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

SENSORS AND ACTUATORS B-CHEMICAL

卷:342

Article

Zou, Mingyuan¹  Su, Feiya¹  Zhang, Rui¹  Jiang, Xinglu¹  Xiao, Han¹  Yan, XueJiao⁴  Yang, Chuankun²  Fan, Xiaobo¹  Wu, Guoqiu^1,2,3 

[1] Southeast Univ, Med Sch, Nanjing 210009, Peoples R China

[2] Southeast Univ, Zhongda Hosp, Ctr Clin Lab Med, Nanjing 210009, Peoples R China

[3] Southeast Univ, Jiangsu Prov Key Lab Crit Care Med, Nanjing, Peoples R China

[4] Nanjing Med Univ, Affiliated Changzhou Peoples Hosp 2, Changzhou 213000, Jiangsu, Peoples R China

关键词: sars-cov-2;    Nucleic acid test;    Rapid diagnostic test;    Catalytic hairpin assembly reaction;    Lateral flow immunoassay strip;   

DOI  :  10.1016/j.snb.2021.129899

来源: Elsevier

【 摘 要 】

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 h. The entire CHA-LFIA detection method, from nasopharyngeal sampling to obtaining test results, takes less than 90 min. 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 2000 copies/mL) alternative to the SARSCoV-2 RT-PCR assay, with 100 % positive and negative predictive agreements.

【 授权许可】

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