A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats

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• dc.contributor.author Albó Timor, Andreu
• dc.contributor.author Lucati, Federica
• dc.contributor.author Bartumeus, Frederic
• dc.contributor.author Caner, Jenny
• dc.contributor.author Escartin, Santi
• dc.contributor.author Mariani, Simone
• dc.contributor.author Palmer, John R. B.
• dc.contributor.author Ventura, Marc
• dc.date.accessioned 2024-04-22T06:18:24Z
• dc.date.available 2024-04-22T06:18:24Z
• dc.date.issued 2023
• dc.description.abstract Background. Determination of the interactions between hematophagous mosquitoes and their human hosts is of great importance for better understanding the transmission dynamics of mosquito-borne arboviruses and developing effective strategies to mitigate risk. Genetic analysis of human and mosquito DNA can play a key role in this, but commercial kits for human short tandem repeat (STR) genotyping are expensive and do not allow for the simultaneous STR analysis of host and vector DNA. Here, we present an inexpensive and straightforward STR-loci multiplex system capable of simultaneously amplifying Aedes albopictus and human STRs from blood-fed mosquitoes. Additionally, we examine the effect of storage methods and post-feeding time on the integrity of host DNA. Methods. Thirty-five STRs (16 human and 19 Ae. albopictus STRs) subdivided in three multiplexes were tested for amplification and scoring reliability. Under laboratory conditions we compared the efficacy of two preservation methods (absolute ethanol vs lysis buffer) on the integrity of host DNA in Ae. albopictus blood meals. We also evaluated the effect of post-feeding time by sacrificing blood-fed mosquitoes at different time intervals after feeding, and we assessed our ability to detect multiple feedings. To determine if the system can be employed successfully under field conditions, we carried out a preliminary study using field-collected Ae. albopictus. Results. All 35 STRs amplified consistently in the laboratory. Lysis buffer performed better than absolute ethanol in terms of allele peak height and clarity of electropherograms. Complete human DNA profiles could be obtained up to 48 h following the blood meal. Analysis of multiple feedings confirmed that peak heights can be used as a proxy to determine post-feeding time and thus derive the number of different people bitten by a mosquito. In the field trial, amplification was successful for 32 STRs. We found human DNA signal in 38 of the 61 field-collected mosquitoes (62%), of which 34 (89%) had ingested a single blood meal, while four (11%) contained double meals. Conclusions. Our new genotyping system allows fast and reliable screening of both host and vector species, and can be further adapted to other mosquito species living in close contact with humans.
• dc.description.sponsorship This work was supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (Grant agreement No. 853271).
• dc.format.mimetype application/pdf
• dc.identifier.citation Albó A, Lucati F, Bartumeus F, Caner J, Escartin S, Mariani S, et al. A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats. Parasites Vectors. 2023 Oct 5;16(1):347. DOI: 10.1186/s13071-023-05977-w
• dc.identifier.doi http://dx.doi.org/10.1186/s13071-023-05977-w
• dc.identifier.issn 1756-3305
• dc.identifier.uri http://hdl.handle.net/10230/59848
• dc.language.iso eng
• dc.publisher BioMed Central
• dc.relation.ispartof Parasites Vectors. 2023 Oct 5;16(1):347
• dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/853271
• dc.rights This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. he Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Aedes albopictus
• dc.subject.keyword Blood meal analysis
• dc.subject.keyword Human fingerprinting
• dc.subject.keyword STR genotyping
• dc.subject.keyword Vector-host interaction
• dc.title A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion