SOP

SOP standardizes detection and genetic characterization of canine vector-borne pathogens using 16S rRNA next-generation sequencing.

Key Features:

• Unsupervised Detection: Employs a non-targeted 16S rRNA NGS approach to detect multiple pathogens simultaneously in a single sample.
• Semi-Quantitative Analysis: Provides semi-quantitative estimates of pathogen presence based on sequencing read abundances.
• Broad Detection Capability: Uses broad-range 16S rRNA primers to detect bacterial veterinary pathogens including Anaplasma spp., Bartonella spp., Ehrlichia spp., and Spotted Fever Group Rickettsia.
• Validation through Simulation: Validated by simulating single, double, and triple infections via serial dilutions of plasmids containing entire 16S rRNA gene sequences of A. phagocytophilum, B. v. berkhoffii, and E. canis spiked into genomic canine DNA.
• Integration with Industry-Standard Tools: Combines established microbiome analysis tools with custom bioinformatic scripts for identification and genetic characterization of vector-borne pathogens.

Scientific Applications:

• Veterinary Diagnostics: Enhances molecular detection and genetic characterization of canine vector-borne bacterial pathogens.
• Public Health Research: Contributes to public health efforts by improving detection and characterization of VBPs that may also affect humans.
• Microbiome Research: Enables investigation of bloodborne microbial communities in veterinary microbiome studies.

Methodology:

Sequencing of the 16S rRNA gene by next-generation sequencing followed by processing of resulting data with established microbiome analysis tools and custom bioinformatic scripts to identify and characterize vector-borne pathogens.