Direct detection of Molluscum contagiosum virus in clinical specimens by in situ hybridization using biotinylated probe.
Molluscum contagiosum virus (MCV) is an unclassified poxvirus which has recently become recognized as causing a major sexually transmitted disease. At present no assay is available for specific detection of MCV because the virus cannot be serially propagated in cell culture. Since MCV produces an abortive, limited growth with some cytopathic effect in certain cell lines, we were able to develop an in situ hybridization assay for detection of MCV genome in clinical specimens. Human fetal diploid lung cell monolayers were infected with clinical specimens, and after proper incubation and fixation in paraformaldehyde, hybridization was performed under full stringency conditions with a molecularly cloned biotinylated probe. Only MCV infected cells showed homology to the MCV probe with a purple-brown cytoplasmic staining. Additionally, we have described an in situ hybridization assay for direct detection of MCV genome in formalin-fixed, paraffin-embedded biopsies. Characteristic intracytoplasmic Molluscum bodies (Henderson-Paterson bodies) were detected in stratum spinosum cells of the epidermis. Striking staining similarities have been observed between in situ hybridization and haematoxylin-eosin cytostaining. These procedures are the first successful identification of MCV genome in clinical samples by molecular hybridization, with sensitivity and specificity equal to or greater than electron microscopy.
Forghani
B
B
Viral and Rickettsial Disease Laboratory, California State Department of Health Services, Berkeley.
Oshiro
L S
LS
Chan
C S
CS
Hurst
J W
JW
Dennis
J
J
Darai
G
G
Warford
A L
AL
Cohen
R M
RM
D015139:Blotting, Southern; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D006801:Humans; D008977:Molluscum contagiosum virus / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016612:Paraffin Embedding; D013997:Time Factors
D015342:DNA Probes; D004247:DNA