qPCR is a sensitive and rapid method for detection of cytomegaloviral DNA in formalin-fixed, paraffin-embedded biopsy tissue

Morgan H. McCoy, Kristin Post, Joyashree D. Sen, Hsim Y. Chang, Zijin Zhao, Rong Fan, Shaoxiong Chen, Diane Leland, Liang Cheng, Jingmei Lin

Research output: Contribution to journalArticle

Abstract

It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for developing severe infection. Many laboratory methods for the detection of CMV infection have been developed, including serology, viral culture, and molecular methods. Often, these methods reflect systemic involvement with CMV and do not specifically identify local tissue involvement. Therefore, detection of CMV infection in the GI tract is frequently done by traditional histology of biopsy tissue. Hematoxylin and eosin (H&E) staining in conjunction with immunohistochemistry (IHC) have remained the mainstays of examining these biopsies. H&E and IHC sometimes result in atypical (equivocal) staining patterns, making interpretation difficult. It was shown that quantitative polymerase chain reaction (qPCR) for CMV can successfully be performed on formalin-fixed, paraffin-embedded (FFPE) biopsy tissue for very high sensitivity and specificity. The goal of this protocol is to demonstrate how to perform qPCR testing for the detection of CMV in FFPE biopsy tissue in a clinical laboratory setting. This method is likely to be of great benefit for patients in cases of equivocal staining for CMV in GI biopsies.

Original languageEnglish (US)
Article numbere51570
JournalJournal of Visualized Experiments
Issue number89
DOIs
StatePublished - Sep 7 2014

Fingerprint

Biopsy
Polymerase chain reaction
Formaldehyde
Paraffin
Paraffins
DNA
Cytomegalovirus
Tissue
Cytomegalovirus Infections
Polymerase Chain Reaction
Staining and Labeling
Gastrointestinal Tract
Immunohistochemistry
Clinical laboratories
Histology
Serology
Hematoxylin
Eosine Yellowish-(YS)
Sensitivity and Specificity
Testing

Keywords

  • Biopsy
  • CMV
  • Cytomegalovirus
  • Formalin-fixed
  • Gastrointestinal
  • Genetics
  • Issue 89
  • Paraffin-embedded tissue
  • Real-time PCR
  • qPCR

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

qPCR is a sensitive and rapid method for detection of cytomegaloviral DNA in formalin-fixed, paraffin-embedded biopsy tissue. / McCoy, Morgan H.; Post, Kristin; Sen, Joyashree D.; Chang, Hsim Y.; Zhao, Zijin; Fan, Rong; Chen, Shaoxiong; Leland, Diane; Cheng, Liang; Lin, Jingmei.

In: Journal of Visualized Experiments, No. 89, e51570, 07.09.2014.

Research output: Contribution to journalArticle

@article{3c066fbd6e9b4c11b0b63e7e4526fa19,
title = "qPCR is a sensitive and rapid method for detection of cytomegaloviral DNA in formalin-fixed, paraffin-embedded biopsy tissue",
abstract = "It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for developing severe infection. Many laboratory methods for the detection of CMV infection have been developed, including serology, viral culture, and molecular methods. Often, these methods reflect systemic involvement with CMV and do not specifically identify local tissue involvement. Therefore, detection of CMV infection in the GI tract is frequently done by traditional histology of biopsy tissue. Hematoxylin and eosin (H&E) staining in conjunction with immunohistochemistry (IHC) have remained the mainstays of examining these biopsies. H&E and IHC sometimes result in atypical (equivocal) staining patterns, making interpretation difficult. It was shown that quantitative polymerase chain reaction (qPCR) for CMV can successfully be performed on formalin-fixed, paraffin-embedded (FFPE) biopsy tissue for very high sensitivity and specificity. The goal of this protocol is to demonstrate how to perform qPCR testing for the detection of CMV in FFPE biopsy tissue in a clinical laboratory setting. This method is likely to be of great benefit for patients in cases of equivocal staining for CMV in GI biopsies.",
keywords = "Biopsy, CMV, Cytomegalovirus, Formalin-fixed, Gastrointestinal, Genetics, Issue 89, Paraffin-embedded tissue, Real-time PCR, qPCR",
author = "McCoy, {Morgan H.} and Kristin Post and Sen, {Joyashree D.} and Chang, {Hsim Y.} and Zijin Zhao and Rong Fan and Shaoxiong Chen and Diane Leland and Liang Cheng and Jingmei Lin",
year = "2014",
month = "9",
day = "7",
doi = "10.3791/51570",
language = "English (US)",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "89",

}

TY - JOUR

T1 - qPCR is a sensitive and rapid method for detection of cytomegaloviral DNA in formalin-fixed, paraffin-embedded biopsy tissue

AU - McCoy, Morgan H.

AU - Post, Kristin

AU - Sen, Joyashree D.

AU - Chang, Hsim Y.

AU - Zhao, Zijin

AU - Fan, Rong

AU - Chen, Shaoxiong

AU - Leland, Diane

AU - Cheng, Liang

AU - Lin, Jingmei

PY - 2014/9/7

Y1 - 2014/9/7

N2 - It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for developing severe infection. Many laboratory methods for the detection of CMV infection have been developed, including serology, viral culture, and molecular methods. Often, these methods reflect systemic involvement with CMV and do not specifically identify local tissue involvement. Therefore, detection of CMV infection in the GI tract is frequently done by traditional histology of biopsy tissue. Hematoxylin and eosin (H&E) staining in conjunction with immunohistochemistry (IHC) have remained the mainstays of examining these biopsies. H&E and IHC sometimes result in atypical (equivocal) staining patterns, making interpretation difficult. It was shown that quantitative polymerase chain reaction (qPCR) for CMV can successfully be performed on formalin-fixed, paraffin-embedded (FFPE) biopsy tissue for very high sensitivity and specificity. The goal of this protocol is to demonstrate how to perform qPCR testing for the detection of CMV in FFPE biopsy tissue in a clinical laboratory setting. This method is likely to be of great benefit for patients in cases of equivocal staining for CMV in GI biopsies.

AB - It is crucial to identify cytomegalovirus (CMV) infection in the gastrointestinal (GI) tract of immunosuppressed patients, given their greater risk for developing severe infection. Many laboratory methods for the detection of CMV infection have been developed, including serology, viral culture, and molecular methods. Often, these methods reflect systemic involvement with CMV and do not specifically identify local tissue involvement. Therefore, detection of CMV infection in the GI tract is frequently done by traditional histology of biopsy tissue. Hematoxylin and eosin (H&E) staining in conjunction with immunohistochemistry (IHC) have remained the mainstays of examining these biopsies. H&E and IHC sometimes result in atypical (equivocal) staining patterns, making interpretation difficult. It was shown that quantitative polymerase chain reaction (qPCR) for CMV can successfully be performed on formalin-fixed, paraffin-embedded (FFPE) biopsy tissue for very high sensitivity and specificity. The goal of this protocol is to demonstrate how to perform qPCR testing for the detection of CMV in FFPE biopsy tissue in a clinical laboratory setting. This method is likely to be of great benefit for patients in cases of equivocal staining for CMV in GI biopsies.

KW - Biopsy

KW - CMV

KW - Cytomegalovirus

KW - Formalin-fixed

KW - Gastrointestinal

KW - Genetics

KW - Issue 89

KW - Paraffin-embedded tissue

KW - Real-time PCR

KW - qPCR

UR - http://www.scopus.com/inward/record.url?scp=84904204781&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84904204781&partnerID=8YFLogxK

U2 - 10.3791/51570

DO - 10.3791/51570

M3 - Article

C2 - 25046572

AN - SCOPUS:84904204781

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 89

M1 - e51570

ER -