Molecular Pathology, Department of Pathology, University Hospital Vrije Universiteit, 1007 MB Amsterdam,1 and Bioresearch Unit, Organon Teknika, 5281 RM Boxtel,2 The Netherlands

Received 20 April 1998/Returned for modification 24 June 1998/Accepted 18 August 1998

Nucleic acid sequence-based amplification (NASBA) assays were developed for direct detection of Epstein-Barr virus (EBV) transcripts encoding EBV nuclear antigen 1 (EBNA1), latent membrane proteins (LMP) 1 and 2, and BamHIA rightward frame 1 (BARF1) and for the noncoding EBV early RNA 1 (EBER1). The sensitivities of all NASBAs were at least 100 copies of specific in vitro-generated RNA. Furthermore, 1 EBV-positive JY cell in a background of 50,000 EBV-negative Ramos cells (the relative sensitivity) was detected by using the EBNA1, LMP1, and LMP2 NASBA assays. The relative sensitivity of the EBER1 NASBA was 100 EBV-positive cells, which was probably related to the loss of small RNA molecules during the isolation. The BARF1 and LMP2 NASBAs were evaluated on clinical material. BARF1 expression was found in 6 of 7 nasopharyngeal carcinomas (NPC) but in 0 of 22 Hodgkin's disease (HD) cases, whereas LMP2 expression was found in 7 of 7 NPCs and in 17 of 22 HD cases. For detection of EBNA1 transcripts in HLs (n = 12) and T- and B-cell non-Hodgkin's lymphomas (n = 3 and n = 2, respectively), NASBA was compared with reverse transcriptase (RT) PCR. Two samples were positive only with NASBA, and two other samples were positive only with RT-PCR; for all other samples, the RT-PCR and NASBA results were in agreement. We conclude that NASBA is suitable for sensitive and specific detection of the above-mentioned EBV transcripts, regardless of their splicing patterns and the presence of EBV DNA. The EBNA1, LMP2, and BARF1 NASBAs developed in this study proved to be reliable assays for detection of the corresponding transcripts in EBV-positive clinical material.

* Corresponding author. Mailing address: Section Molecular Pathology, Department of Pathology, University Hospital Vrije Universiteit, P.O. Box 7057, 1007 MB Amsterdam, The Netherlands. Phone: 31-20-4440503/44017. Fax: 31-20-4442964. E-mail: vandenbrule{at}azvu.nl.

Journal of Clinical Microbiology, November 1998, p. 3164-3169, Vol. 36, No. 11
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Seto, E., Ooka, T., Middeldorp, J., Takada, K. (2008). Reconstitution of Nasopharyngeal Carcinoma-Type EBV Infection Induces Tumorigenicity. Cancer Res. 68: 1030-1036 [Abstract] [Full Text]  
  • Pioche-Durieu, C., Keryer, C., Souquere, S., Bosq, J., Faigle, W., Loew, D., Hirashima, M., Nishi, N., Middeldorp, J., Busson, P. (2005). In Nasopharyngeal Carcinoma Cells, Epstein-Barr Virus LMP1 Interacts with Galectin 9 in Membrane Raft Elements Resistant to Simvastatin. J. Virol. 79: 13326-13337 [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]  
  • zur Hausen, A., van Beek, J., Bloemena, E., ten Kate, F. J., Meijer, C. J. L. M., van den Brule, A. J. C. (2003). No role for Epstein-Barr virus in Dutch hepatocellular carcinoma: a study at the DNA, RNA and protein levels. J. Gen. Virol. 84: 1863-1869 [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»