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Title: The detection of viral antigen in the cerebrospinal fluid of patients with herpes simplex virus encephalitis
Author: McLean, B. N.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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A highly sensitive capture sandwich antibody assay was developed for the detection of Herpes Simplex Virus alkaline nuclease in cerebrospinal fluid. Chemiluminescence, using Isoluminol labelled antibody and p-Iodophenol enhanced Horse-Radish Peroxidase catalysed Luminol, was compared with Horse-Radish Peroxidase colorimetric detection in the assay. Luminol was superior to Isoluminol based chemiluminescence, but this was no more sensitive than colorimetric detection. In vitro the final optimised colorimetric assay had a sensitivity of 20 plaque forming units of Herpes Simplex Virus. A method for dissociation of immune complexed alkaline nuclease, using Hydrochloric acid as a chaotropic agent, was optimised for use with the assay. The assay was applied to 12 cerebrospinal fluid samples from 9 cases of Herpes Simplex Virus Encephalitis. All cases conformed to accepted criteria for the diagnosis of the disease in the absence of brain biopsy. All the tested samples were positive for alkaline nuclease, which was present in either free or immune complexed form or both. The earliest positive sample was day 1 of the clinical illness, and samples were positive up to day 56, which was the longest interval from onset of symptoms to lumbar puncture in these patients. Free alkaline nuclease was present in the early stages of the disease, being replaced by immune complexed alkaline nuclease after day 13, a time when locally synthesised anti-HSV1 IgG was also detected. A total of 35 control patients were also studied. Five out of 6 patients with suspected Herpes Simplex Virus Encephalitis or myelitis, and one case of definite herpes myelitis were positive. Two of the remaining 28 cases were positive: both of these were patients with Subacute Sclerosing Panencephalitis. No demyelinating, inflammatory, other infectious or non-neurological disorders were positive by the assay.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (M.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available