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Detection of Rickettsia in ticks using loop-mediated isothermal amplification (LAMP)

2025-03-06Journal article Open access
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dc.contributor.authorLansdell, Samantha
dc.contributor.authorHassan, Marwa M.
dc.contributor.authorLa Ragione, Roberto
dc.contributor.authorBetson, Martha
dc.contributor.authorNuncio, MS
dc.contributor.authorLopes de Carvalho, Isabel
dc.contributor.authorZé-Zé, Líbia
dc.contributor.authorde Sousa, Rita
dc.contributor.authorCutler, Sally
dc.contributor.editorDavis, Joshua S.
dc.date.accessioned2025-11-12T11:26:44Z
dc.date.available2025-11-12T11:26:44Z
dc.date.issued2025-03-06
dc.description.abstractObjectives: The objective of the study was to develop a Loop-mediated Isothermal Amplification (LAMP) assay for screening of Rickettsia species circulating in ticks using the citrate synthase gene (gltA). The LAMP assay employed portable visualisation methods, making the assay more field-suitable. Furthermore, prior methods have not used gltA as the target, despite proven success in Polymerase Chain Reaction (PCR) methods. Methods: Using an alignment of 72 DNA sequences (comprised of 21 Rickettsia species) from GenBank we designed a novel set of gltA LAMP primers. Evaluation used DNA from 12 Rickettsia species as positive controls (extracted from cultures or naturally infected ticks) alongside a panel of negative controls representing different bacterial species. Subsequently this assay was used to screen 295 Ixodes ricinus and 24 I. hexagonus ticks collected from the UK (including northern and southern England and northern Scotland). Results: LAMP successfully detected 11 out of 12 (91.7%) Rickettsia species, excluding Rickettsia akari. Among 319 ticks collected in the UK, three were positive for Rickettsia (0.9%). All three positives were I. ricinus ticks, while none of the 24 I. hexagonus ticks were positive. Results were confirmed using a published PCR method. Sanger sequencing of PCR amplicons generated for each positive tick showed that they were all R. helvetica. Conclusions: This study introduces a novel field-applicable LAMP protocol for efficient Rickettsia screening in ticks to better assess its prevalence and consequent health risks. Furthermore, this assay has proven suitability for rickettsial detection in I. ricinus ticks, which has been reported as unsuccessful in previous European studies.eng
dc.description.sponsorshipEuropean Society of Clinical Microbiology and Infectious Diseases
dc.identifier.citationCMI Communications. 2025 jun;2(2):105069. doi:10.1016/j.cmicom.2025.105069. Epub 2025 Mar 6
dc.identifier.doihttps://doi.org/10.1016/j.cmicom.2025.105069
dc.identifier.issn2950-5909
dc.identifier.urihttp://hdl.handle.net/10400.18/10596
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier
dc.relation.hasversionhttps://www.sciencedirect.com/science/article/pii/S2950590925000113?ref=pdf_download&fr=RR-2&rr=9564673adabbe3b9
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectRickettsia
dc.subjectLAMP
dc.subjectTick
dc.subjectIxodes ricinus
dc.subjectField
dc.subjectInfecções Sistémicas e Zoonoses
dc.titleDetection of Rickettsia in ticks using loop-mediated isothermal amplification (LAMP)eng
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue2
oaire.citation.startPage105069
oaire.citation.titleCMI Communications
oaire.citation.volume2
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyNameNúncio
person.familyNameLopes de Carvalho
person.familyNameMarques Zé-Zé
person.givenNameMaria Sofia
person.givenNameIsabel
person.givenNameLíbia Maria
person.identifierZ-1027-2018
person.identifierU-5730-2017
person.identifier.ciencia-idC715-3703-B77D
person.identifier.ciencia-id5E12-F1E2-CAE3
person.identifier.ciencia-id6F16-5659-41DB
person.identifier.orcid0000-0001-5182-6150
person.identifier.orcid0000-0002-4693-3952
person.identifier.orcid0000-0001-7258-1439
person.identifier.scopus-author-id1911430
relation.isAuthorOfPublicatione4e97d6a-e2ff-4078-bcdd-f31706151251
relation.isAuthorOfPublication694de188-6bdf-4129-82ac-7f4e00edfad7
relation.isAuthorOfPublicationcf9eaec1-8966-42a6-a2ae-ca379cd73508
relation.isAuthorOfPublication.latestForDiscoverye4e97d6a-e2ff-4078-bcdd-f31706151251

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