Detection of Epstein-Barr virus by in situ hybridization. Progress toward development of a nonisotopic diagnostic test

R. Bashir, F. Hochberg, Robert H. Singer

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

This work presents some initial quantitation of an in situ hybridization method for detection of Epstein-Barr (EB) virus nucleic acids. The purpose is to develop evaluative criteria for diagnosis of viral presence in clinical tissue specimens. In this work simultaneous denaturation of probe and target DNA and an alkaline phosphatase conjugate to detect biotinated probe were used as described by Unger et al.28 For evaluation of the hybridization, a variety of cell lines, both productively and latently infected, that were hybridized in situ using nick translated 32P-labeled viral probe sequences and counted by scintillation after the method of Lawrence and Singer were used.23 Producer cells (B95-8) showed intense foci of staining in approximately 5% of cells, with most of the other cells showing varying staining intensity. Raji cells showed varying amounts of signal from cell to cell. Namalwa cells exhibited one spot in most cells that was decreased after cells were treated with Actinomycin D (dactinomycin, Merck Sharp and Dohme, West Point, PA). Signal was identified in only a third of these same cells after sectioning. EB virus-negative Ramos cells showed no signal. The nuclear punctate nature of the signal generated is diagnostic of infected cells, and may be a useful test for cultured cells or pathologic specimens.

Original languageEnglish (US)
Pages (from-to)1035-1044
Number of pages10
JournalAmerican Journal of Pathology
Volume135
Issue number6
StatePublished - 1989
Externally publishedYes

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Human Herpesvirus 4
Routine Diagnostic Tests
In Situ Hybridization
Dactinomycin
Staining and Labeling
DNA Probes
Nucleic Acids
Alkaline Phosphatase
Cultured Cells
Cell Line

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Detection of Epstein-Barr virus by in situ hybridization. Progress toward development of a nonisotopic diagnostic test. / Bashir, R.; Hochberg, F.; Singer, Robert H.

In: American Journal of Pathology, Vol. 135, No. 6, 1989, p. 1035-1044.

Research output: Contribution to journalArticle

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