Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood
The successful treatment of Lyme disease (LD) is contingent on accurate diagnosis. However, current laboratory detection assays lack sensitivity in the early stages of the disease. Because delayed diagnosis of LD incurs high healthcare costs and great suffering, new highly sensitive tests are in nee...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-03-01
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| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.651217/full |
| _version_ | 1819120941752385536 |
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| author | Jinyu Shan Ying Jia Louis Teulières Faizal Patel Martha R. J. Clokie |
| author_facet | Jinyu Shan Ying Jia Louis Teulières Faizal Patel Martha R. J. Clokie |
| author_sort | Jinyu Shan |
| collection | DOAJ |
| description | The successful treatment of Lyme disease (LD) is contingent on accurate diagnosis. However, current laboratory detection assays lack sensitivity in the early stages of the disease. Because delayed diagnosis of LD incurs high healthcare costs and great suffering, new highly sensitive tests are in need. To overcome these challenges, we developed an internally controlled quantitative PCR (Ter-qPCR) that targets the multicopy terminase large subunit (terL) gene encoded by prophages that are only found in LD-causing bacteria. The terL protein helps phages pack their DNA. Strikingly, the detection limit of the Ter-qPCR was analytically estimated to be 22 copies and one bacterial cell in bacteria spiked blood. Furthermore, significant quantitative differences was observed in terms of the amount of terL detected in healthy individuals and patients with either early or late disease. Together, the data suggests that the prophage-targeting PCR has significant power to improve success detection for LD. After rigorous clinical validation, this new test could deliver a step-change in the detection of LD. Prophage encoded markers are prevalent in many other pathogenic bacteria rendering this approach highly applicable to bacterial identification in general. |
| first_indexed | 2024-12-22T06:28:40Z |
| format | Article |
| id | doaj.art-a840ae55d52d42529a4db88c887708cc |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T06:28:40Z |
| publishDate | 2021-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-a840ae55d52d42529a4db88c887708cc2022-12-21T18:35:45ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-03-011210.3389/fmicb.2021.651217651217Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in BloodJinyu Shan0Ying Jia1Louis Teulières2Faizal Patel3Martha R. J. Clokie4Department of Genetics and Genome Biology, University of Leicester, Leicester, United KingdomDepartment of Genetics and Genome Biology, University of Leicester, Leicester, United KingdomPhelixRD Charity 230 Rue du Faubourg St Honoré, Paris, FranceDepartment of Genetics and Genome Biology, University of Leicester, Leicester, United KingdomDepartment of Genetics and Genome Biology, University of Leicester, Leicester, United KingdomThe successful treatment of Lyme disease (LD) is contingent on accurate diagnosis. However, current laboratory detection assays lack sensitivity in the early stages of the disease. Because delayed diagnosis of LD incurs high healthcare costs and great suffering, new highly sensitive tests are in need. To overcome these challenges, we developed an internally controlled quantitative PCR (Ter-qPCR) that targets the multicopy terminase large subunit (terL) gene encoded by prophages that are only found in LD-causing bacteria. The terL protein helps phages pack their DNA. Strikingly, the detection limit of the Ter-qPCR was analytically estimated to be 22 copies and one bacterial cell in bacteria spiked blood. Furthermore, significant quantitative differences was observed in terms of the amount of terL detected in healthy individuals and patients with either early or late disease. Together, the data suggests that the prophage-targeting PCR has significant power to improve success detection for LD. After rigorous clinical validation, this new test could deliver a step-change in the detection of LD. Prophage encoded markers are prevalent in many other pathogenic bacteria rendering this approach highly applicable to bacterial identification in general.https://www.frontiersin.org/articles/10.3389/fmicb.2021.651217/fullprophagesLyme diseasediagnosismulticopyBorreliaqPCR |
| spellingShingle | Jinyu Shan Ying Jia Louis Teulières Faizal Patel Martha R. J. Clokie Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood Frontiers in Microbiology prophages Lyme disease diagnosis multicopy Borrelia qPCR |
| title | Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood |
| title_full | Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood |
| title_fullStr | Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood |
| title_full_unstemmed | Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood |
| title_short | Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood |
| title_sort | targeting multicopy prophage genes for the increased detection of borrelia burgdorferi sensu lato s l the causative agents of lyme disease in blood |
| topic | prophages Lyme disease diagnosis multicopy Borrelia qPCR |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.651217/full |
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