Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 6, Pages 2926-2933Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c04858
Keywords
-
Categories
Funding
- National Natural Science Foundation of China [61927819, 81827808, 20170012, 62105177]
- National Key Research and Development Program of China [2018YFA0704004]
- Tsinghua University Spring Breeze Fund [20201080510]
- Beijing Lab Foundation [20201550018]
- Tsinghua Autonomous Research Foundation [20194180031, 20201080058]
- China Postdoctoral Science Foundation [2020M680591]
- Shuimu Tsinghua Scholar Program [2019SM150]
Ask authors/readers for more resources
This study presents a rapid, highly sensitive, and label-free pathogen assay system based on a solid-phase self-interference RPA chip (SiSA-chip). The system offers high efficiency, sensitivity, specificity, and low cost for pathogen DNA identification, making it promising for fast identification and diagnosis of infectious diseases, including SNP genotyping.
Recombinase polymerase amplification (RPA) is a useful pathogen identification method. Several label-free detection methods for RPA amplicons have been developed in recent years. However, these methods still lack sensitivity, specificity, efficiency, or simplicity. In this study, we propose a rapid, highly sensitive, and label-free pathogen assay system based on a solid-phase self-interference RPA chip (SiSA-chip) and hyperspectral interferometry. The SiSA-chips amplify and capture RPA amplicons on the chips, rather than irrelevant amplicons such as primer dimers, and the SiSA-chips are then analysed by hyperspectral interferometry. Optical length increases of SiSA-chips are used to demonstrate RPA detection results, with a limit of detection of 1.90 nm. This assay system can detect as few as six copies of the target 18S rRNA gene of Plasmodium falciparum within 20 min, with a good linear relationship between the detection results and the concentration of target genes (R-2 = 0.9903). Single nucleotide polymorphism (SNP) genotyping of the dhfr gene of Plasmodium falciparum is also possible using the SiSA-chip, with as little as 1% of mutant gene distinguished from wild-type loci (m/wt). This system offers a high-efficiency (20 min), high-sensitivity (6 copies/reaction), high-specificity (1% m/wt), and low-cost (similar to 1/50 of fluorescence assays for RPA) diagnosis method for pathogen DNA identification. Therefore, this system is promising for fast identification of pathogens to help diagnose infectious diseases, including SNP genotyping.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available