Evaluation of a commercial microbial enrichment kit used prior DNA extraction to improve the molecular detection of vector-borne pathogens from naturally infected dogs

Oney, Kristina; Koo, Melody; Roy, Chayan; Ren, Shuo; Qurollo, Barbara A.; Juhasz, Nicholas B.; Vasconcelos, Elton J R; Oakley, Brian B.; Diniz, Pedro

doi:10.1016/j.mimet.2021.106163

Cited by 4 publications

(1 citation statement)

References 46 publications

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“…Molecular biological methods include nucleic acid hybridization, gene recombination, PCR technology, etc, which are suitable for detection of uncommon or new pathogenic microorganisms. These methods are rapid, sensitive and speci c, but it requires a high level of experimental design or complicated operational procedures [13,[18][19][20][21]. Metabolomics is a high-resolution analytical method that obtains detailed information on various metabolites produced in biological systems.…”

Section: Introductionmentioning

confidence: 99%

Fluorescent Probe combined with Novel Rapid Microbial Analyzer for Detection of Contaminated Pathogen in Coconut

Cui

Zhong

Deng

et al. 2022

Preprint

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Food safety is facing great challenges for preventing foodborne diseases caused by pathogenic pollution, especially in resource-limited area. The rapid detection technique of microorganisms, such as immunological method and molecular biological method, plays a crucial key in the timely bioanalysis and disease treatment strategies. However, it is difficult for these methods to simultaneously meet the criteria of simple operation, high specificity and sensitivity, as well as low cost. Herein, we introduced a Rapid Microbial Analyzer (RMA) that can provide a strong signal with high signal-to-noise ratio via embodying the metabolic features of microbial enzyme target and selective fluorescent substrate. The technique is confirmed high accuracy for the detection of E. coli in the coconut water with a limit of detection of 1 CFU/mL, and could severe as an acute pathogenic detection tool. This device provides a one-step strategy for bacterial detection and is a great leap in the development of easy to operate instrumentation, and could be a great aid for global pathogenic monitoring.

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Section: Introductionmentioning

confidence: 99%

Fluorescent Probe combined with Novel Rapid Microbial Analyzer for Detection of Contaminated Pathogen in Coconut

Cui

Zhong

Deng

et al. 2022

Preprint

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Blood Supplementation Enhances Bartonella henselae Growth and Molecular Detection of Bacterial DNA in Liquid Culture

Liedig,

Neupane,

Lashnits

et al. 2023

Microbiol Spectr

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This study aims to improve diagnostic detection of Bartonella henselae . Patient samples are combined with enriched bacterial cultures aimed at growing Bartonella henselae for the best possible chance at detection. However, current Bartonella growth methods could be improved. The DNA extraction method used by most laboratories should also be optimized. Sheep blood was added to increase the growth of Bartonella henselae and multiple DNA extraction methods were to be compared to each other.

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Data analysis workflow for the detection of canine vector-borne pathogens using 16 S rRNA Next-Generation Sequencing

et al. 2021

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Background Vector-borne diseases (VBDs) impact both human and veterinary medicine and pose special public health challenges. The main bacterial vector-borne pathogens (VBPs) of importance in veterinary medicine include Anaplasma spp., Bartonella spp., Ehrlichia spp., and Spotted Fever Group Rickettsia. Taxon-targeted PCR assays are the current gold standard for VBP diagnostics but limitations on the detection of genetically diverse organisms support a novel approach for broader detection of VBPs. We present a methodology for genetic characterization of VBPs using Next-Generation Sequencing (NGS) and computational approaches. A major advantage of NGS is the ability to detect multiple organisms present in the same clinical sample in an unsupervised (i.e. non-targeted) and semi-quantitative way. The Standard Operating Procedure (SOP) presented here combines industry-standard microbiome analysis tools with our ad-hoc bioinformatic scripts to form a complete analysis pipeline accessible to veterinary scientists and freely available for download and use at https://github.com/eltonjrv/microbiome.westernu/tree/SOP. Results We tested and validated our SOP by mimicking single, double, and triple infections in genomic canine DNA using serial dilutions of plasmids containing the entire 16 S rRNA gene sequence of (A) phagocytophilum, (B) v. berkhoffii, and E. canis. NGS with broad-range 16 S rRNA primers followed by our bioinformatics SOP was capable of detecting these pathogens in biological replicates of different dilutions. These results illustrate the ability of NGS to detect and genetically characterize multi-infections with different amounts of pathogens in a single sample. Conclusions Bloodborne microbiomics & metagenomics approaches may help expand the molecular diagnostic toolbox in veterinary and human medicine. In this paper, we present both in vitro and in silico detailed protocols that can be combined into a single workflow that may provide a significant improvement in VBP diagnostics and also facilitate future applications of microbiome research in veterinary medicine.

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Evaluation of a commercial microbial enrichment kit used prior DNA extraction to improve the molecular detection of vector-borne pathogens from naturally infected dogs

Cited by 4 publications

References 46 publications

Fluorescent Probe combined with Novel Rapid Microbial Analyzer for Detection of Contaminated Pathogen in Coconut

Fluorescent Probe combined with Novel Rapid Microbial Analyzer for Detection of Contaminated Pathogen in Coconut

Blood Supplementation Enhances Bartonella henselae Growth and Molecular Detection of Bacterial DNA in Liquid Culture

Data analysis workflow for the detection of canine vector-borne pathogens using 16 S rRNA Next-Generation Sequencing

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