TY - JOUR
T1 - Use of chimeric DNA-RNA primers in quantitative PCR for detection of Ehrlichia canis and Babesia canis
AU - Peleg, Ofer
AU - Baneth, Gad
AU - Eyal, Osnat
AU - Inbar, Jacob
AU - Harrus, Shimon
PY - 2009/10
Y1 - 2009/10
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=70349420939&partnerID=8YFLogxK
U2 - 10.1128/AEM.00720-09
DO - 10.1128/AEM.00720-09
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C2 - 19633128
AN - SCOPUS:70349420939
SN - 0099-2240
VL - 75
SP - 6396
EP - 6398
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
IS - 19
ER -