Detection of Cytomegalovirus‐infected cells in autopsy material by in situ hybridization

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Abstract
A non-radioactive in situ hybridization (ISH) method was elaborated to detect cytomegalovirus (CMV)-infected cells in tissue specimens processed for diagnostic routine histopathology. A biotinylated CMV-DNA probe was hybridized following a) four different enzymatic predigestions, b) progressively increasing denaturation periods, and then detected by c) streptavidin-biotin, d) a monoclonal antibody against biotin using a three-stage alkaline phosphatase anti-alkaline phosphatase (APAAP)-technique, and e) combining c + d. Autopsy specimens obtained from an infant with acquired CMV-infection, six patients with AIDS, five patients clinically and serologically without CMV-infection, and preoperative needle core biopsies from six renal allografts served as material. ISH was specific and more sensitive when compared to immunohistochemical (IMH) detection of CMV-antigens by a monoclonal antibody. ISH was concluded to be a rapid, practical, and sensitive tool in daily diagnostic histopathology.