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Title: QCM-based rapid detection of PCR amplification products of Ehrlichia canis
Authors: Bunroddith K.
Viseshakul N.
Chansiri K.
Lieberzeit P.
Keywords: Bacteria
DNA sequences
Quartz crystal microbalances
Cross hybridization
DNA amplification
DNA biosensors
DNA-DNA hybridization
Ehrlichia canis
Limit of detection
Nucleotide sequences
PCR amplification
Polymerase chain reaction
bacterium detection
blood sampling
controlled study
copy number variation
cross hybridization
DNA hybridization
Ehrlichia canis
gene amplification
limit of detection
molecular probe
nucleotide sequence
polymerase chain reaction
priority journal
process optimization
quartz crystal microbalance
screening test
dog disease
Ehrlichia canis
equipment design
isolation and purification
polymerase chain reaction
quartz crystal microbalance
bacterial DNA
DNA, Bacterial
Dog Diseases
Ehrlichia canis
Equipment Design
Limit of Detection
Polymerase Chain Reaction
Quartz Crystal Microbalance Techniques
Issue Date: 2018
Abstract: 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 × 109 molecules/μl of 289 bp E. canis PCR products corresponding to 22 copy numbers/μl of E. canis. Furthermore, the technique is also simple, does not require complicated equipment and can in principle be reused. © 2017 Elsevier B.V.
ISSN: 32670
Appears in Collections:SCOPUS 1983-2021

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