A Novel <i>flaB</i> Gene-Based Profiling Approach for the Rapid and Accurate Detection of <i>Borreliella</i> and <i>Borrelia</i> Species in Ticks

A Novel <i>flaB</i> Gene-Based Profiling Approach for the Rapid and Accurate Detection of <i>Borreliella</i> and <i>Borrelia</i> Species in Ticks

The increasing incidence of tick-borne diseases in Europe necessitates the development of accurate and high-throughput molecular tools for detecting pathogens in tick populations. In this study, we present a novel <i>flaB</i> gene-based profiling method for the detection and identificati...

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Bibliographic Details
Main Authors: Abigail Dorothea Taylor, Artur Trzebny, Małgorzata Łośko, Jerzy Franciszek Michalik, Miroslawa Dabert
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Pathogens
Subjects:
Borreliaceae
molecular screening
flagellin
<i>Borrelia burgdorferi</i> sensu lato
targeted amplicon sequencing
16S profiling
Online Access:https://www.mdpi.com/2076-0817/14/5/506
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Summary:The increasing incidence of tick-borne diseases in Europe necessitates the development of accurate and high-throughput molecular tools for detecting pathogens in tick populations. In this study, we present a novel <i>flaB</i> gene-based profiling method for the detection and identification of <i>Borrelia</i> and <i>Borreliella</i> species in <i>Ixodes ricinus</i> ticks, combining newly designed primers with next-generation sequencing (NGS). The method was evaluated alongside conventional nested PCR targeting the <i>flaB</i> gene, as well as microbial profiling based on the V4 region of the <i>rrs</i> gene, using tick DNA extracted from 1088 specimens pooled into 94 samples. Our results demonstrate that the <i>flaB</i> gene-based profiling approach was the highest-performing out of the three methods, detecting Borreliaceae DNA in 83 DNA pools, compared to 58 and 56 pools using nested PCR and V4 <i>rrs</i> profiling, respectively. A total of 23 distinct <i>flaB</i> sequence variants were identified, corresponding to five Borreliaceae species: <i>Borreliella afzelii</i>, <i>Bl. garinii</i>, <i>Bl. valaisiana</i>, <i>Bl. burgdorferi</i>, and <i>Borrelia miyamotoi</i>. Additionally, the method enabled putative strain-level discrimination within species. Our results highlight the value of <i>flaB</i> gene-based profiling as a robust tool for ecological and epidemiological studies of Borreliaceae diversity in ticks.
ISSN:2076-0817

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