Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53

Lisa M Rimsza, Elizabeth E. Vela, Yvette M. Frutiger, Lynne C. Richter, Thomas M. Grogan, William T. Bellamy

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurrence of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3′-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings.

Original languageEnglish (US)
Pages (from-to)291-296
Number of pages6
JournalMolecular Diagnosis
Volume1
Issue number4
StatePublished - 1996

Fingerprint

Cytomegalovirus
In Situ Hybridization
Immunohistochemistry
Cytomegalovirus Infections
Viral Cytopathogenic Effect
Coronary Restenosis
Nitroblue Tetrazolium
Digoxigenin
Proteins
Streptavidin
Avidin
Microwaves
Alkaline Phosphatase
Anti-Idiotypic Antibodies
Coronary Vessels
Color
Staining and Labeling
Antigens

Keywords

  • Cytomegalovirus
  • Immunohistochemistry
  • In situ hybridization
  • Oncogene
  • P53

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Rimsza, L. M., Vela, E. E., Frutiger, Y. M., Richter, L. C., Grogan, T. M., & Bellamy, W. T. (1996). Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53. Molecular Diagnosis, 1(4), 291-296.

Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53. / Rimsza, Lisa M; Vela, Elizabeth E.; Frutiger, Yvette M.; Richter, Lynne C.; Grogan, Thomas M.; Bellamy, William T.

In: Molecular Diagnosis, Vol. 1, No. 4, 1996, p. 291-296.

Research output: Contribution to journalArticle

Rimsza, LM, Vela, EE, Frutiger, YM, Richter, LC, Grogan, TM & Bellamy, WT 1996, 'Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53', Molecular Diagnosis, vol. 1, no. 4, pp. 291-296.
Rimsza, Lisa M ; Vela, Elizabeth E. ; Frutiger, Yvette M. ; Richter, Lynne C. ; Grogan, Thomas M. ; Bellamy, William T. / Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53. In: Molecular Diagnosis. 1996 ; Vol. 1, No. 4. pp. 291-296.
@article{3f07e137c23d41f98868b17d700293f7,
title = "Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53",
abstract = "Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurrence of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3′-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings.",
keywords = "Cytomegalovirus, Immunohistochemistry, In situ hybridization, Oncogene, P53",
author = "Rimsza, {Lisa M} and Vela, {Elizabeth E.} and Frutiger, {Yvette M.} and Richter, {Lynne C.} and Grogan, {Thomas M.} and Bellamy, {William T.}",
year = "1996",
language = "English (US)",
volume = "1",
pages = "291--296",
journal = "Molecular Diagnosis and Therapy",
issn = "1177-1062",
publisher = "Adis International Ltd",
number = "4",

}

TY - JOUR

T1 - Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53

AU - Rimsza, Lisa M

AU - Vela, Elizabeth E.

AU - Frutiger, Yvette M.

AU - Richter, Lynne C.

AU - Grogan, Thomas M.

AU - Bellamy, William T.

PY - 1996

Y1 - 1996

N2 - Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurrence of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3′-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings.

AB - Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurrence of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3′-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings.

KW - Cytomegalovirus

KW - Immunohistochemistry

KW - In situ hybridization

KW - Oncogene

KW - P53

UR - http://www.scopus.com/inward/record.url?scp=29344473995&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=29344473995&partnerID=8YFLogxK

M3 - Article

VL - 1

SP - 291

EP - 296

JO - Molecular Diagnosis and Therapy

JF - Molecular Diagnosis and Therapy

SN - 1177-1062

IS - 4

ER -