Manually-operated, slider cassette for multiplexed molecular detection at the point of care

• Published in: Sensors and actuators. B, Chemical Chemical, 2022-10, Vol.369, p.132353, Article 132353
• Main Authors: Seok, Youngung, Yin, Qingtian, Li, Ruijie, Mauk, Michael G., Bai, Huiwen, Bau, Haim H.
• Format: Article
• Language: English
• Subjects: Amplification; Blistering; Blisters; Capillarity; Cassettes; Chambers; Disease control; Hepatitis; Hepatitis B virus (HBV); Hepatitis C; Hepatitis C virus (HCV); HIV; Housing; Human immunodeficiency virus; Human immunodeficiency virus (HIV); Loop-mediated isothermal amplification (LAMP); Microfluidics; Microprocessors; Molecular Diagnostics; Multiplexed; Multiplexing; Nucleic acids; Public health; Reagents; Sample to answer
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2022.132353

Effective control of epidemics, individualized medicine, and new drugs with virologic response-dependent dose and timing require, among other things, simple, inexpensive, multiplexed molecular detection platforms suitable for point of care and home use. Herein, we describe our progress towards developing such a platform that includes sample lysis, nucleic acid isolation, concentration, purification, and amplification. Our diagnostic device comprises a sliding component that houses the nucleic acid isolation membrane and a housing containing three amplification reaction chambers with dry stored reagents, blisters with buffers and wash solutions, and absorption pads to facilitate capillarity pull and waste storage. After sample introduction, the user slides the slider within the housing from one station to another to carry out various unit operations. The slider motion induces blisters to discharge their contents, effectuating washes, and eventual elution of captured nucleic acids into reaction chambers. The slider cassette mates with a processor that incubates isothermal amplification but can also be made to operate instrumentation-free. We demonstrate our cassette’s utility for the co-detection of the human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). These three blood-borne pathogens co-infect many people worldwide with severe personal and public health consequences. •Slider cassette from sample processing to real-time molecular detection in point-of-care testing has been developed.•HHBV, HCV, and HIV from human plasma are able to be co-detected in a single analysis.•Sliding component, isolation membrane, blisters, absorption pad, and three amplification chambers are combined.•Multiplexed microfluidics, heating, and slider moving are strongly supported by the simulation results.•RReliable results from 30 to 300000 IU of HBV virus and from 5 μL to 100 μL range of human plasma were obtained.