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Applied and Environmental Microbiology, October 2009, p. 6393-6398, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00720-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Use of Chimeric DNA-RNA Primers in Quantitative PCR for Detection of Ehrlichia canis and Babesia canis{triangledown} ,{dagger}

Ofer Peleg,1* Gad Baneth,2 Osnat Eyal,2 Jacob Inbar,1 and Shimon Harrus2

Genaphora Ltd., 4 Habarzel Street, Tel Aviv 69710, Israel,1 Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel2

Received 28 March 2009/ Accepted 18 July 2009

To overcome the problem of nonspecific by-products in quantitative PCR (qPCR) assays, we constructed DNA-RNA chimeric primers and evaluated their use in the detection and quantification of the Ehrlichia canis 16S rRNA, Babesia canis Hsp70, and canine β-actin genes. Several RNA bases were incorporated into specific positions in the DNA primers, while no RNA stretches were allowed. qPCR reactions were carried out without preamplification steps. This resulted in decreased formation of undesirable by-products and a 10-fold increase in assay sensitivity.

* Corresponding author. Mailing address: Genaphora Ltd., 4 Habarzel St., Tel Aviv 69710, Israel. Phone: 972-544525922. Fax: 972-36496664. E-mail: ofer.peleg{at}gmail.com. Address for reprint requests: Shimon Harrus, Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel. E-mail: harrus{at}agri.huji.ac.il

{triangledown} Published ahead of print on 24 July 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, October 2009, p. 6393-6398, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00720-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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