This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kessler, H H Articles by Marth, E Search for Related Content PubMed PubMed Citation Articles by Kessler, H H Articles by Marth, E

Detection of herpes simplex virus DNA from cerebrospinal fluid by PCR and a rapid, nonradioactive hybridization technique.

Institute of Hygiene, KF-University Graz, Austria.

ABSTRACT

A molecular assay for the detection of herpes simplex virus (HSV), including a novel, nonradioactive hybridization technique, was evaluated with a total of 123 cerebrospinal fluid specimens. After DNA extraction, specific HSV DNA sequences were amplified with digoxigenin-labeled primers derived from the DNA polymerase gene-coding region from HSV. Amplified products were detected by the Enzymun-Test DNA detection assay (Boehringer, Mannheim, Federal Republic of Germany), which uses biotinylated probes. Amplification with nonlabeled primers and then Southern blotting and nonradioactive detection of hybrids by the digoxigenin technique was the reference system. The sensitivities of the molecular assays were determined with 10-fold dilutions of plasmid pS4 with the SalI restriction fragment of the DNA polymerase gene obtained from the HSV type 1 strain Angelotti. The Enzymun assay was able to detect all of the 16 positive samples, giving 100% agreement with the Southern blot hybridization results. Optical density values were widely separated for the positive and negative groups of specimens. Ten copies of plasmid pS4 per microliter could be distinctly detected by the Enzymun assay. The cutoff was determined for the hybridization assay, and an equivocal zone was defined. The whole molecular assay including the Enzymun-Test DNA detection proved to be sensitive and easy to use. It may contribute to the rapid and safe detection of HSV DNA in cerebrospinal fluid.

This article has been cited by other articles:

• Kessler, H. H., Mühlbauer, G., Rinner, B., Stelzl, E., Berger, A., Dörr, H.-W., Santner, B., Marth, E., Rabenau, H. (2000). Detection of Herpes Simplex Virus DNA by Real-Time PCR. J. Clin. Microbiol. 38: 2638-2642 [Abstract] [Full Text]  
• Hofgartner, W. T., Huhmer, A. F.R., Landers, J. P., Kant, J. A. (1999). Rapid Diagnosis of Herpes Simplex Encephalitis Using Microchip Electrophoresis of PCR Products. Clin. Chem. 45: 2120-2128 [Abstract] [Full Text]  
• Tang, Y.-W., Mitchell, P. S., Espy, M. J., Smith, T. F., Persing, D. H. (1999). Molecular Diagnosis of Herpes Simplex Virus Infections in the Central Nervous System. J. Clin. Microbiol. 37: 2127-2136 [Full Text]  
• Domingues, R. B., Lakeman, F. D., Mayo, M. S., Whitley, R. J. (1998). Application of Competitive PCR to Cerebrospinal Fluid Samples from Patients with Herpes Simplex Encephalitis. J. Clin. Microbiol. 36: 2229-2234 [Abstract] [Full Text]