Real-time PCR for type-specific identification of herpes simplex in clinical samples: evaluation of type-specific results in the context of CNS diseases.
BACKGROUND
HSV-1 and HSV-2 cause CNS infections of dissimilar clinico-pathological characteristics with prognostic and therapeutic implications.


OBJECTIVE
To validate a type-specific real-time PCR that uses MGB/LNA Taqman probes and to review the virologico-clinical data of 25 eligible patients with non-neonatal CNS infections.


RESULTS
This real-time PCR was evaluated against conventional PCR (26 CSF and 20 quality controls), and LightCycler assay (51 mucocutaneous, 8 CSF and 32 quality controls) and culture/immunofluorescence (75 mucocutaneous) to assess typing with independent methods. Taqman real-time PCR detected 240 HSV genomes per ml CSF, a level appropriate for the management of patients, and provided unambiguous typing for the 104 positive (62 HSV-1 and 42 HSV-2) out the 160 independent clinical samples tested. HSV type diagnosed by Taqman real-time PCR predicted final diagnosis (meningitis versus encephalitis/meningoencephalitis, p<0.001) in 24/25 patients at time of presentation, in contrast to clinical evaluation.


CONCLUSIONS
Our real-time PCR, as a sensitive and specific means for type-specific HSV diagnosis, provided rapid prognostic information for patient management.
Meylan
Sylvain
S
Robert
Daniel
D
Estrade
Christine
C
Grimbuehler
Valérie
V
Péter
Olivier
O
Meylan
Pascal R
PR
Sahli
Roland
R
D001483:Base Sequence; D015342:DNA Probes; D018792:Encephalitis, Viral / Q000175:diagnosis* / Q000821:virology; D006561:Herpes Simplex / Q000175:diagnosis* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008587:Meningitis, Viral / Q000175:diagnosis* / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics
D015342:DNA Probes; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; C044525:glycoprotein B, herpes simplex virus type 2