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Journal of Clinical Microbiology, January 2002, p. 252-255, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.252-255.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Application of Real-Time Fluorescent PCR for Quantitative Assessment of Neospora caninum Infections in Organotypic Slice Cultures of Rat Central Nervous System Tissue

Norbert Müller,1* Nathalie Vonlaufen,1 Christian Gianinazzi,2 Stephen L. Leib,2 and Andrew Hemphill1*

Institute of Parasitology, University of Berne, Länggass-Strasse 122, 3012 Berne,1 Institute for Infectious Diseases, University of Berne, Friedbühlstrasse 51, 3010 Berne, Switzerland2

Received 14 May 2001/ Accepted 9 September 2001

The previously described Nc5-specific PCR test for the diagnosis of Neospora caninum infections was used to develop a quantitative PCR assay which allows the determination of infection intensities within different experimental and diagnostic sample groups. The quantitative PCR was performed by using a dual fluorescent hybridization probe system and the LightCycler Instrument for online detection of amplified DNA. This assay was successfully applied for demonstrating the parasite proliferation kinetics in organotypic slice cultures of rat brain which were infected in vitro with N. caninum tachyzoites. This PCR-based method of parasite quantitation with organotypic brain tissue samples can be regarded as a novel ex vivo approach for exploring different aspects of cerebral N. caninum infection.

* Corresponding author. Mailing address: Institute of Parasitology, Länggass-Str. 122, P.O. Box 8466, CH-3001 Berne, Switzerland. Phone: 41 31 631 2474. Fax: 41 31 631 2622. E-mail for Norbert Müller: nmueller{at}ipa.unibe.ch. E-mail for Andrew Hemphill: hemphill{at}ipa.unibe.ch.

Journal of Clinical Microbiology, January 2002, p. 252-255, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.252-255.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Vonlaufen, N., Guetg, N., Naguleswaran, A., Muller, N., Bjorkman, C., Schares, G., von Blumroeder, D., Ellis, J., Hemphill, A. (2004). In Vitro Induction of Neospora caninum Bradyzoites in Vero Cells Reveals Differential Antigen Expression, Localization, and Host-Cell Recognition of Tachyzoites and Bradyzoites. Infect. Immun. 72: 576-583 [Abstract] [Full Text]  


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