Ingrid Haas, Gerhard Mühlbauer, Michael Bozic, Evelyn Stelzl, Christoph Koidl, Annemarie Berger, Jörg Berg, Adriana Vince, Holger Rabenau, Harald H. Kessler
Evaluation of a new assay for detection of herpes simplex virus type 1 and type 2 DNA by real-time PCR/Evaluierung eines neuen Tests zur Detektierung von Herpes simplex Virus Typ 1 und Typ 2 DNA mittels Real-time PCR
Molecular detection of herpes simplex virus (HSV) DNA is recognized as reference standard assay for the sensitive and specific diagnosis of central nervous system and genital infections caused by HSV. In this study, a qualitative molecular assay based on automated DNA extraction on the MagNA Pure LC (Roche Applied Science, Mannheim, Germany) and a commercially available kit, the LightCycler – HSV 1/2 Detection Kit (Roche), were evaluated. This kit includes a homologous internal control. The accuracy and the detection limit of the new molecular assay were determined with samples from European Union Concerted Action HSV Proficiency Panels. A total of 153 clinical specimens including cerebrospinal fluids, genital swabs, and oral swabs were investigated and the results were compared to those obtained from a home-brew molecular assay based on manual DNA extraction and real-time polymerase chain reaction (PCR). When the accuracy of the new molecular assay was tested, all positive samples except for the one containing 3.0 × 102–9.0 × 102 HSV type 1 (HSV-1) genome equivalents (GE) per ml could be detected. All negative samples tested negative. When the detection limit was determined, 6.0 × 102–1.8 × 103 HSV-1 GE per ml, i.e. 12 to 36 GE per LightCycler (LC) capillary and 2.0 × 102–6.0 × 102 HSV type 2 (HSV-2) GE per ml, i.e. 4 to 12 GE per LC capillary could consistently be detected. Results obtained from clinical specimens corresponded to 100% to those obtained by the molecular assay based on manual DNA extraction and real-time PCR. In seven clinical samples, an unexpected melting peak was detected. In conclusion, the new molecular assay allows rapid detection of HSV-1 and HSV-2 DNA in the routine diagnostic laboratory. The inclusion of a homologous internal control ensures accurate interpretation of negative results.
LaboratoriumsMedizin, Walter de Gruyter
Print ISSN: 0025-8466
Volume: 28, 08/2004
Pages: 361 - 367
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