Detection and quantification of Epstein-Barr virus EBER1 in EBV-infected cells by fluorescent in situ hybridization and flow cytometry

Raymond P. Stowe, Michael L. Cubbage, Clarence F. Sams, Duane L. Pierson, Alan Barrett

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A rapid and highly sensitive fluorescent in situ hybridization (FISH) assay was developed to detect Epstein-Barr virus (EBV)-infected cells in peripheral blood. Multiple fluorescein-labeled antisense oligonucleotide probes were designed to hybridize to the EBER1 transcript, which is highly expressed in latently infected cells. After a rapid (30 min) hybridization, the cells were analyzed by flow cytometry. EBER1 was detected in several positive control cell lines that have variable numbers of EBV genome copies. No EBER1 was detected in two known EBV-negative cell lines. Northern blot analyses confirmed the presence and quantity of EBER1 transcripts in each cell line. This method was used to quantify the number of EBV-infected cells in peripheral blood from a patient with chronic mononucleosis. These results indicate that EBV-infected cells can be detected at the single cell level, and that this assay can be used to quantify the number of EBV-infected cells in clinical samples. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)83-91
Number of pages9
JournalJournal of Virological Methods
Volume75
Issue number1
DOIs
StatePublished - Nov 1998

Fingerprint

Fluorescence In Situ Hybridization
Human Herpesvirus 4
Flow Cytometry
Cell Line
Oligonucleotide Probes
Antisense Oligonucleotides
Epstein-Barr virus encoded RNA 1
Fluorescein
Northern Blotting
Genome

Keywords

  • Epstein-Barr virus
  • Flow cytometry
  • Fluorescent in situ hybridization
  • Herpesvirus

ASJC Scopus subject areas

  • Virology

Cite this

Detection and quantification of Epstein-Barr virus EBER1 in EBV-infected cells by fluorescent in situ hybridization and flow cytometry. / Stowe, Raymond P.; Cubbage, Michael L.; Sams, Clarence F.; Pierson, Duane L.; Barrett, Alan.

In: Journal of Virological Methods, Vol. 75, No. 1, 11.1998, p. 83-91.

Research output: Contribution to journalArticle

Stowe, Raymond P. ; Cubbage, Michael L. ; Sams, Clarence F. ; Pierson, Duane L. ; Barrett, Alan. / Detection and quantification of Epstein-Barr virus EBER1 in EBV-infected cells by fluorescent in situ hybridization and flow cytometry. In: Journal of Virological Methods. 1998 ; Vol. 75, No. 1. pp. 83-91.
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