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Detection of herpes simplex virus type 1 in herpetic ocular diseases by DNA‐DNA hybridization using a biotinylated DNA probe
Author(s) -
Nago Ryosuke,
Hayashi Kyoko,
Ochiai Hiroshi,
Kubota Yasuo,
Niwayama Seihachiro
Publication year - 1988
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890250303
Subject(s) - herpes simplex virus , virology , biology , biotinylation , hybridization probe , virus , microbiology and biotechnology , dna , herpesviridae , keratitis , dna–dna hybridization , viral disease , genetics
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.