Slip formation of a high-density droplet array for nucleic acid quantification by digital LAMP with a random-access system.

Abstract

Digital nucleic acid analysis (digital NAA) is an important tool for the precise quantification of nucleic acids. Various microfluidic-based approaches for digital NAA have been developed, but most methods require complex auxiliary control instruments, cumbersome device fabrication, or inconvenient preparation processes. A SlipChip is a microfluidic device that can generate and manipulate liquid partitions through simple movements of two microfluidic plates in close contact. However, the traditional SlipChip requires accurate alignment of microfeatures on different plates; therefore, the dimensions of the microwells and density of partitions can be constrained. Here, we developed a droplet array SlipChip (da-SlipChip) that can form droplets of various sizes at high density in a single slipping step. This process does not require precise overlapping microfeatures on different plates; therefore, the design flexibility and partition density can be significantly increased. We quantified SARS-CoV-2 nucleic acids extracted from the COVID-19 pseudovirus by digital loop-mediated isothermal amplification (LAMP) on a da-SlipChip with 21 696 of 0.25 nL droplets, and the results were in good agreement with those of the commercial digital PCR method of Stilla. Furthermore, we demonstrated a random-access system with a single-throughput fluorescence imager and a stackable thermal control instrument with nine independent heating modules. This random-access system with the da-SlipChip can greatly improve the total throughput and flexibility for digital isothermal nucleic acid quantification and significantly reduce the total waiting time.

• Authors: Weiyuan Lyu, Jiajie Zhang, Yan Yu, Lei Xu, Feng Shen
• Journal: Lab on a chip (Lab Chip) Vol. 21 Issue 16 Pg. 3086-3093 (08 21 2021) ISSN: 1473-0189 [Electronic] England
• PMID: 34160518 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Nucleic Acids

Topics

• COVID-19
• Humans
• Molecular Diagnostic Techniques
• Nucleic Acid Amplification Techniques
• Nucleic Acids
• SARS-CoV-2