Journal
ANALYTICA CHIMICA ACTA
Volume 1001, Issue -, Pages 106-111Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2017.10.037
Keywords
DNA biosensor; Quartz crystal microbalance (QCM); DNA-DNA hybridization; Ehrlichia canis
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Funding
- Thai Research fund via Royal Golden Jubilee Ph.D. Program [PHD/0139/2552]
- Department of Biochemistry, Faculty of Medicine
- Graduate School of Srinakharinwirot University
- Department of Physical Chemistry, Faculty of Chemistry, University of Vienna
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Ehrlichia canis is an intracellular parasitic bacterium and arthropod-borne pathogen that receives growing attention, because it leads to increasing morbidity and mortality in animals. It does so by causing canine monocytotropic ehrlichiosis (CME). Infected canines may lack obvious clinical signs and stay in chronic stage. Herein we report a rapid screening method based on PCR assay combined with quartz crystal microbalance (QCM) to design a DNA sensor for detecting E. canis in early stages of infection. The test relies on DNA amplification of target nucleotide sequences via PCR followed by detecting DNA-DNA hybridization using QCM. The approach did not result in any cross-hybridization toward other blood bacteria or parasites in dogs, such as Anaplasma platys, Babesia canis and Trypanosoma spp, but turned out selective for the target species. The limit of detection of QCM was as low as 4.1 x 10(9) molecules/mu l of 289 bp E. canis PCR products corresponding to 22 copy numbers/mu l of E. canis. Furthermore, the technique is also simple, does not require complicated equipment and can in principle be reused. (C) 2017 Elsevier B.V. All rights reserved.
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