Detection of molluscum contagiosum virus DNA by in situ hybridization.
Three in situ hybridization protocols have been modified and used to comparatively examine formalin-fixed, paraffin-embedded tissue for the presence of molluscum contagiosum virus (MCV) DNA. Two cloned MCV 1 DNA fragments, MCV-1-B-D1 and MCV-1-B-F, were labelled with 32P-dCTP or digoxigenin-11-dUTP and used under conditions of high stringency to detect MCV DNA in sections of molluscum lesions. All methods were sensitive and were capable of detecting MCV after a minimum of 18 hours autoradiography (radiolabelled probes) or 3 to 18 hours (digoxigenin-labelled probes). However, one of the isotopic protocols produced less acceptable results in terms of non-specific background signals. A comparison of the DNA hybridization pattern with the underlying histopathological features of the molluscum sections revealed that there was not always a consistent association between the presence of detectable MCV DNA and cells containing the characteristic cytoplasmic inclusion bodies ("molluscum bodies") which are known to contain large numbers of virus particles. Although there is likely to be only limited scope for the application of this technique to diagnosis, it may prove useful for both retrospective and prospective epidemiological surveys. The in situ detection of MCV DNA may also be valuable for investigations into the cell biology of this imperfectly characterized virus.
Thompson
C H
CH
Department of Infectious Diseases, Faculty of Medicine, Sydney University, NSW.
Biggs
I M
IM
de Zwart-Steffe
R T
RT
D001345:Autoradiography; D015342:DNA Probes; D004279:DNA, Viral / Q000302:isolation & purification*; D004076:Digoxigenin; D006801:Humans; D007150:Immunohistochemistry; D008722:Methods; D008976:Molluscum Contagiosum / Q000382:microbiology / Q000473:pathology; D008977:Molluscum contagiosum virus / Q000235:genetics* / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D012867:Skin / Q000382:microbiology / Q000473:pathology
D015342:DNA Probes; D004279:DNA, Viral; D004076:Digoxigenin