Sens Actuators B Chem . Rapid Point-of-Care Testing for SARS-CoV-2 Virus Nucleic Acid Detection by an Isothermal and Nonenzymatic Signal Amplification System Coupled with a Lateral Flow Immunoassay Strip

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Sens Actuators B Chem


. 2021 Apr 3;129899.
doi: 10.1016/j.snb.2021.129899. Online ahead of print.
Rapid Point-of-Care Testing for SARS-CoV-2 Virus Nucleic Acid Detection by an Isothermal and Nonenzymatic Signal Amplification System Coupled with a Lateral Flow Immunoassay Strip


Mingyuan Zou ¹ , Feiya Su ¹ , Rui Zhang ¹ , Xinglu Jiang ¹ , Han Xiao ¹ , XueJiao Yan ² , Chuankun Yang ³ , Xiaobo Fan ¹ , Guoqiu Wu ^{1 3 4}



Affiliations

• PMID: 33840901
• PMCID: PMC8019248
• DOI: 10.1016/j.snb.2021.129899

Abstract

An outbreak of a new coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), began in December 2019. Accurate, rapid, convenient, and relatively inexpensive diagnostic methods for SARS-CoV-2 infection are important for public health and optimal clinical care. The current gold standard for diagnosing SARS-CoV-2 infection is reverse transcription-polymerase chain reaction (RT-PCR). However, RTPCR assays are designed for use in well-equipped laboratories with sophisticated laboratory infrastructure and highly trained technicians, and are unsuitable for use in under-equipped laboratories and in the field. In this study, we report the development of an accurate, rapid, and easy-to-implement isothermal and nonenzymatic signal amplification system (a catalytic hairpin assembly (CHA) reaction) coupled with a lateral flow immunoassay (LFIA) strip-based detection method that can detect SARSCoV-2 in oropharyngeal swab samples. Our method avoids RNA isolation, PCR amplification, and elaborate result analysis, which typically takes 6-8 hours. The entire CHA-LFIA detection method, from nasopharyngeal sampling to obtaining test results, takes less than 90 minutes. Such methods are simple and require no expensive equipment, only a simple thermostatically controlled water bath and a fluorescence reader device. We validated our method using synthetic oligonucleotides and clinical samples from 15 patients with SARS-CoV-2 infection and 15 healthy individuals. Our detection method provides a fast, simple, and sensitive (with a limit of detection (LoD) of 2,000 copies/mL) alternative to the SARS-CoV-2 RT-PCR assay, with 100% positive and negative predictive agreements.

Keywords: Catalytic hairpin assembly reaction; Lateral flow immunoassay strip; Nucleic Acid Test; rapid diagnostic test; sars-cov-2.