Aonuma Hiroka, Sombié Aboubacar, Li Jian-Chiuan, Ote Manabu, Saiki Erisha, Ichimura Hidetoshi, Iizuka Itoe, Odagawa Taichi, Sakurai Tatsuya, Yamaji Kayoko, Saijo Masayuki, Chen Chun-Hong, Badolo Athanase, Kanuka Hirotaka
Department of Tropical Medicine, The Jikei University School of Medicine, Tokyo, Japan.
Center for Medical Entomology, The Jikei University School of Medicine, Tokyo, Japan.
PLoS One. 2025 Sep 12;20(9):e0332245. doi: 10.1371/journal.pone.0332245. eCollection 2025.
Dengue is one of the neglected tropical diseases (NTDs) transmitted by Aedes mosquitoes and continues to spread globally. When mosquitoes are infected with dengue virus (DENV), virus-derived DNA (vDNA) is generated in mosquitoes, which subsequently contributes to their immune response. We traced the generation and presence of dengue virus type 2 (DENV-2) vDNA in experimentally infected cultured mosquito cells and Aedes mosquitoes, and notably, in wild mosquitoes collected in Burkina Faso. Detection of vDNA was achieved using a method incorporating loop-mediated isothermal amplification (LAMP), specifically, a LAMP-based vDNA detection method (vDNA-LAMP). The LAMP reaction, using primers targeting a segment of the NS5 region of DENV-2, detected vDNA from crude DNA extracted from experimentally infected cultured cells and Aedes mosquitoes. Detection revealed that the amount of DENV-2 vDNA generated in infected cells was relatively low; nevertheless, vDNA-LAMP enabled successful detection. The timing and quantity of vDNA generation in cultured cells were associated with the initial number of viral particles introduced during infection. Furthermore, vDNA-LAMP was applied to detect dengue virus vDNA in wild mosquitoes in dengue-endemic regions. This resulted in the successful detection of DENV-2 vDNA in field-collected mosquitoes, indicating that a proportion of wild mosquitoes in Burkina Faso harbored DENV-2 vDNA. Mapping these vDNA-positive mosquitoes allowed the identification of areas where infected mosquitoes and/or their progeny were likely present. These findings provide insights into the dynamics of DENV-2 vDNA in natural environments and underscore the potential of vDNA-LAMP as a tool for tracing vDNA in wild mosquitoes, which are responsible for transmitting viral infections.
登革热是由伊蚊传播的被忽视的热带病之一，且仍在全球范围内蔓延。当蚊子感染登革病毒（DENV）时，病毒衍生的DNA（vDNA）会在蚊子体内产生，随后有助于其免疫反应。我们追踪了2型登革病毒（DENV-2）vDNA在实验感染的培养蚊子细胞和伊蚊中的产生及存在情况，尤其在布基纳法索采集的野生蚊子中。使用一种结合环介导等温扩增（LAMP）的方法，即基于LAMP的vDNA检测方法（vDNA-LAMP）实现了vDNA的检测。LAMP反应使用靶向DENV-2 NS5区域片段的引物，检测了从实验感染的培养细胞和伊蚊中提取的粗DNA中的vDNA。检测发现，感染细胞中产生的DENV-2 vDNA量相对较低；然而，vDNA-LAMP仍能成功检测。培养细胞中vDNA产生的时间和数量与感染期间引入的病毒颗粒初始数量有关。此外，vDNA-LAMP被应用于检测登革热流行地区野生蚊子中的登革病毒vDNA。这导致在野外采集的蚊子中成功检测到DENV-2 vDNA，表明布基纳法索的一部分野生蚊子携带DENV-2 vDNA。绘制这些vDNA阳性蚊子的分布图，能够识别可能存在感染蚊子及其后代的区域。这些发现为DENV-2 vDNA在自然环境中的动态变化提供了见解，并强调了vDNA-LAMP作为追踪野生蚊子中vDNA的工具的潜力，而野生蚊子是病毒感染传播的媒介。
