Author | Pestana, Cristiane Pinheiro | |
Author | Lawson-Ferreira, Rafael | |
Author | Lessa-Aquino, Carolina | |
Author | Leal, Maria da Luz Fernandes | |
Author | Freire, Marcos da Silva | |
Author | Homma, Akira | |
Author | Medeiros, Marco Alberto | |
Access date | 2024-02-29T12:25:51Z | |
Available date | 2024-02-29T12:25:51Z | |
Document date | 2018 | |
Citation | PESTANA, Cristiane Pinheiro et al. Sanger-based sequencing technology for yellow fever vaccine genetic quality control. Journal of Virological Methods, v. 260, p. 82-87, Oct. 2018. | en_US |
ISSN | 0166-0934 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/62842 | |
Language | eng | en_US |
Publisher | Elsevier | en_US |
Rights | restricted access | en_US |
Title | Sanger-based sequencing technology for yellow fever vaccine genetic quality control | en_US |
Type | Article | en_US |
DOI | 10.1016/j.jviromet.2018.07.006 | |
Abstract | Yellow Fever (YF) is an acute viral hemorrhagic disease prevalent mainly in Africa and Americas, with 20–60% fatality rate in severe forms. Currently, antiviral drugs for the infection are not available, reinforcing the importance of vaccination in resident populations and travelers. Manufactured in 7 different countries, the YF vaccine was first created in 1937 and two substrains are used for production, 17DD and 17D-204. The vaccine produced in Bio-Manguinhos/Brazil uses 17DD substrain and more than 160 million doses have been exported to over 74 countries. The World Health Organization (WHO) recommends that new seed- and working-lots should have the viral genome sequenced in order to check vaccine genetic stability. The aim of this study was to develop and standardize a Sanger-based sequencing protocol for the genetic monitoring of the Brazilian 17DD vaccine. We designed 54 oligos to access the complete YF vaccine genome by RT-PCR and sequencing approach. After protocol standardization, we tested 45 vaccine lots and the corresponding secondary and working seed lots. All 45 lots presented 100% nucleotide identity to each other and to the seed lots. We also detected 2 heterogeneous positions at nucleotides 4523 (C/T) and 6673 (C/T) that may indicate a quasispecies diversity of YF 17DD strain. When compared to the Brazilian GenBank sequence YFU17066, the Brazilian 17DD vaccine presented 6 silent mutations. By applying the sequencing methodology to two YF 17D-204 strains, we showed that our method can also be used to sequence different YF vaccine virus. In summary, we have developed a robust method for the genetic monitoring of YF vaccines, which has been successfully applied in Bio-Manguinhos since 2009 and could also be used by other manufacturers for YF17D-based vaccines. There were no genetic variation in the Brazilian tested lots, highlighting the safety, production consistency and, more importantly, the genetic stability of BioManguinhos’ YF vaccine in the last 3 decades. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Rio de Janeiro, RJ, Brasil. | en_US |
Subject | Yellow Fever 17DD vaccine | en_US |
Subject | Genetic stability | en_US |
Subject | Quality control | en_US |
Embargo date | 2030-12-31 | |