Related Experiment Video
Updated: Oct 26, 2025
07:21
Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing
Published on: August 25, 2018
13.1K
Data analysis workflow for the detection of canine vector-borne pathogens using 16 S rRNA Next-Generation Sequencing
Elton J R Vasconcelos1,2, Chayan Roy1, Joseph A Geiger1
1College of Veterinary Medicine, Western University of Health Sciences, 309 East 2nd Street, CA, 91766 - 1854, Pomona, USA.
BMC Veterinary Research
|August 1, 2021
Summary
Next-Generation Sequencing (NGS) offers a powerful new method for detecting multiple vector-borne pathogens (VBPs) in veterinary diagnostics. This approach enhances VBP detection beyond traditional PCR, improving public health surveillance.
Area of Science:
- Veterinary microbiology and diagnostics
- Next-Generation Sequencing (NGS) applications
- Bioinformatics and computational biology
Background:
- Vector-borne diseases (VBDs) pose significant challenges in human and veterinary medicine.
- Key bacterial VBPs include Anaplasma, Bartonella, Ehrlichia, and Spotted Fever Group Rickettsia.
- Current PCR-based diagnostics have limitations in detecting genetically diverse VBPs.
Purpose of the Study:
- To develop and present a novel methodology for genetic characterization of VBPs using NGS.
- To establish a comprehensive bioinformatics pipeline for VBP detection and analysis.
- To improve the diagnostic capabilities for vector-borne pathogens in veterinary medicine.
Main Methods:
- Utilized Next-Generation Sequencing (NGS) with broad-range 16S rRNA primers.
- Developed a Standard Operating Procedure (SOP) combining microbiome analysis tools and custom bioinformatic scripts.
- Validated the SOP using in vitro experiments with serial dilutions of plasmids representing key VBPs.
Main Results:
- Successfully detected single, double, and triple infections of target VBPs in canine DNA.
- Demonstrated the ability of NGS and the SOP to identify pathogens across different dilutions.
- Confirmed the capacity of the method for genetic characterization of multiple pathogens in a single sample.
Conclusions:
- NGS combined with advanced bioinformatics offers a significant improvement for VBP diagnostics.
- The presented workflow expands the molecular diagnostic toolbox for both veterinary and human medicine.
- This approach facilitates future applications of microbiome and metagenomic research in veterinary science.
