Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies

Lisa M Rimsza, Elizabeth E. Vela, Yvette M. Frutiger, Catherine S. Rangel, Max Solano, Lynne C. Richter, Thomas M. Grogan, William T. Bellamy

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The authors questioned whether an automated kinetic mode assay of combined cytomegalovirus (CMV) late viral message and immediate and early antigens might result in a more sensitive and timely CMV diagnosis relevant to speedy treatment in the transplant setting. Toward this end, two cohorts were studied using automated in situ hybridization (ISH) for CMV as well as immunohistochemistry (IHC). The first cohort of patients consisted of 19 cases that were histologically positive (CMV-associated cytopathic change). A second cohort consisted of 10 cases that were histologically negative, yet culture positive. From the first cohort of histologically positive cases, 100% were positive by both ISH and IHC run on separate slides. In the second cohort, CMV was detected overall in 70% of cases (50% by ISH alone and 30% by IHC alone). These results indicate that a combined assay of ISH and IHC can detect more cases than routine hematoxylin and eosin staining or either assay alone. In two illustrative cases, used to demonstrate the feasibility of combining ISH and IHC, the authors used a combined two-color assay (ISH and IHC) performed sequentially on the same slide. The combined assays resulted in colocalized single cell message and protein in some cells and demonstrated more positive cells overall (some positive by IHC alone, some by ISH alone, and some by both) than either assay alone. The combined dual color assay can be completed within 4 to 5 hours giving the prospect of a same day result, which is faster than shell vial technique with immunofluorescence (24 to 48 hours) or culture (7 to 14 days). This study demonstrates that combining CMV message and protein assays results in a more sensitive assay and, when carried out in the kinetic mode, allows a speedy result relevant to early anti-CMV therapy.

Original languageEnglish (US)
Pages (from-to)544-548
Number of pages5
JournalAmerican Journal of Clinical Pathology
Volume106
Issue number4
StatePublished - Oct 1996

Fingerprint

Cytomegalovirus
Paraffin
In Situ Hybridization
Immunohistochemistry
Biopsy
Color
Hematoxylin
Eosine Yellowish-(YS)
Fluorescent Antibody Technique
Staining and Labeling
Transplants
Antigens
Therapeutics
Proteins

Keywords

  • CMV
  • Immunohistochemistry
  • In Situ Hybridization

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies. / Rimsza, Lisa M; Vela, Elizabeth E.; Frutiger, Yvette M.; Rangel, Catherine S.; Solano, Max; Richter, Lynne C.; Grogan, Thomas M.; Bellamy, William T.

In: American Journal of Clinical Pathology, Vol. 106, No. 4, 10.1996, p. 544-548.

Research output: Contribution to journalArticle

Rimsza, Lisa M ; Vela, Elizabeth E. ; Frutiger, Yvette M. ; Rangel, Catherine S. ; Solano, Max ; Richter, Lynne C. ; Grogan, Thomas M. ; Bellamy, William T. / Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies. In: American Journal of Clinical Pathology. 1996 ; Vol. 106, No. 4. pp. 544-548.
@article{5decdb16b12c4c25901d3afc45df8c10,
title = "Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies",
abstract = "The authors questioned whether an automated kinetic mode assay of combined cytomegalovirus (CMV) late viral message and immediate and early antigens might result in a more sensitive and timely CMV diagnosis relevant to speedy treatment in the transplant setting. Toward this end, two cohorts were studied using automated in situ hybridization (ISH) for CMV as well as immunohistochemistry (IHC). The first cohort of patients consisted of 19 cases that were histologically positive (CMV-associated cytopathic change). A second cohort consisted of 10 cases that were histologically negative, yet culture positive. From the first cohort of histologically positive cases, 100{\%} were positive by both ISH and IHC run on separate slides. In the second cohort, CMV was detected overall in 70{\%} of cases (50{\%} by ISH alone and 30{\%} by IHC alone). These results indicate that a combined assay of ISH and IHC can detect more cases than routine hematoxylin and eosin staining or either assay alone. In two illustrative cases, used to demonstrate the feasibility of combining ISH and IHC, the authors used a combined two-color assay (ISH and IHC) performed sequentially on the same slide. The combined assays resulted in colocalized single cell message and protein in some cells and demonstrated more positive cells overall (some positive by IHC alone, some by ISH alone, and some by both) than either assay alone. The combined dual color assay can be completed within 4 to 5 hours giving the prospect of a same day result, which is faster than shell vial technique with immunofluorescence (24 to 48 hours) or culture (7 to 14 days). This study demonstrates that combining CMV message and protein assays results in a more sensitive assay and, when carried out in the kinetic mode, allows a speedy result relevant to early anti-CMV therapy.",
keywords = "CMV, Immunohistochemistry, In Situ Hybridization",
author = "Rimsza, {Lisa M} and Vela, {Elizabeth E.} and Frutiger, {Yvette M.} and Rangel, {Catherine S.} and Max Solano and Richter, {Lynne C.} and Grogan, {Thomas M.} and Bellamy, {William T.}",
year = "1996",
month = "10",
language = "English (US)",
volume = "106",
pages = "544--548",
journal = "American Journal of Clinical Pathology",
issn = "0002-9173",
publisher = "American Society of Clinical Pathologists",
number = "4",

}

TY - JOUR

T1 - Rapid automated combined in situ hybridization and immunohistochemistry for sensitive detection of cytomegalovirus in paraffin-embedded tissue biopsies

AU - Rimsza, Lisa M

AU - Vela, Elizabeth E.

AU - Frutiger, Yvette M.

AU - Rangel, Catherine S.

AU - Solano, Max

AU - Richter, Lynne C.

AU - Grogan, Thomas M.

AU - Bellamy, William T.

PY - 1996/10

Y1 - 1996/10

N2 - The authors questioned whether an automated kinetic mode assay of combined cytomegalovirus (CMV) late viral message and immediate and early antigens might result in a more sensitive and timely CMV diagnosis relevant to speedy treatment in the transplant setting. Toward this end, two cohorts were studied using automated in situ hybridization (ISH) for CMV as well as immunohistochemistry (IHC). The first cohort of patients consisted of 19 cases that were histologically positive (CMV-associated cytopathic change). A second cohort consisted of 10 cases that were histologically negative, yet culture positive. From the first cohort of histologically positive cases, 100% were positive by both ISH and IHC run on separate slides. In the second cohort, CMV was detected overall in 70% of cases (50% by ISH alone and 30% by IHC alone). These results indicate that a combined assay of ISH and IHC can detect more cases than routine hematoxylin and eosin staining or either assay alone. In two illustrative cases, used to demonstrate the feasibility of combining ISH and IHC, the authors used a combined two-color assay (ISH and IHC) performed sequentially on the same slide. The combined assays resulted in colocalized single cell message and protein in some cells and demonstrated more positive cells overall (some positive by IHC alone, some by ISH alone, and some by both) than either assay alone. The combined dual color assay can be completed within 4 to 5 hours giving the prospect of a same day result, which is faster than shell vial technique with immunofluorescence (24 to 48 hours) or culture (7 to 14 days). This study demonstrates that combining CMV message and protein assays results in a more sensitive assay and, when carried out in the kinetic mode, allows a speedy result relevant to early anti-CMV therapy.

AB - The authors questioned whether an automated kinetic mode assay of combined cytomegalovirus (CMV) late viral message and immediate and early antigens might result in a more sensitive and timely CMV diagnosis relevant to speedy treatment in the transplant setting. Toward this end, two cohorts were studied using automated in situ hybridization (ISH) for CMV as well as immunohistochemistry (IHC). The first cohort of patients consisted of 19 cases that were histologically positive (CMV-associated cytopathic change). A second cohort consisted of 10 cases that were histologically negative, yet culture positive. From the first cohort of histologically positive cases, 100% were positive by both ISH and IHC run on separate slides. In the second cohort, CMV was detected overall in 70% of cases (50% by ISH alone and 30% by IHC alone). These results indicate that a combined assay of ISH and IHC can detect more cases than routine hematoxylin and eosin staining or either assay alone. In two illustrative cases, used to demonstrate the feasibility of combining ISH and IHC, the authors used a combined two-color assay (ISH and IHC) performed sequentially on the same slide. The combined assays resulted in colocalized single cell message and protein in some cells and demonstrated more positive cells overall (some positive by IHC alone, some by ISH alone, and some by both) than either assay alone. The combined dual color assay can be completed within 4 to 5 hours giving the prospect of a same day result, which is faster than shell vial technique with immunofluorescence (24 to 48 hours) or culture (7 to 14 days). This study demonstrates that combining CMV message and protein assays results in a more sensitive assay and, when carried out in the kinetic mode, allows a speedy result relevant to early anti-CMV therapy.

KW - CMV

KW - Immunohistochemistry

KW - In Situ Hybridization

UR - http://www.scopus.com/inward/record.url?scp=0029748323&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029748323&partnerID=8YFLogxK

M3 - Article

C2 - 8853045

AN - SCOPUS:0029748323

VL - 106

SP - 544

EP - 548

JO - American Journal of Clinical Pathology

JF - American Journal of Clinical Pathology

SN - 0002-9173

IS - 4

ER -