A fully automated microfluidic PCR-array system for rapid detection of multiple respiratory tract infection pathogens

Abstract

Rapid and accurate identification of respiratory tract infection pathogens is of utmost importance for clinical diagnosis and treatment, as well as prevention of pathogen transmission. To meet this demand, a microfluidic chip-based PCR-array system, Onestart, was developed. The Onestart system uses a microfluidic chip packaged with all the reagents required, and the waste liquid is also collected and stored on the chip. This ready-to-use system can complete the detection of 21 pathogens in a fully integrated manner, with sample lysis, nucleic acid extraction/purification, and real-time PCR sequentially implemented on the same chip. The entire analysis process is completed within 1.5 h, and the system automatically generates a test report. The lower limit-of-detection (LOD) of the Onestart assay was determined to be 1.0 × 10 3 copies•mL −1 . The inter-batch variation of cycle threshold (Ct) values ranged from 0.08% to 0.69%, and the intra-batch variation ranged from 0.9% to 2.66%. Analytical results of the reference sample mix showed a 100% specificity of the Onestart assay. The analysis of batched clinical samples showed consistency of the Onestart assay with real-time PCR. With its ability to provide rapid, sensitive, and specific detection of respiratory tract infection pathogens, application of the Onestart system will facilitate timely clinical management of respiratory tract infections and effective prevention of pathogen transmission.

High—throughput and automated screening for COVID-19

“…Onestart was able to complete the sample-in-answer-out process including lysis of samples, nucleic acid extraction and amplification and result output in a fully automated manner. The study reported consistent results with real-time PCR with 100% specificity in 21 different pathogens (27).…”

“…After sample preparation, this platform performed a fully automated process from sample-in to answer-out using centrifugal force for nucleic acid separation in a microfluidic disc ( 26 ). For the detection of multiple respiratory tract pathogens including SARS-CoV-2, a microfluidic chip-based PCR-array system, Onestart, was developed using magnetic force for nucleic acid purification ( 27 ). Onestart was able to complete the sample-in-answer-out process including lysis of samples, nucleic acid extraction and amplification and result output in a fully automated manner.…”

Micro/nanotechnology-inspired rapid diagnosis of respiratory infectious diseases

“…Beyond diagnosing a single infectious disease, Huang et al . [ 196 ] developed Onestart, which is a microfluidic chip-based PCR-array system to diagnose respiratory infection. The Onestart can detect 21 different respiratory pathogens in an integrated manner that processes from sample dissolution to real-time PCR at once and has the advantage of easy operation and short processing time.…”

A microfluidic system for rapid nucleic acid analysis based on real-time convective PCR at point-of-care testing

“…In contrast, for point-of-care molecular diagnostics, simple, convenient, easy-to-operate, and even ‘sample-in, answer-out’ nucleic acid analysis can be achieved based on other concepts, for example, microfluidics (Zhuang et al 2020 ; Soni and Toley 2022 ). With a properly designed fluid network integrated with different functional units, for example, micro-valves, micro-channels, micro-reactors, and micro-actuators, POC nucleic acid analysis can be performed on a single microfluidic chip normally accompanied by a portable instrument (Pumford et al 2020 ; Huang et al 2021 ). Superior to traditional methods, point-of-care molecular diagnostics is capable of implementing immediate detection with high convenience in resource-poor settings, which is quite beneficial to efficient detection of infectious deceases, for example, COVID-19 in the global pandemic (GeneXpert 2022 ; Abbott.…”