Oligonucleotide probe for herpes virus: use in paraffin sections.
A method is described for in situ hybridization detection and typing of herpes simplex virus (HSV) using alkaline phosphatase-labeled synthetic oligonucleotide probes in paraffin tissue sections. Sections mounted on slides are prehybridized and denatured before the probe mixture is added. Hybridization proceeds for 1 h at 60 degrees C. Detection of the alkaline phosphatase label is performed using a nitro blue tetrazolium and 5-bromo-4-chloro-3-indolyl phosphate substrate. Specific hybridization with HSV type 1 DNA was found in sections of herpetic esophagitis and encephalitis. There was no discernible background staining. Hybridization with an oligonucleotide probe specific for HSV type 2 was negative. No hybridization occurred to sections of cytomegalovirus- or adenovirus-infected tissue. The development of this technique expands the utility of synthetic oligonucleotide probes to include hybridization reactions in routinely processed and paraffin-embedded tissue. The use of directly labeled oligonucleotide probes for tissue in situ hybridization overcomes problems of probe contamination with vector plasmid DNA, nonspecific avidin binding to tissue, and the danger and inconvenience of working with radioactive materials.
Bruner
J M
JM
Department of Pathology, University of Texas M.D. Anderson Cancer Center, Houston.
D000469:Alkaline Phosphatase; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D006561:Herpes Simplex / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010232:Paraffin
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D010232:Paraffin; D000469:Alkaline Phosphatase