Previous Article | Next Article 
Journal of Clinical Microbiology, July 2000, p. 2638-2642, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Detection of Herpes Simplex Virus DNA by
Real-Time PCR
Harald H.
Kessler,1,*
Gerhard
Mühlbauer,2
Beate
Rinner,1
Evelyn
Stelzl,1
Annemarie
Berger,3
Hans-Wilhelm
Dörr,3
Brigitte
Santner,1
Egon
Marth,1 and
Holger
Rabenau3
Institute of Hygiene,
Karl-Franzens-University Graz, A-8010 Graz,1 and
Roche Diagnostics GmbH, A-1211 Vienna,2
Austria, and Institute of Medical Virology,
Johann-Wolfgang-Goethe-University Frankfurt, D-60596 Frankfurt am
Main, Germany3
Received 27 January 2000/Returned for modification 3 April
2000/Accepted 8 May 2000
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
104 copies per ml, i.e., 12.5 copies per run. When samples
of the First European Union Concerted Action HSV Proficiency Panel were tested, 2 × 103 to 5 × 103 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.
*
Corresponding author. Mailing address: Molecular
Diagnostics Laboratory, Institute of Hygiene, KF-University Graz,
Universitaetsplatz 4, A-8010 Graz, Austria. Phone: 43(316)380-4363.
Fax: 43(316)380-9649. E-mail:
harald.kessler{at}kfunigraz.ac.at.
Journal of Clinical Microbiology, July 2000, p. 2638-2642, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Reil, H., Bartlime, A., Drerup, J., Grewing, T., Korn, K.
(2008). Clinical Validation of a New Triplex Real-Time Polymerase Chain Reaction Assay for the Detection and Discrimination of Herpes simplex Virus Types 1 and 2. J. Mol. Diagn.
10: 361-367
[Abstract]
[Full Text]
-
Allawi, H. T., Li, H., Sander, T., Aslanukov, A., Lyamichev, V. I., Blackman, A., Elagin, S., Tang, Y.-W.
(2006). Invader plus method detects herpes simplex virus in cerebrospinal fluid and simultaneously differentiates types 1 and 2.. J. Clin. Microbiol.
44: 3443-3447
[Abstract]
[Full Text]
-
Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F.
(2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev.
19: 165-256
[Abstract]
[Full Text]
-
Namvar, L., Olofsson, S., Bergstrom, T., Lindh, M.
(2005). Detection and Typing of Herpes Simplex Virus (HSV) in Mucocutaneous Samples by TaqMan PCR Targeting a gB Segment Homologous for HSV Types 1 and 2. J. Clin. Microbiol.
43: 2058-2064
[Abstract]
[Full Text]
-
Enomoto, Y., Yoshikawa, T., Ihira, M., Akimoto, S., Miyake, F., Usui, C., Suga, S., Suzuki, K., Kawana, T., Nishiyama, Y., Asano, Y.
(2005). Rapid Diagnosis of Herpes Simplex Virus Infection by a Loop-Mediated Isothermal Amplification Method. J. Clin. Microbiol.
43: 951-955
[Abstract]
[Full Text]
-
Ndjoyi-Mbiguino, A., Ozouaki, F., Legoff, J., Mbopi-Keou, F.-X., Si-Mohamed, A., Onas, I. N., Avoune, E., Belec, L.
(2003). Comparison of Washing and Swabbing Procedures for Collecting Genital Fluids To Assess Cervicovaginal Shedding of Herpes Simplex Virus Type 2 DNA. J. Clin. Microbiol.
41: 2662-2664
[Abstract]
[Full Text]
-
Anderson, T. P., Werno, A. M., Beynon, K. A., Murdoch, D. R.
(2003). Failure To Genotype Herpes Simplex Virus by Real-Time PCR Assay and Melting Curve Analysis Due to Sequence Variation within Probe Binding Sites. J. Clin. Microbiol.
41: 2135-2137
[Abstract]
[Full Text]
-
Weidmann, M., Meyer-Konig, U., Hufert, F. T.
(2003). Rapid Detection of Herpes Simplex Virus and Varicella-Zoster Virus Infections by Real-Time PCR. J. Clin. Microbiol.
41: 1565-1568
[Abstract]
[Full Text]
-
Barratt, K., Mackay, J. F.
(2002). Improving Real-Time PCR Genotyping Assays by Asymmetric Amplification. J. Clin. Microbiol.
40: 1571-1572
[Full Text]
-
Mackay, I. M., Arden, K. E., Nitsche, A.
(2002). Real-time PCR in virology. Nucleic Acids Res
30: 1292-1305
[Abstract]
[Full Text]
-
Garcia, S., Crance, J. M., Billecocq, A., Peinnequin, A., Jouan, A., Bouloy, M., Garin, D.
(2001). Quantitative Real-Time PCR Detection of Rift Valley Fever Virus and Its Application to Evaluation of Antiviral Compounds. J. Clin. Microbiol.
39: 4456-4461
[Abstract]
[Full Text]
-
Verstrepen, W. A., Kuhn, S., Kockx, M. M., Van De Vyvere, M. E., Mertens, A. H.
(2001). Rapid Detection of Enterovirus RNA in Cerebrospinal Fluid Specimens with a Novel Single-Tube Real-Time Reverse Transcription-PCR Assay. J. Clin. Microbiol.
39: 4093-4096
[Abstract]
[Full Text]
-
Read, S. J., Mitchell, J. L., Fink, C. G.
(2001). LightCycler Multiplex PCR for the Laboratory Diagnosis of Common Viral Infections of the Central Nervous System. J. Clin. Microbiol.
39: 3056-3059
[Abstract]
[Full Text]
-
Kessler, H. H., Muhlbauer, G., Stelzl, E., Daghofer, E., Santner, B. I., Marth, E.
(2001). Fully Automated Nucleic Acid Extraction: MagNA Pure LC. Clin. Chem.
47: 1124-1126
[Full Text]