Polymerase chain reaction and viral culture techniques to detect HSV in small volumes of cerebrospinal fluid; an experimental mouse encephalitis study.
A technique is described for the detection of HSV-DNA in very small volumes (5-10 microliters) of cerebrospinal fluid (CSF). The method was evaluated in CSF samples of 4-6-week-old mice inoculated with HSV-1 via the corneal route. The sensitivity of the PCR assay was compared with results of spin-amplified viral culture with immunofluorescent visualization (SAC/IF), routine viral culture (RVC) and radioactive dot-blot hybridization (DBA) in CSF samples obtained from other mice. The results show the PCR to be superior over the other techniques: infectious virus or viral DNA in CSF was demonstrated by PCR, SAC/IF, RVC and DBA in 68, 55, 20 and 2.5%, respectively. These results show the feasibility of the PCR for a rapid, non-invasive diagnosis of human HSV-encephalitis.
Boerman
R H
RH
Department of Neurology, University of Leiden, The Netherlands.
Arnoldus
E P
EP
Raap
A K
AK
Bloem
B R
BR
Verhey
M
M
van Gemert
G
G
Peters
A C
AC
van der Ploeg
M
M
D000818:Animals; D004279:DNA, Viral / Q000134:cerebrospinal fluid*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004660:Encephalitis / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D005455:Fluorescent Antibody Technique; D005784:Gene Amplification; D006561:Herpes Simplex / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D006801:Humans; D015151:Immunoblotting; D008297:Male; D051379:Mice; D018139:Simplexvirus / Q000235:genetics* / Q000302:isolation & purification
D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase