Detection of cytomegalovirus by in situ hybridization using a digoxigenin-tailed oligonucleotide.
A non-isotopic in situ hybridization procedure was used to detect cytomegalovirus (CMV) sequences within routinely fixed tissue. A digoxigenin-tailed oligonucleotide was hybridized to sections of specimens obtained at autopsy from 2 patients with CMV infection. Hybrids were revealed by an alkaline phosphatase-conjugated anti-digoxigenin antibody. Serial sections were also assayed for the presence of CMV by in situ hybridization with a biotin-labelled cDNA probe and by immunohistochemistry and routinely stained for morphological evaluation. Results show that the two in situ hybridization procedures are equally sensitive but superior to the immunohistochemical detection of the viral antigen. Most cells positive for CMV DNA had the cytopathological features characteristic of CMV infection. A minor population of infected cells lacking morphological changes was also found. We recommend the routine application of the oligonucleotide-based assay because it is specific, easy and less expensive than other similar procedures.
Pacchioni
D
D
Department of Biomedical Sciences and Human Oncology, University of Turin School of Medicine, Italy.
Papotti
M
M
Bonino
F
F
Bussolati
G
G
Negro
F
F
D000469:Alkaline Phosphatase; D001483:Base Sequence; D003587:Cytomegalovirus / Q000737:chemistry*; D004247:DNA / Q000032:analysis; D004076:Digoxigenin / Q000737:chemistry*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000191:economics / Q000379:methods*; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry*
D015345:Oligonucleotide Probes; D004247:DNA; D000469:Alkaline Phosphatase; D004076:Digoxigenin