Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Kessler, HH; Mühlbauer, G; Rinner, B; Stelzl, E; Berger, A; Dörr, HW; Santner, B; Marth, E; Rabenau, H.
Detection of Herpes simplex virus DNA by real-time PCR.
J Clin Microbiol. 2000; 38(7):2638-2642
Doi: 10.1128/JCM.38.7.2638-2642.2000
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- Führende Autor*innen der Med Uni Graz
- Kessler Harald
- Co-Autor*innen der Med Uni Graz
- Marth Egon
- Rinner Beate
- Santner Brigitte
- Stelzl Evelyn
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- Abstract:
- Molecular detection of herpes simplex virus (HSV) DNA is recognized as the reference standard assay method for the sensitive and specific diagnosis of central nervous system infections caused by HSV. In this study, a molecular assay based on real-time PCR on the LightCycler (LC) instrument was evaluated and compared with a home-brew molecular assay. The detection limit of the LC assay was determined with 10-fold dilutions of plasmid pS4 with the SalI restriction fragment of the DNA polymerase gene and with the First European Union Concerted Action HSV Proficiency Panel. A total of 59 cerebrospinal fluid (CSF) specimens were investigated for the comparative study. With plasmid pS4, the detection limit of the LC assay was found to be 10(4) copies per ml, i.e., 12.5 copies per run. When samples of the First European Union Concerted Action HSV Proficiency Panel were tested, 2x10(3) to 5x10(3) HSV type 1 genome equivalents (GE) per ml, i.e., 2.5 to 6.3 GE per run, could consistently be detected. There was a correlation between the LC assay and the home-brew assay in 55 of 59 specimens. In conclusion, the LC assay allows very rapid detection of HSV DNA in CSF. It was found to be laborsaving and showed sufficient sensitivity.
- Find related publications in this database (using NLM MeSH Indexing)
- Adult -
- Central Nervous System Viral Diseases - virology
- Cerebrospinal Fluid - virology
- DNA, Viral - cerebrospinal fluid
- Female -
- Herpes Simplex - virology
- Herpesvirus 1, Human - isolation and purification
- Herpesvirus 2, Human - isolation and purification
- Humans -
- Male -
- Middle Aged -
- Polymerase Chain Reaction - instrumentation Polymerase Chain Reaction - methods