A rapid simple in situ hybridization method for herpes simplex virus employing a synthetic biotin‐labeled oligonucleotide probe: A comparison with immunohistochemical methods for HSV detection

Abstract

We examined 28 paraffin-embedded tissue specimens with histologic evidence of herpes virus infection by in situ hybridization (ISH) utilizing manual capillary action technology (MicroProbe Staining System) and a 21 base synthetic multibiotinylated oligonucleotide probe from the HSV glycoprotein C region. The results were compared to a rapid simple immunohistochemical (IHC) protocol for detection of HSV proteins. HSV was detected by ISH and IHC in all but one specimen which was shown to be positive for varicella zoster virus by direct fluorescent antibody studies. Hybridization signal was confined to the nucleus in all cases. Staining was identified in cells with early as well as late cytopathic effect. IHC produced intense nuclear and/or cytoplasmic signal in infected cells and stained in areas of necrosis which were otherwise spared by ISH. HSV was detected by IHC and/or ISH in 3/5 specimens with histology suggestive of, but not diagnostic for, HSV infection. Both techniques were sensitive and specific for HSV, resulted in rapid detection of the pathogen in routinely processed tissues, and may be useful in cases where the histologic impression is equivocal for HSV infection. ISH for HSV may be preferred because it can identify early HSV infection, which in turn can be treated with antiviral agents.

Immunohistology of Infectious Diseases

“…However, the diagnosis of HSV infection can be difficult when the characteristic intranuclear inclusions or multinucleated cells, or both, are absent or when the amount of tissue in a biopsy specimen is small. 29,30 Although polyclonal antibodies against the major HSV glycoprotein antigens are sensitive, they do not allow distinction between HSV-1 and HSV-2 because these two viruses are antigenically similar. 27,28 Immunohistochemistry using either polyclonal or monoclonal antibodies against HSV antigens has proven to be a sensitive and specific technique to diagnose HSV infections ( Fig.…”

Interaction of VEGF to gastric low grade MALT lymphoma by Helicobacter heilmannii infection in C57/BL/6 mice

“…Some of the tissues were embedded in paraffi n and 4-µm sections were created; in situ hybridization using a microprobe method (Wang, 1994) was then carried out with the reported specifi c primers (Hhe1, 2, 3, 4, 5) (Trebesius, 2001). Vol.…”

“ Candidatus Helicobacter heilmannii” from a Cynomolgus Monkey Induces Gastric Mucosa-Associated Lymphoid Tissue Lymphomas in C57BL/6 Mice

“…Ultrathin sections were then cut, counterstained with uranyl acetate, and observed with a JEOL 1200 EX-II electron microscope at an accelerating voltage of 80 kV. Some of the tissues were embedded in paraffin, and 4-m sections were cut; in situ hybridization using a microprobe method (30) was then carried out with previously described specific primers (Hhe1, Hhe2, Hhe3, Hhe4, and Hhe5) (29). Sense probes were used in the control experiment.…”