Routine diagnosis of herpes simplex virus (HSV) encephalitis by an internal DNA controlled HSV PCR and an IgG-capture assay for intrathecal synthesis of HSV antibodies.
BACKGROUND
The development of antiviral therapy increases the need for rapid, sensitive and reliable methods or combination of methods for diagnosis and monitoring herpes simplex encephalitis, HSE.


OBJECTIVE
Evaluation of diagnostic performance of three successively developed HSV PCR assays when combined with a new capture ELISA for HSV intrathecal antibody production (ITT).


METHODS
During a 3.6 year period a total of 4.206 CSF and serum samples from about 4.140 hospitalized patients with a tentative diagnosis of HSE were analyzed by a new ELISA for ITT. 1.962 CSF samples were examined also by PCR. Clinical signs and symptoms and additional tests were obtained on all ITT and/or PCR positive patients. In 1993 the PCR was a double PCR. In 1994 the PCR was a single PCR with internal inhibition control. Positive samples were confirmed by a different confirmative PCR to increase the specificity. From 1995 the PCR was as in 1994 but samples were no longer divided in the serology routine laboratory.


RESULTS
A total of 33 HSE cases was found (incidence 1.8 HSE per million people). All patients were treated with aciclovir. Three patients died, 9 patients had primary infection, 2 patients had HSE previously, and 2 patients relapsed. Only 11 patients recovered satisfactory. Of all 37 positive ITT 7 were unlikely positive. False positive PCR was seen in 1993 and 1994, due to sample-to-sample contamination during division of samples, but was not seen since 1995 when this procedure was changed. The test results depended on the state of the disease. Thus, the sensitivity, specificity, PPV and NPV for ITT were highest when performed more than 1 week after debut of symptoms whereas these values were highest using PCR within the first week.


CONCLUSIONS
Routine PCR diagnosis of HSE type 1 and 2 is a highly sensitive and specific method that should be performed together with serological ITT to cover the whole time span from debut of symptoms to several weeks after hospitalization.
Fomsgaard
A
A
Department of Virology, Statens Serum Institut, Copenhagen S, Denmark. afo@ssi.dk
Kirkby
N
N
Jensen
I P
IP
Vestergaard
B F
BF
D000212:Acyclovir / Q000627:therapeutic use; D000293:Adolescent; D000328:Adult; D000368:Aged; D000914:Antibodies, Viral / Q000097:blood / Q000134:cerebrospinal fluid; D000963:Antimetabolites / Q000627:therapeutic use; D000998:Antiviral Agents / Q000627:therapeutic use; D015139:Blotting, Southern; D002648:Child; D018792:Encephalitis, Viral / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000188:drug therapy; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006561:Herpes Simplex / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000188:drug therapy; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000134:cerebrospinal fluid; D007223:Infant; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D015995:Prevalence; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000914:Antibodies, Viral; D000963:Antimetabolites; D000998:Antiviral Agents; D007074:Immunoglobulin G; D000212:Acyclovir