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J Virol Methods ; 328: 114968, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38796133

ABSTRACT

Dengue fever, a mosquito-borne viral disease of significant public health concern in tropical and subtropical regions, is caused by any of the four serotypes of the dengue virus (DENV1-4). Cutting-edge technologies like next-generation sequencing (NGS) are revolutionizing virology, enabling in-depth exploration of DENV's genetic diversity. Here, we present an optimized workflow for full-genome sequencing of DENV 1-4 utilizing tiled amplicon multiplex PCR and Illumina sequencing. Our assay, sequenced on the Illumina MiSeq platform, demonstrates its ability to recover the full-length dengue genome across various viral abundances in clinical specimens with high-quality base coverage. This high quality underscores its suitability for precise examination of intra-host diversity, enriching our understanding of viral evolution and holding potential for improved diagnostic and intervention strategies in regions facing dengue outbreaks.


Subject(s)
Dengue Virus , Dengue , Genome, Viral , High-Throughput Nucleotide Sequencing , Multiplex Polymerase Chain Reaction , Serogroup , Whole Genome Sequencing , Dengue Virus/genetics , Dengue Virus/classification , Dengue Virus/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Dengue/virology , Dengue/diagnosis , Humans , Genome, Viral/genetics , Whole Genome Sequencing/methods , High-Throughput Nucleotide Sequencing/methods , RNA, Viral/genetics
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