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用于所有登革病毒血清型包膜基因扩增和测序的新型引物组的开发

The Development of New Primer Sets for the Amplification and Sequencing of the Envelope Gene of All Dengue Virus Serotypes.

作者信息

Fraenkel Stefania, Nabeshima Takeshi, Xayavong Dalouny, Nguyen Thi Thanh Ngan, Xu Qiang, Kapandji Merveille, Yamao Kano, Balingit Jean Claude, Pandey Basu Dev, Morita Kouichi, Hasebe Futoshi, Ngwe Tun Mya Myat, Takamatsu Yuki

机构信息

Department of Virology, Institute of Tropical Medicine (ITM-NU), Nagasaki University, Nagasaki 852-8523, Japan.

School of Tropical Medicine and Global Health (TMGH), Nagasaki University, Nagasaki 852-8523, Japan.

出版信息

Microorganisms. 2024 May 28;12(6):1092. doi: 10.3390/microorganisms12061092.

DOI:10.3390/microorganisms12061092
PMID:38930474
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11205395/
Abstract

Dengue virus (DENV) poses a significant threat to global health, infecting approximately 390 million people annually. This virus comprises four serotypes capable of causing severe disease. Genetic analyses are crucial for understanding the epidemiology, evolution, and spread of DENV. Although previous studies have focused on the envelope protein-coding (E) gene, only a few primers can efficiently detect and amplify the viral genes from multiple endemic countries simultaneously. In this study, we designed degenerate primer pairs for each DENV serotype to amplify and sequence the entire E gene, using globally representative sequences for each serotype. These primers were validated using DENV isolates from various Asian countries and demonstrated broad-spectrum detection capabilities and high-quality sequences. The primers provide effective tools for genetic analysis in the regions affected by dengue, aiding strain identification and epidemiological studies during outbreaks.

摘要

登革病毒（DENV）对全球健康构成重大威胁，每年感染约3.9亿人。这种病毒由四种血清型组成，能够引发严重疾病。基因分析对于了解登革病毒的流行病学、进化和传播至关重要。尽管先前的研究集中在包膜蛋白编码（E）基因上，但只有少数引物能够同时有效地从多个流行国家检测和扩增病毒基因。在本研究中，我们针对每种登革病毒血清型设计了简并引物对，以扩增和测序整个E基因，并使用每种血清型的全球代表性序列。这些引物通过来自亚洲不同国家的登革病毒分离株进行了验证，显示出广谱检测能力和高质量序列。这些引物为登革热影响地区的基因分析提供了有效的工具，有助于在疫情爆发期间进行毒株鉴定和流行病学研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/780f287645c4/microorganisms-12-01092-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/078ca55c2e33/microorganisms-12-01092-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/fa48edba4888/microorganisms-12-01092-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/b197b4d69581/microorganisms-12-01092-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/780f287645c4/microorganisms-12-01092-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/078ca55c2e33/microorganisms-12-01092-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/fa48edba4888/microorganisms-12-01092-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/b197b4d69581/microorganisms-12-01092-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2fe4/11205395/780f287645c4/microorganisms-12-01092-g004.jpg

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本文引用的文献

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J Med Virol. 2023 Nov;95(11):e29255. doi: 10.1002/jmv.29255.
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Next-generation diagnostic test for dengue virus detection using an ultrafast plasmonic colorimetric RT-PCR strategy.基于超快等离子体比色 RT-PCR 策略的登革热病毒检测下一代诊断检测。
Anal Chim Acta. 2023 Sep 15;1274:341565. doi: 10.1016/j.aca.2023.341565. Epub 2023 Jun 26.
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Epidemiology of dengue reported in the World Health Organization's Western Pacific Region, 2013-2019.
2013-2019 年世界卫生组织西太平洋区域登革热报告流行病学。
Western Pac Surveill Response J. 2023 Mar 22;14(1):1-16. doi: 10.5365/wpsar.2023.14.1.973. eCollection 2023 Jan-Mar.
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Three highly variable genome regions of the four dengue virus serotypes can accurately recapitulate the CDS phylogeny.四种登革病毒血清型的三个高度可变基因组区域能够准确概括编码序列系统发育。
MethodsX. 2022 Sep 17;9:101859. doi: 10.1016/j.mex.2022.101859. eCollection 2022.
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MEGA11: Molecular Evolutionary Genetics Analysis Version 11.MEGA11：分子进化遗传学分析版本 11。
Mol Biol Evol. 2021 Jun 25;38(7):3022-3027. doi: 10.1093/molbev/msab120.
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Epidemiology and challenges of dengue surveillance in the WHO South-East Asia Region.世界卫生组织东南亚区域登革热监测的流行病学和挑战。
Trans R Soc Trop Med Hyg. 2021 Jun 2;115(6):583-599. doi: 10.1093/trstmh/traa158.
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Molecular phylogeny and distribution of dengue virus serotypes circulating in Nepal in 2017.2017 年尼泊尔流行的登革热病毒血清型的分子系统发育和分布。
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8
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J Vet Diagn Invest. 2020 Nov;32(6):767-775. doi: 10.1177/1040638720905833. Epub 2020 Feb 18.
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Phylogenetic analysis revealed the co-circulation of four dengue virus serotypes in Southern Thailand.系统进化分析显示，在泰国南部四种血清型登革热病毒共同流行。
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