Use of chimeric DNA-RNA primers in quantitative PCR for detection of Ehrlichia canis and Babesia canis

Ofer Peleg*, Gad Baneth, Osnat Eyal, Jacob Inbar, Shimon Harrus

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

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.

Original languageAmerican English
Pages (from-to)6396-6398
Number of pages3
JournalApplied and Environmental Microbiology
Volume75
Issue number19
DOIs
StatePublished - Oct 2009

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