Detection of herpes simplex virus type 1 in herpetic ocular diseases by DNA-DNA hybridization using a biotinylated DNA probe.
A diagnostic hybridization assay for detecting herpes simplex virus type 1 (HSV-1) in ocular specimens was developed using cloned viral DNA as a probe. This hybridization assay is based on visualizing a biotinylated probe that is hybridized to the target DNA by a streptavidin/alkaline phosphatase system. The time required for performing this assay system is only two days. This assay system could detect a probe which had been hybridized to as little as 1 pg of homologous DNA and did not cross-react with DNA of other human herpes viruses except that of herpes simplex virus type 2 (HSV-2) which showed weak cross-reactivity. The assay system was applied to experimental keratitis in albino rabbits and clinical specimens. In experimental keratitis in rabbits it was possible to detect HSV-1 DNA in the eye swab samples at least until the ninth day after virus inoculation. Five clinical specimens collected from patients with corneal ulcer or blepharitis contained HSV-1 DNA in spite of the failure of demonstration of viral antigen and/or virus isolation in two cases.
Nago
R
R
Department of Ophthalmology, Toyama Medical and Pharmaceutical University, Japan.
Hayashi
K
K
Ochiai
H
H
Kubota
Y
Y
Niwayama
S
S
D000818:Animals; D002460:Cell Line; D003315:Cornea / Q000382:microbiology; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D004195:Disease Models, Animal; D006801:Humans; D007635:Keratitis, Dendritic / Q000382:microbiology*; D009693:Nucleic Acid Hybridization; D011817:Rabbits; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*
D004279:DNA, Viral