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Updated: Jul 30, 2025
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Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR
Published on: February 4, 2018
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Development of a Syndromic Molecular Diagnostic Assay for Tick-Borne Pathogens Using Barcoded Magnetic Bead
Nazleeen Mohseni1, Mariann Chang1, Kathryn Garcia1
1College of Veterinary Medicine, Western University of Health Sciences, Pomona, California, USA.
Microbiology Spectrum
|May 11, 2023
Summary
This study introduces a new diagnostic method for detecting tick-borne diseases like Anaplasma phagocytophilum and Borrelia burgdorferi. The barcoded magnetic bead technology offers a sensitive and rapid way to identify multiple pathogens simultaneously from one sample.
Area of Science:
- Infectious Disease Diagnostics
- Molecular Biology
- Biotechnology
Background:
- Current serological tests for infectious diseases are often costly, slow, and lack sensitivity and specificity.
- Diagnosing tick-borne diseases is challenging due to low pathogen loads in samples and the absence of sensitive multiplex diagnostic methods.
- This limits clinical testing and epidemiological surveillance of tick-borne infections.
Purpose of the Study:
- To demonstrate the principle of a novel multiplex diagnostic assay for simultaneous detection of tick-borne pathogens.
- To develop a sensitive, specific, and rapid diagnostic approach for Anaplasma phagocytophilum and Borrelia burgdorferi.
- To establish a foundation for a cost-effective multiplex diagnostic platform for various tick-borne diseases.
Main Methods:
- Utilized barcoded magnetic bead technology on the BioCode 2500 system for simultaneous pathogen detection.
- Employed PCR amplification with specific primers targeting conserved genes of Anaplasma phagocytophilum and Borrelia burgdorferi.
- Developed a hybridization assay using biotinylated PCR products and specific probes on magnetic beads, detected via a high-quantum-yield fluorophore.
Main Results:
- Achieved a limit of detection (LOD) of 2.81 50% tissue culture infection dose (TCID50)/mL for A. phagocytophilum and 1 CFU/mL for B. burgdorferi.
- Demonstrated an LOD of 1.8 genome copies/reaction for both pathogens, with a high signal-to-background ratio.
- Showcased 100% positive and negative agreement in performance evaluations using patient and spiked samples, with no observed cross-reactivity.
Conclusions:
- The developed assay is a proof-of-principle for a sensitive and specific multiplex diagnostic approach for tick-borne pathogens.
- This technology enables simultaneous detection of multiple pathogens from a single sample with a short turnaround time.
- The assay can be upgraded for broader clinical application, aiding early diagnosis, treatment, and public health surveillance.
