Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, Yamadaoka, Suita, Osaka 565-0871, Japan.
Int J Mol Med. 2011 Oct;28(4):625-8. doi: 10.3892/ijmm.2011.733. Epub 2011 Jun 27.
Dengue virus (DENV) is a mosquito-borne virus and can be transmitted to humans by mosquito vectors. Although surveillance of dengue virus-infected mosquitoes is the most effective way of controlling DENV infections, detection of DENVs in mosquitoes is limited by the low sensitivity of available detection methods. We here report a method for capturing DENV type 3 (DENV-3) from mosquito cells using magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic anhydrate). The beads were incubated with cell culture medium of DENV-3-infected mosquito cells, then separated from the supernatant by applying a magnetic field and washed. Adsorption of DENV-3 on the beads was confirmed by reverse transcription-polymerase chain reaction, which detected the presence of DENV-3 genomic RNA on the beads, and Western blotting, which determined the major DENV-3 envelope protein on the beads. Therefore, this capture method may enable an improvement in DENV-3 detection.
登革热病毒(DENV)是一种通过蚊子传播的病毒,可以通过蚊子媒介传播给人类。虽然监测感染登革热病毒的蚊子是控制 DENV 感染的最有效方法,但现有的检测方法灵敏度有限,限制了对 DENVs 的检测。我们在此报告了一种使用阴离子聚合物聚(甲基乙烯基醚-马来酸酐)涂覆的磁珠从蚊子细胞中捕获登革热病毒 3 型(DENV-3)的方法。将珠子与感染 DENV-3 的蚊子细胞的细胞培养液一起孵育,然后通过施加磁场将其与上清液分离,并进行洗涤。通过逆转录-聚合酶链反应证实了 DENV-3 在珠子上的吸附,该反应检测了珠子上存在 DENV-3 基因组 RNA,通过 Western blot 确定了珠子上的主要 DENV-3 包膜蛋白。因此,这种捕获方法可能会提高 DENV-3 的检测能力。