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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2025-05-01
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| author | Abigail Dorothea Taylor Artur Trzebny Małgorzata Łośko Jerzy Franciszek Michalik Miroslawa Dabert |
| author_facet | Abigail Dorothea Taylor Artur Trzebny Małgorzata Łośko Jerzy Franciszek Michalik Miroslawa Dabert |
| author_sort | Abigail Dorothea Taylor |
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| description | 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. |
| format | Article |
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| issn | 2076-0817 |
| language | English |
| publishDate | 2025-05-01 |
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| series | Pathogens |
| spelling | doaj-art-97df09b38ca34fc4a3f20f40e1db88232025-08-20T01:56:44ZengMDPI AGPathogens2076-08172025-05-0114550610.3390/pathogens14050506A Novel <i>flaB</i> Gene-Based Profiling Approach for the Rapid and Accurate Detection of <i>Borreliella</i> and <i>Borrelia</i> Species in TicksAbigail Dorothea Taylor0Artur Trzebny1Małgorzata Łośko2Jerzy Franciszek Michalik3Miroslawa Dabert4Molecular Biology Techniques Laboratory, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, PolandMolecular Biology Techniques Laboratory, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, PolandMolecular Biology Techniques Laboratory, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, PolandDepartment of Animal Morphology, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, PolandMolecular Biology Techniques Laboratory, Faculty of Biology, Adam Mickiewicz University, 61-614 Poznan, PolandThe 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.https://www.mdpi.com/2076-0817/14/5/506Borreliaceaemolecular screeningflagellin<i>Borrelia burgdorferi</i> sensu latotargeted amplicon sequencing16S profiling |
| spellingShingle | Abigail Dorothea Taylor Artur Trzebny Małgorzata Łośko Jerzy Franciszek Michalik Miroslawa Dabert 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 Pathogens Borreliaceae molecular screening flagellin <i>Borrelia burgdorferi</i> sensu lato targeted amplicon sequencing 16S profiling |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | Borreliaceae molecular screening flagellin <i>Borrelia burgdorferi</i> sensu lato targeted amplicon sequencing 16S profiling |
| url | https://www.mdpi.com/2076-0817/14/5/506 |
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