A Multiplex Nanopore Sequencing Approach for the Detection of Multiple Arboviral Species.

Joilson Xavier,Vagner Fonseca,Talita Adelino,Felipe C M Iani,Glauco C Pereira,Myrian M Duarte,Mauricio Lima, Emerson Castro,Carla Oliveira,Hegger Fritsch,Natalia Guimarães,Ludmila O Lamounier,Fernanda Khouri Barreto, Camilo M M Braga de Oliveira,Crhistinne C Maymone Gonçalves, Danielle Malta Lima,Elaine C de Oliveira,Gislene G de Castro Lichs,Iago Gomes, Janaina Mazaro, Janete T N Rodrigues,Jayra Abrantes, Jeová K B Colares,Kleber G Luz,Luana Barbosa da Silva,Luiz Demarchi,Magaly C B Câmara,Marina C S Umaki Zardin, Rafaela Sabatini Mello Pinheiro, Rutilene Barbosa Souza, Simone K Haddad, Stephanni Figueiredo da Silva,Svetoslav N Slavov, Themis Rocha,Noelia Morel,Hector Chiparelli,Analía Burgueño,Victoria Bórmida,María N Cortinas, Rosario S Martín, Allan C Pereira, Marcelo F Dos Santos, Walter André Júnior,Jairo Mendez Rico,Leticia Franco,Alexander Rosewell,Rodrigo F do Carmo Said,Carlos F C de Albuquerque, Ethel L Noia Maciel, Marília Santini de Oliveira,Rivaldo Venâncio da Cunha, Livia C Vinhal Frutuoso,Ana M B de Filippis,Marta Giovanetti,Luiz Carlos Junior Alcantara

Viruses（2023）

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摘要

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.

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