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Development of a Standardized Sanger-Based Method for Partial Sequencing and Genotyping of Dengue Viruses

Authors :
Fabiola Cruz-López
Jorge L. Muñoz-Jordán
Gilberto A. Santiago
Glenda L. González
Source :
Journal of Clinical Microbiology. 57
Publication Year :
2019
Publisher :
American Society for Microbiology, 2019.

Abstract

The global expansion of dengue viruses (DENV-1 to DENV-4) has contributed to the divergence, transmission, and establishment of genetic lineages of epidemiological concern; however, tracking the phylogenetic relationships of these virus is not always possible due to the inability of standardized sequencing procedures in resource-limited public health laboratories. Consequently, public genomic data banks contain inadequate representation of geographical regions and historical periods. In order to improve detection of the DENV-1 to DENV-4 lineages, we report the development of a serotype-specific Sanger-based method standardized to sequence DENV-1 to DENV-4 directly from clinical samples using universal primers that detect most DENV genotypes. The resulting envelope protein coding sequences are analyzed for genotyping with phylogenetic methods. We evaluated the performance of this method by detecting, amplifying, and sequencing 54 contemporary DENV isolates, including 29 clinical samples, representing a variety of genotypes of epidemiological importance and global presence. All specimens were sequenced successfully and phylogenetic reconstructions resulted in the expected genotype classification. To further improve genomic surveillance in regions where dengue is endemic, this method was transferred to 16 public health laboratories in 13 Latin American countries, to date. Our objective is to provide an accessible method that facilitates the integration of genomics with dengue surveillance.

Details

ISSN :
1098660X and 00951137
Volume :
57
Database :
OpenAIRE
Journal :
Journal of Clinical Microbiology
Accession number :
edsair.doi.dedup.....011e6eb5ac752955f0c8b513b6a199c1
Full Text :
https://doi.org/10.1128/jcm.01957-18