Rapid diagnosis of herpes simplex encephalitis by nested polymerase chain reaction assay of cerebrospinal fluid.
With the aim of improving early diagnosis of herpes simplex encephalitis a polymerase chain reaction (PCR) assay with two "nested" primer pairs was developed for the amplification of herpes simplex virus DNA in cerebrospinal fluid (CSF). Southern blotting was used to confirm the specificity of the amplification. The assay was applied to 151 CSF samples from 43 consecutive patients with herpes simplex encephalitis verified by the finding of herpes simplex virus/viral antigen in a brain biopsy sample or at necropsy (13) and/or intrathecal production of IgG antibody to the virus (40). As controls, 87 CSF samples from 60 patients with acute febrile focal encephalopathy (initially suspected to be herpes simplex encephalitis but excluded by the absence of intrathecal antibody synthesis) were tested. PCR detected herpes simplex virus DNA in 42 of the 43 patients with proven herpes simplex encephalitis; all but 1 were positive in the first CSF sample taken. The 1 PCR-negative patient had been treated with acyclovir from 20 h after the onset of symptoms. All the control subjects were PCR negative, as were 270 internal contamination controls. The PCR result remained positive in samples drawn up to 27 days after the onset of neurological symptoms. This method is a rapid and non-invasive means to diagnose herpes simplex encephalitis; it is highly sensitive and specific.
Aurelius
E
E
Department of Virology, Central Microbiological Laboratory, Stockholm, Sweden.
Johansson
B
B
Sköldenberg
B
B
Staland
A
A
Forsgren
M
M
D000212:Acyclovir / Q000627:therapeutic use; D000914:Antibodies, Viral / Q000134:cerebrospinal fluid; D000956:Antigens, Viral / Q000032:analysis; D001483:Base Sequence; D001923:Brain Chemistry; D004279:DNA, Viral / Q000134:cerebrospinal fluid*; D004660:Encephalitis / Q000134:cerebrospinal fluid* / Q000188:drug therapy / Q000209:etiology; D005069:Evaluation Studies as Topic; D006561:Herpes Simplex / Q000134:cerebrospinal fluid* / Q000188:drug therapy; D006801:Humans; D007074:Immunoglobulin G / Q000134:cerebrospinal fluid; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018139:Simplexvirus / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D007074:Immunoglobulin G; D000212:Acyclovir