Luminometric microplate hybridization for detection of varicella-zoster virus PCR product from cerebrospinal fluid.
We modified and optimized a new microplate hybridization assay to detect the varciella-zoster virus (VZV) PCR product, and studied cerebrospinal fluid (CSF) samples of 287 patients with meningitis, encephalitis or other neurological diseases or symptoms. Specific antibodies to VZV and reference antigens were determined by enzyme immunoassay from serum and CSF, they were then compared with clinical findings and with the results obtained by VZV-PCR using different detection methods for VZV-specific amplified DNA. VZV DNA was found in the CSF of 25 patients using the microplate hybridization assay and chemiluminescence detection for amplified DNA. All 25 CSF samples were also positive in Southern blotting. Among the patients, 10 had chickenpox, 4 had shingles, and 11 had no rash at all. The detection rate of VZV-specific DNA by microplate hybridization was 30% higher than that obtained by conventional agarose gel electrophoresis. In most patients the diagnosis was confirmed by demonstrating specific intrathecal antibody production to VZV but not to other viruses. These results indicate the presence of VZV in the central nervous system (CNS) in many patients with chickenpox or shingles, and even in patients without a rash. The microplate hybridization assay based on chemiluminescence detection improves considerably the detection rate of the VZV-PCR product compared to agarose gel electrophoresis and will add to the list of recognized VZV infections in the CNS. It is especially useful in cases where there is no cutaneous manifestation.
Koskiniemi
M
M
Haartman Institute, Department of Virology, University of Helsinki, Finland. marjaleena.Koskiniemi@helsinki.fi
Mannonen
L
L
Kallio
A
A
Vaheri
A
A
D002644:Chickenpox / Q000134:cerebrospinal fluid / Q000821:virology*; D002675:Child, Preschool; D004279:DNA, Viral / Q000134:cerebrospinal fluid*; D018792:Encephalitis, Viral / Q000134:cerebrospinal fluid / Q000821:virology; D005260:Female; D006562:Herpes Zoster / Q000134:cerebrospinal fluid / Q000821:virology*; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D008163:Luminescent Measurements; D008297:Male; D008587:Meningitis, Viral / Q000134:cerebrospinal fluid / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*
D004279:DNA, Viral