School of Biomedical Engineering, International University, Ho Chi Minh City 700000, Vietnam.
Vietnam National University, Ho Chi Minh City 700000, Vietnam.
Anal Methods. 2023 Aug 17;15(32):3991-3999. doi: 10.1039/d3ay00756a.
The dengue virus, once transmitted to people through a mosquito bite, causes an infectious disease called dengue fever. Dengue fever can develop into two fatal syndromes, namely dengue shock syndrome and dengue hemorrhagic fever. The existing strategies for detecting dengue infection mainly employ serological immunoassays and a real time PCR technique. Along with the positive features of efficiency, accuracy, and reproducibility, these procedures are limited by being time-consuming, costly, requiring special equipment for analysis, and unable to be carried out at the point-of-care level. Herein, we developed a colorimetric nanosensor for detecting dengue virus in clinical samples that is rapid, accurate, sensitive, and less expensive. The sensing platform relies on the specific binding between the DNA-conjugated AuNPs and genomic RNA of dengue, which results in the DNA-RNA heteroduplex structure formation that turns the gold colloid's ruby red color to blue due to the nano-aggregation in an acidic environment, which can be detected by the naked eye or measuring the absorbance. The DNA probe was designed to bind to a genomic RNA conserved region recognized in all four dengue serotypes. Dengue virus serotype 1 was utilized as a framework for virus detection; the designed nanosensor exhibited great specificity and selectivity, with the detection limit of ∼1 pg μL (∼1.66 × 10 RNA copies per reaction) and time of analysis of about 1 h including the RNA extraction step. The proposed colorimetric nanosensor offers an alternative tool for specific and highly sensitive detection of dengue that eliminates the requirement for thermal cycling and primer sets in PCR-based assays.
登革病毒,经蚊子叮咬传播给人类后,引发一种名为登革热的传染病。登革热可能发展成两种致命综合征,即登革休克综合征和登革出血热。目前用于检测登革感染的策略主要采用血清学免疫测定法和实时 PCR 技术。虽然这些方法具有效率高、准确性好、重现性强等优点,但也存在耗时、昂贵、需要专门的分析设备,以及无法在护理点进行的局限性。在此,我们开发了一种用于检测临床样本中登革病毒的比色纳米传感器,该传感器具有快速、准确、灵敏和廉价的特点。该传感平台依赖于 DNA 修饰的金纳米颗粒与登革病毒基因组 RNA 的特异性结合,导致 DNA-RNA 异源双链体结构的形成,在酸性环境中由于纳米聚集,使金胶体的红宝石红色变为蓝色,可通过肉眼或测量吸光度进行检测。DNA 探针设计用于结合所有四种登革血清型中公认的基因组 RNA 保守区域。登革病毒 1 型被用作病毒检测的框架;所设计的纳米传感器具有出色的特异性和选择性,检测限约为 1 pg μL(每个反应约 1.66 × 10 个 RNA 拷贝),包括 RNA 提取步骤在内的分析时间约为 1 小时。这种比色纳米传感器为特异性和高灵敏度检测登革病毒提供了一种替代工具,消除了基于 PCR 测定法中热循环和引物对的要求。