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

2023; Multidisciplinary Digital Publishing Institute; Volume: 16; Issue: 1 Linguagem: Inglês

10.3390/v16010023

1999-4915

Joilson Xavier, Vagner Fonseca, Talita Adelino, Felipe Campos de Melo Iani, Glauco Carvalho Pereira, Myrian Morato Duarte, Maurício Lima, Emerson Castro, Carla de Oliveira, Hegger Fritsch, Natália Rocha Guimarães, Ludmila Oliveira Lamounier, Fernanda Khouri Barreto, Camilo M. M. Braga de Oliveira, Crhistinne Cavalheiro Maymone Gonçalves, Danielle Malta Lima, Elaine Cristina de Oliveira, Gislene Garcia de Castro Lichs, Iago Pachêco Gomes, Janaína Mazaro, Janete T. N. Rodrigues, Jayra Abrantes, Jeová Keny Baima Colares, Kléber Giovanni Luz, Luana Barbosa da Silva, Luiz Henrique Ferraz Demarchi, Magaly C. B. Câmara, Marina Castilhos Souza Umaki Zardin, R.S. Pinheiro, Rutilene Barbosa Souza, Simone Kashima, Stephanni Figueiredo da Silva, Svetoslav Nanev Slavov, Themis Rocha, Noelia Morel, Héctor Chiparelli, Analía Burgueño, Victoria Bórmida, María Noel Cortinas, Rosario S. Martín, A. Cascaes Pereira, Marcelo Ferreira dos Santos, Walter André Júnior, Jairo Méndez‐Rico, Leticia Franco, Alexander Rosewell, Rodrigo Fabiano do Carmo Said, Carlos F. Campelo de Albuquerque, Ethel L. Noia Maciel, Marília Santini de Oliveira, Rivaldo Venâncio da Cunha, Lívia Carla Vinhal Frutuoso, Ana María Bispo de Filippis, Marta Giovanetti, Luíz Carlos Júnior Alcântara,

Phytoplasmas and Hemiptera pathogens

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.