Hybridization techniques provide improved sensitivity for HCMV detection and allow quantitation of the virus in clinical samples.
Hybridization techniques (slot-blot and in-situ hybridization assays) and immunostaining using murine monoclonal antibodies directed against different proteins of the human cytomegalovirus (HCMV) were compared for their sensitivity and specificity for detection of HCMV. A model system with HCMV infected human embryonic lung fibroblasts and lung biopsy specimens obtained from patients with culture positive HCMV interstitial pneumonia were used for evaluation of these techniques. The hybridization techniques were found to provide an improved sensitivity compared to immunostaining. Additionally a good correlation was found between the virus dose determined by TCID50 and the amount of viral DNA detected by slot-blot hybridization and by the number of autoradiographic silver grains per 100 cells per 2 weeks exposure time detected in the infected fibroblasts by in-situ hybridization. Thus, at least in the model system quantification of the virus was achieved by hybridization assays.
Einsele
H
H
Medizinische Klinik, Abteilung II, Universität Tübingen, F.R.G.
Vallbracht
A
A
Jahn
G
G
Kandolf
R
R
Müller
C A
CA
D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000453:epidemiology; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D005347:Fibroblasts / Q000032:analysis; D005860:Germany, West / Q000453:epidemiology; D006801:Humans; D007150:Immunohistochemistry; D009693:Nucleic Acid Hybridization; D011024:Pneumonia, Viral / Q000175:diagnosis / Q000453:epidemiology; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000276:immunology
D015342:DNA Probes; D004279:DNA, Viral; D014764:Viral Proteins