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)
JournalJournal of visualized experiments : JoVE
Issue number89
DOIs
StatePublished - 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

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(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 : JoVE, No. 89, 2014.

Research output: Contribution to journalArticle

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AU - Chang, Hsim Y.

AU - Zhao, Zijin

AU - Fan, Rong

AU - Chen, Shaoxiong

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AU - Cheng, Liang

AU - Lin, Jingmei

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