登革热病毒
多路复用
核糖核酸
分子信标
清脆的
生物
生物传感器
计算生物学
DNA
分子生物学
病毒学
登革热
寡核苷酸
遗传学
基因
生物化学
作者
Min Zhong,Jinbo Liu,Jie Wu,Jin-Qian Li,Nini Luo,Chuanlong Zhu,Rui Liu,Qianfeng Xia,Huangxian Ju
标识
DOI:10.1016/j.snb.2022.132011
摘要
Dengue caused by dengue virus (DENV) is a highly pathogenic viral disease. Early detection of pathogens is very important for patient treatment and pathogen control. This work designed a dual signal amplification strategy for sensitive fluorescent detection of DENV RNA. The dual signal amplification was achieved by using a target-triggered hybridization chain reaction (HCR) to connect multiplex proximity ligation activated CRISPR-Cas12a (pCRISPR-Cas12a). The HCR was triggered by the recognition of DENV RNA to a hairpin probe (H3), which led to a product to initiate the continuous hybridization of a couple of hairpins. Here one of the hairpins (H2) was designed to contain two domains ( a and b ), and domains a and b from two H2 formed a proximity ligation sequence (PLS) to connect the multiplex pCRISPR-Cas12a onto the HCR product, in which domain a activated the Cas12a, and domain b enhanced the enzymatic activity by about 5 times. Upon the enzymatic cleavage of single-stranded DNA labeled with a fluorophore and quencher at each end (ssDNA-FQ), a strong fluorescent signal was produced for DENV RNA analysis, which showed a linear range from 1 pM to 10 nM and a detection limit of 51 fM. The excellent performance of the proposed homogenous fluorescent assay with high sensitivity and superior accuracy endowed this strategy broad application prospects in clinical disease diagnosis. • A universal fluorescence biosensing strategy was developed for highly sensitive detection of Dengue virus RNA. • The parallel connection of proximity sequence enhanced CRISPR-Cas12a was achieved through target-triggered HCR. • This proposed homogenous detection method showed a wide detection range and 50 fM-level detection limit. • This proposed strategy possesses good accuracy, high precision, and good extendability for target analysis.
科研通智能强力驱动
Strongly Powered by AbleSci AI