A Multiplex Nanopore Sequencing Approach for the Detection of Multiple Arboviral Species.
Joilson XavierVagner FonsecaTalita AdelinoFelipe Campos de Melo IaniGlauco C PereiraMyrian M DuarteMaurício Teixeira LimaEmerson CastroCarla OliveiraHegger Machado FritschNatalia GuimarãesLudmila O LamounierFernanda Khouri BarretoCamilo M M Braga de OliveiraChristinne Cavalheiro Maymone GonçalvesDanielle Malta LimaElaine C de OliveiraGislene G de Castro LichsIago GomesJanaina MazaroJanete T N RodriguesJayra AbrantesJeová Keny Baima ColaresKleber G LuzLuana Barbosa da SilvaLuiz DemarchiMagaly C B CâmaraMarina C S Umaki ZardinRafaela Sabatini Mello PinheiroRutilene Barbosa SouzaSimone K HaddadStephanni Figueiredo da SilvaSvetoslav Nanev SlavovThemis RochaNoelia MorelHéctor ChiparelliAnalía BurgueñoVictoria BórmidaMaría N CortinasRosario S MartínAllan C PereiraMarcelo F Dos SantosWalter André JúniorJairo Mendez RicoLeticia FrancoAlexander RosewellRodrigo F do Carmo SaidCarlos Frederico Campelo de Albuquerque E MeloEthel L Noia MacielMarília Santini de OliveiraRivaldo Venâncio da CunhaLivia C Vinhal FrutuosoAna M B de FilippisMarta GiovanettiLuiz Carlos Junior AlcantaraPublished in: Viruses (2023)
The emergence and continued geographic expansion of arboviruses and the growing number of infected people have highlighted the need to develop and improve multiplex methods for rapid and specific detection of pathogens. Sequencing technologies are promising tools that can help in the laboratory diagnosis of conditions that share common symptoms, such as pathologies caused by emerging arboviruses. In this study, we integrated nanopore sequencing and the advantages of reverse transcription polymerase chain reaction (RT-PCR) to develop a multiplex RT-PCR protocol for the detection of Chikungunya virus (CHIKV) and several orthoflaviviruses (such as dengue ( Orthoflavivirus dengue ), Zika ( Orthoflavivirus zikaense ), yellow fever ( Orthoflavivirus flavi ), and West Nile ( Orthoflavivirus nilense ) viruses) in a single reaction, which provides data for sequence-based differentiation of arbovirus lineages.