ArticleParasites & vectors2013
A system to simultaneously detect tick-borne pathogens based on the variability of the 16S ribosomal genes.
Article in Parasites & vectors, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Anaplasmosis in the Amazon: diagnostic challenges, persistence, and control ofFrontiers in veterinary science · 2025Review
- Impact of twoPhysiology and molecular biology of plants : an international journal of functional plant biology · 2024Article
- Tick-Borne Pathogens Screening Using a Multiplex Real-Time Polymerase Chain Reaction-Based Method.Acta parasitologica · 2023Article
- Diverse plant promoting bacterial species differentially improve tomato plant fitness under water stress.Frontiers in plant science · 2023Article
- Article
- Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder.Scientific reports · 2021Article
- Comparative proteomics of the vector Dermacentor reticulatus revealed differentially regulated proteins associated with pathogen transmission in response to laboratory infection with Rickettsia slovaca.Parasites & vectors · 2019Article
- Recent strategies for the diagnosis of early Lyme disease.Science progress · 2018Review
- The natural infection of birds and ticks feeding on birds with Rickettsia spp. and Coxiella burnetii in Slovakia.Experimental & applied acarology · 2016Article
- Rapid Identification and Characterization of Francisella by Molecular Biology and Other Techniques.The open microbiology journal · 2016Article
- Rickettsia species in fleas collected from small mammals in Slovakia.Parasitology research · 2015Article
- Inhibition of the endosymbiont "Candidatus Midichloria mitochondrii" during 16S rRNA gene profiling reveals potential pathogens in Ixodes ticks from Australia.Parasites & vectors · 2015Article
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3 authors.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundDNA microarrays can be used to quickly and sensitively identify several different pathogens in one step. Our previously developed DNA microarray, based on the detection of variable regions in the 16S rDNA gene (rrs), which are specific for each selected bacterial genus, allowed the concurrent detection of Borrelia spp., Anaplasma spp., Francisella spp., Rickettsia spp. and Coxiella spp.
methodsIn this study, we developed a comprehensive detection system consisting of a second generation DNA microarray and quantitative PCRs. New oligonucleotide capture probes specific for Borrelia burgdorferi s.l. genospecies and Candidatus Neoehrlichia mikurensis were included. This new DNA microarray system required substantial changes in solution composition, hybridization conditions and post-hybridization washes.
resultsThis second generation chip displayed high specificity and sensitivity. The specificity of the capture probes was tested by hybridizing the DNA microarrays with Cy5-labeled, PCR-generated amplicons encoding the rrs genes of both target and non-target bacteria. The detection limit was determined to be 10(3) genome copies, which corresponds to 1-2 pg of DNA. A given sample was evaluated as positive if its mean fluorescence was at least 10% of the mean fluorescence of a positive control. Those samples with fluorescence close to the threshold were further analyzed using quantitative PCRs, developed to identify Francisella spp., Rickettsia spp. and Coxiella spp. Like the DNA microarray, the qPCRs were based on the genus specific variable regions of the rrs gene. No unspecific cross-reactions were detected. The detection limit for Francisella spp. was determined to be only 1 genome copy, for Coxiella spp. 10 copies, and for Rickettsia spp., 100 copies.
conclusionsOur detection system offers a rapid method for the comprehensive identification of tick-borne bacteria, which is applicable to clinical samples. It can also be used to identify both pathogenic and endosymbiontic bacteria in ticks for eco-epidemiological studies, tick laboratory colony testing, and many other applications.
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