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Usefulness of ELISA Using Total Antibody against Plant-Expressed Recombinant Nucleocapsid Protein of SARS-CoV-2.
Tariq, Misbah; Hur, Jian; Seo, Jun-Won; Kim, Da Young; Yun, Na Ra; Lee, You Mi; Bang, Mi-Seon; Hwang, Seong Yeon; Kim, Choon-Mee; Lee, Ju-Hyung; Song, Kyoung-Ho; Lee, Hyunju; Jung, Jongtak; Park, Ji Young; Kim, Hong Bin; Kim, Eu Suk; Lee, Sangmin; Kim, Dong-Min.
  • Tariq M; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Hur J; Department of Infectious Disease Internal Medicine, College of Medicine, Yeungnam University, Gyeongsan, Republic of Korea.
  • Seo JW; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Kim DY; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Yun NR; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Lee YM; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Bang MS; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Hwang SY; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Kim CM; Premedical Science, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
  • Lee JH; Department of Preventive Medicine, Jeonbuk National University Medical School, Jeonju, Republic of Korea.
  • Song KH; Department of Internal Medicine, Seoul National University Bundang Hospitalgrid.412480.b, College of Medicine, Seongnam, Republic of Korea.
  • Lee H; Department of Pediatrics, Seoul National University Bundang Hospitalgrid.412480.b, Seongnam, Republic of Korea.
  • Jung J; Department of Internal Medicine, Seoul National University Bundang Hospitalgrid.412480.b, College of Medicine, Seongnam, Republic of Korea.
  • Park JY; Department of Pediatrics, Seoul National University Bundang Hospitalgrid.412480.b, Seongnam, Republic of Korea.
  • Kim HB; Department of Internal Medicine, Seoul National University Bundang Hospitalgrid.412480.b, College of Medicine, Seongnam, Republic of Korea.
  • Kim ES; Department of Internal Medicine, Seoul National University Bundang Hospitalgrid.412480.b, College of Medicine, Seongnam, Republic of Korea.
  • Lee S; BioApplications Inc., Pohang, Republic of Korea.
  • Kim DM; Department of Internal Medicine, College of Medicine, Chosun Universitygrid.254187.dgrid.464555.3grid.254187.dgrid.464555.3, Gwangju, Republic of Korea.
Microbiol Spectr ; 9(3): e0067221, 2021 12 22.
Article in English | MEDLINE | ID: covidwho-1532977
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Plant Proteins / Enzyme-Linked Immunosorbent Assay / Nucleocapsid Proteins / COVID-19 Testing / COVID-19 / Antibodies, Viral Type of study: Diagnostic study / Experimental Studies / Prognostic study Limits: Humans Language: English Journal: Microbiol Spectr Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Plant Proteins / Enzyme-Linked Immunosorbent Assay / Nucleocapsid Proteins / COVID-19 Testing / COVID-19 / Antibodies, Viral Type of study: Diagnostic study / Experimental Studies / Prognostic study Limits: Humans Language: English Journal: Microbiol Spectr Year: 2021 Document Type: Article