A rapid and sensitive one-pot platform integrating fluorogenic RNA aptamers and CRISPR-Cas13a for visual detection of monkeypox virus.

The recent global upsurge in Monkeypox virus (MPXV) outbreaks underscores the critical need for rapid and precise diagnostic solutions, particularly in resource-constrained settings. The gold standard diagnostic method, qRT-PCR, is hindered by its time-consuming nature, requirement for nucleic acid purification, expensive equipment, and the need for highly trained personnel. Traditional CRISPR/Cas fluorescence assays, relying on trans-cleavage of ssDNA/RNA reporters labeled with costly fluorophores and quenchers, pose challenges that limit their widespread application, especially for point-of-care testing (POCT). In this study, we utilized a cost-effective and stable fluorogenic RNA aptamer (Mango III), specifically binding and illuminating the fluorophore TO3-3 PEG-Biotin Fluorophore (TO3), as a reporter for Cas13a trans-cleavage activity. We propose a comprehensive strategy integrating RNA aptamer, recombinase-aided amplification (RAA), and CRISPR-Cas13a systems for the molecular detection of MPXV target. Leveraging the inherent collateral cleavage properties of the Cas13a system, we established high-sensitivity and specificity assays to distinguish MPXV from other Orthopoxviruses (OPVs). A streamlined one-pot protocol was developed to mitigate aerosol contamination risks. Our aptamer-coupled RAA-Cas13a one-pot detection method achieved a Limit of Detection (LoD) of 4 copies of target MPXV DNA in just 40 min. Validation using clinical MPX specimens confirmed the rapid and reliable application of our RAA-Cas13a-Apt assays without nucleic acid purification procedure, highlighting its potential as a point-of-care testing solution. These results underscore the user-friendliness and effectiveness of our one-pot RAA-Cas13a-Apt diagnostic platform, poised to revolutionize disease detection and management.

Whether the digital divide widens the income gap between China's regions?

Based on the panel data of 280 prefecture-level cities in China from 2014 to 2018, we construct the digital divide index from three aspects including "access divide", "use divide" and "efficiency divide", using Thiel index to measure income inequality and study the impact of the digital divide on income inequality between different regions. We apply the two-stage spatial DID combined IV model. In the first stage, we introduce the "Internet + government affairs" policy to construct the spatial DID model, so as to obtain the estimated value of the digital divide index. When the estimated value is substituted into the two-stage regression as an instrumental variable, it is found that every 1unit increase in the digital divide will widen the income divide by 0.134 units. This conclusion is still robust under the replacement of Theil index and change of weights. The results of heterogeneity analysis show that digital divide has a more significant impact on the income divide in eastern China in terms of regions. It is found after distinguishing different types of digital divide that the other two types of digital divides have the most significant impact than income divide, 0.034 units higher than that of the traditional. Finally, we introduce the mechanism variables from three levels of "access divide", "use divide" and "efficiency divide", thus verifying the effect of information asymmetry, effect of human capital differentiation and effect of delaying industrial upgrade. Then we put forward the following policy recommendations: in order to reduce the deterioration effect of digital divide on income distribution, it is necessary to improve infrastructure, enhance the degree of digitalization of human capital and optimize the industrial structure.