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.
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine; momccoy@iupui.edu.
authors.forename: Morgan H
authors.initials: MH
authors.lastname: McCoy
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University Health.
authors.forename: Kristin
authors.initials: K
authors.lastname: Post
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University Health.
authors.forename: Joyashree D
authors.initials: JD
authors.lastname: Sen
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University Health.
authors.forename: Hsim Y
authors.initials: HY
authors.lastname: Chang
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Zijin
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Rong
authors.initials: R
authors.lastname: Fan
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Shaoxiong
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Diane
authors.initials: D
authors.lastname: Leland
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Liang
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Department of Pathology and Laboratory Medicine, Indiana University School of Medicine.
authors.forename: Jingmei
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D001706:Biopsy; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000175:diagnosis / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis*; D005557:Formaldehyde; D007150:Immunohistochemistry; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D016707:Tissue Fixation
chemical_list: D004279:DNA, Viral; D005557:Formaldehyde