tetano
Editor, Senior Moderator
Microbiol Spectr
. 2021 Nov 24;e0067221.
doi: 10.1128/Spectrum.00672-21. Online ahead of print.
Usefulness of ELISA Using Total Antibody against Plant-Expressed Recombinant Nucleocapsid Protein of SARS-CoV-2
Misbah Tariq[SUP] #[/SUP][SUP] 1 [/SUP], Jian Hur[SUP] #[/SUP][SUP] 2 [/SUP], Jun-Won Seo[SUP] 1 [/SUP], Da Young Kim[SUP] 1 [/SUP], Na Ra Yun[SUP] 1 [/SUP], You Mi Lee[SUP] 1 [/SUP], Mi-Seon Bang[SUP] 1 [/SUP], Seong Yeon Hwang[SUP] 1 [/SUP], Choon-Mee Kim[SUP] 3 [/SUP], Ju-Hyung Lee[SUP] 4 [/SUP], Kyoung-Ho Song[SUP] 5 [/SUP], Hyunju Lee[SUP] 6 [/SUP], Jongtak Jung[SUP] 5 [/SUP], Ji Young Park[SUP] 6 [/SUP], Hong Bin Kim[SUP] 5 [/SUP], Eu Suk Kim[SUP] 5 [/SUP], Sangmin Lee[SUP] 7 [/SUP], Dong-Min Kim[SUP] 1 [/SUP]
Affiliations
Abstract
Here, we aimed to investigate the diagnostic value of a serological assay using the nucleocapsid protein developed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection and evaluated its performance using three commercial enzyme-linked immunosorbent assays (ELISAs), namely, Standard E 2019 novel coronavirus disease (COVID-19) total antibody (Ab) ELISA (SD Biosensor), and EDI novel coronavirus COVID-19 IgG and IgM ELISA. A recombinant nucleocapsid protein (rNP) was expressed from plants and Escherichia coli for the detection of serum total Ab. We prospectively collected 141 serum samples from 32 patients with reverse transcription-PCR (RT-PCR)-confirmed COVID-19 and determined the sensitivity and dynamics of their total Ab response. Specificity was evaluated using 158 prepandemic samples. To validate the assays, we evaluated the performance using two different cutoff values. The sensitivity and specificity for each assay were as follows: 92.91% and 94.30% (plant-rNP), 83.69% and 98.73% (SD Biosensor), 75.89% and 98.10% (E. coli-rNP), 76.47% and 100% (EDI-IgG), and 80.39% and 80% (EDI-IgM). The plant-based rNP showed the highest sensitivity and area under the receiver operating characteristic (ROC) curve (0.980) among all the assays (P < 0.05). The seroconversion rate for total Ab increased sequentially with disease progression, with a sensitivity of 100% after 10 to 12 days of post-symptom onset (PSO) for both rNP-plant-based and SD Biosensor ELISAs. After 2 weeks of PSO, the seroconversion rates were >80% and 100% for EDI-IgM and EDI-IgG ELISA, respectively. Seroconversion occurred earlier with rNP plant-based ELISA (5 days PSO) compared with E. coli-based (7 days PSO) and SD Biosensor (8 days PSO) ELISA. We determined that rNP produced in plants enables the robust detection of SARS-CoV-2 total Abs. The assay can be used for serosurvey and complementary diagnosis of COVID-19. IMPORTANCE At present, the principal diagnostic methods for COVID-19 comprise the identification of viral nucleic acid by genetic approaches, including PCR-based techniques or next-generation sequencing. However, there is an urgent need for validated serological assays which are crucial for the understanding of immune responses against SARS-CoV-2. In this study, a highly sensitive and specific serological antibody assay was developed for the detection of SARS-CoV-2 with an overall accuracy of 93.56% using a recombinant nucleoprotein expressed from plants.
Keywords: COVID-19; E. coli; ELISA; SARS-CoV-2; nucleoprotein; plant; total antibody.
. 2021 Nov 24;e0067221.
doi: 10.1128/Spectrum.00672-21. Online ahead of print.
Usefulness of ELISA Using Total Antibody against Plant-Expressed Recombinant Nucleocapsid Protein of SARS-CoV-2
Misbah Tariq[SUP] #[/SUP][SUP] 1 [/SUP], Jian Hur[SUP] #[/SUP][SUP] 2 [/SUP], Jun-Won Seo[SUP] 1 [/SUP], Da Young Kim[SUP] 1 [/SUP], Na Ra Yun[SUP] 1 [/SUP], You Mi Lee[SUP] 1 [/SUP], Mi-Seon Bang[SUP] 1 [/SUP], Seong Yeon Hwang[SUP] 1 [/SUP], Choon-Mee Kim[SUP] 3 [/SUP], Ju-Hyung Lee[SUP] 4 [/SUP], Kyoung-Ho Song[SUP] 5 [/SUP], Hyunju Lee[SUP] 6 [/SUP], Jongtak Jung[SUP] 5 [/SUP], Ji Young Park[SUP] 6 [/SUP], Hong Bin Kim[SUP] 5 [/SUP], Eu Suk Kim[SUP] 5 [/SUP], Sangmin Lee[SUP] 7 [/SUP], Dong-Min Kim[SUP] 1 [/SUP]
Affiliations
- PMID: 34817278
- DOI: 10.1128/Spectrum.00672-21
Abstract
Here, we aimed to investigate the diagnostic value of a serological assay using the nucleocapsid protein developed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection and evaluated its performance using three commercial enzyme-linked immunosorbent assays (ELISAs), namely, Standard E 2019 novel coronavirus disease (COVID-19) total antibody (Ab) ELISA (SD Biosensor), and EDI novel coronavirus COVID-19 IgG and IgM ELISA. A recombinant nucleocapsid protein (rNP) was expressed from plants and Escherichia coli for the detection of serum total Ab. We prospectively collected 141 serum samples from 32 patients with reverse transcription-PCR (RT-PCR)-confirmed COVID-19 and determined the sensitivity and dynamics of their total Ab response. Specificity was evaluated using 158 prepandemic samples. To validate the assays, we evaluated the performance using two different cutoff values. The sensitivity and specificity for each assay were as follows: 92.91% and 94.30% (plant-rNP), 83.69% and 98.73% (SD Biosensor), 75.89% and 98.10% (E. coli-rNP), 76.47% and 100% (EDI-IgG), and 80.39% and 80% (EDI-IgM). The plant-based rNP showed the highest sensitivity and area under the receiver operating characteristic (ROC) curve (0.980) among all the assays (P < 0.05). The seroconversion rate for total Ab increased sequentially with disease progression, with a sensitivity of 100% after 10 to 12 days of post-symptom onset (PSO) for both rNP-plant-based and SD Biosensor ELISAs. After 2 weeks of PSO, the seroconversion rates were >80% and 100% for EDI-IgM and EDI-IgG ELISA, respectively. Seroconversion occurred earlier with rNP plant-based ELISA (5 days PSO) compared with E. coli-based (7 days PSO) and SD Biosensor (8 days PSO) ELISA. We determined that rNP produced in plants enables the robust detection of SARS-CoV-2 total Abs. The assay can be used for serosurvey and complementary diagnosis of COVID-19. IMPORTANCE At present, the principal diagnostic methods for COVID-19 comprise the identification of viral nucleic acid by genetic approaches, including PCR-based techniques or next-generation sequencing. However, there is an urgent need for validated serological assays which are crucial for the understanding of immune responses against SARS-CoV-2. In this study, a highly sensitive and specific serological antibody assay was developed for the detection of SARS-CoV-2 with an overall accuracy of 93.56% using a recombinant nucleoprotein expressed from plants.
Keywords: COVID-19; E. coli; ELISA; SARS-CoV-2; nucleoprotein; plant; total antibody.