Abstract
BackgroundThe novel coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which has quickly spread worldwide since its outbreak in December 2019. One of the primary measures for controlling the spread of SARS-CoV-2 infection is an accurate assay for its diagnosis. SARS-CoV-2 real-time PCR kits suffer from some limitations, including false-negative results in the clinic. Therefore, there is an urgent need for the development of a rapid antibody test kit for COVID-19 diagnosis.MethodsThe nuclear capsid protein (N) and spike protein 1 (S1) fragments of SARS-CoV-2 were expressed in Escherichia coli, and rapid antibody-based tests for the diagnosis of SARS-CoV-2 infection were developed. To evaluate their clinical applications, the serum from COVID-19 patients, suspected COVID-19 patients, recovering COVID-19 patients, patients with general fever or pulmonary infection, doctors and nurses who worked at the fever clinic, and health professionals was analyzed by the rapid antibody test kits. The serum from patients infected with Mycoplasma pneumoniae and patients with respiratory tract infection was further analyzed to test its cross-reactivity with other respiratory pathogens.ResultsA 47 kDa N protein and 67 kDa S1 fragment of SARS-CoV-2 were successfully expressed, purified, and renatured. The rapid antibody test with recombinant N protein showed higher positive rate than the rapid IgM antibody test with recombinant S1 protein. Clinical evaluation showed that the rapid antibody test kit with recombinant N protein had 88.56 % analytical sensitivity and 97.42 % specificity for COVID-19 patients, 53.48 % positive rate for suspected COVID-19 patients, 57.14 % positive rate for recovering COVID-19 patients, and 0.5−0.8 % cross-reactivity with other respiratory pathogens. The analytical sensitivity of the kit did not significantly differ in COVID-19 patients with different disease courses (p < 0.01).ConclusionsThe rapid antibody test kit with recombinant N protein has high specificity and analytical sensitivity, and can be used for the diagnosis of SARS-CoV-2 infection combined with RT-PCR.
Highlights
The novel coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which has quickly spread worldwide since its outbreak in December 2019
Patients were treated at the Pulmonary Hospital of Lanzhou (Lanzhou, China) and Nanjing Gulou Hospital (Nanjing, China); 88 patients had general fever or pulmonary infection (COVID-19-negative) in the outpatient service of 940 Hospital of Chinese People’s Liberation Army Joint Logistic Support, 124 had Mycoplasma pneumoniae infection, 185 patients with respiratory tract infection were treated at the 940 Hospital, 48 doctors and nurses worked in the fever outpatient service of the 940 hospital (COVID-19-negative), and 56 health professionals underwent a medical examination in the health examination center of Gansu Province Tumor Hospital (Gansu Province, China)
Analysis was performed using SPSS 23.0, and the Cloning of the N gene and spike protein 1 (S1) fragment of SARS‐CoV‐2 The N gene (1257 bp) and S1 fragment (1800 bp) of SARS-CoV-2 were successfully cloned into pET28a and pET30a vectors, respectively
Summary
The novel coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which has quickly spread worldwide since its outbreak in December 2019. SARS-CoV-2 real-time PCR kits suffer from some limitations, including false-negative results in the clinic. There is an urgent need for the development of a rapid antibody test kit for COVID-19 diagnosis. The novel coronavirus disease 2019 (COVID-19) has quickly spread around the world since its outbreak in December 2019. After the outbreak of COVID-19, COVID-19 RT-PCR kits were developed and used for the clinical diagnosis of SARS-CoV-2 infection [2]. Screening asymptomatic carriers is very important for controlling the spread of COVID-19 To this end, there is an urgent need to develop a rapid, simple, specific, sensitive, and accurate test for COVID-19 diagnosis
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