Journal
TICKS AND TICK-BORNE DISEASES
Volume 4, Issue 5, Pages 409-416Publisher
ELSEVIER GMBH
DOI: 10.1016/j.ttbdis.2013.04.004
Keywords
PCR/nested PCR; Hard ticks; Borrelia burgdorferi sensu lato complex; Anaplasma phagocytophilum; Babesia spp.
Categories
Ask authors/readers for more resources
Most epidemiological studies on tick-borne pathogens employ PCR approaches. Prevalences determined by this method particularly depend on the efficiency and sensitivity of the applied protocol, which can be influenced by various factors. In the current study, we examined crucial points to improve PCR strategies with respect to sensitivity as well as specificity and developed optimized and easily practicable PCR/nested PCR approaches for the detection of different tick-borne pathogens. Among them were Ana plasma phagocytophilum, Borrelia burgdorferi sensu lato complex, and Babesia spp., for which the novel protocol was successfully applied to perform species determination by direct sequencing of PCR amplicons. The occurrence of double and multiple infections of different species of the same genus were detected by chromatogram analyses exhibiting mixed sequences. Furthermore, we suggest employing a standard plasmid containing the appropriate target sequence followed by verification of the encountered detection limit with an addition of tick DNA spiked with the appropriate pathogen DNA. Approaches standardized in this manner would facilitate the comparison of prevalence studies. (C) 2013 Elsevier GmbH. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available