Detection of cytomegalovirus DNA using probes labelled with digoxigenin

Abstract

Cloned HindIII restriction fragments of cytomegalovirus (CMV) DNA (strain AD169) were labelled with biotin, digoxigenin or 32P and used as probes to detect CMV DNA. Probes biotinylated by nick translation, random hexanucleotide priming (RHP) or "photobiotin" were able to detect 10-50 pg of homologous DNA. Probes labelled with digoxigenin or 32P by RHP detected 0.1 pg of homologous DNA, and 1-10 CMV-infected fibroblasts. Comparison of digoxigenin- and 32P-labelled probes in a DNA hybridisation assay on 186 urine specimens demonstrated that these probes were of similar sensitivity, detecting CMV DNA in 40 and 41 specimens, respectively. Positive results were obtained using this hybridisation assay with 11 of 14 specimens (79%) yielding CMV by virus isolation, and with 35 other specimens obtained from patients with laboratory evidence of CMV infection or symptoms compatible with CMV disease. Thus digoxigenin-labelled probes may provide an assay that can detect CMV DNA in specimens yielding a negative result by virus isolation.