tetano
Editor, Senior Moderator
Appl Microbiol Biotechnol
. 2022 Aug 17.
doi: 10.1007/s00253-022-12113-8. Online ahead of print.
Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening
Ashish Kumar Srivastava[SUP] 1 [/SUP], Avinash Gupta[SUP] 1 [/SUP], Deepika Chauhan[SUP] 1 [/SUP], Ramesh Chand Meena[SUP] 1 [/SUP], Ragumani Sugadev[SUP] 1 [/SUP], Malleswara Rao Eslavath[SUP] 1 [/SUP], Harshita Gupta[SUP] 1 [/SUP], Karuna[SUP] 1 [/SUP], Sayar Singh[SUP] 1 [/SUP], Yamini Singh[SUP] 1 [/SUP], R P Tiwari[SUP] 2 [/SUP], Veena Kohli[SUP] 2 [/SUP], Rajeev Varshney[SUP] 1 [/SUP], Lilly Ganju[SUP] 3 [/SUP]
Affiliations
Abstract
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test "CoroSuchak," with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit "CoroSuchak" is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening.
Keywords: Antibody detection; COVID-19; ELISA; Human; IgG; Mass screening.
. 2022 Aug 17.
doi: 10.1007/s00253-022-12113-8. Online ahead of print.
Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening
Ashish Kumar Srivastava[SUP] 1 [/SUP], Avinash Gupta[SUP] 1 [/SUP], Deepika Chauhan[SUP] 1 [/SUP], Ramesh Chand Meena[SUP] 1 [/SUP], Ragumani Sugadev[SUP] 1 [/SUP], Malleswara Rao Eslavath[SUP] 1 [/SUP], Harshita Gupta[SUP] 1 [/SUP], Karuna[SUP] 1 [/SUP], Sayar Singh[SUP] 1 [/SUP], Yamini Singh[SUP] 1 [/SUP], R P Tiwari[SUP] 2 [/SUP], Veena Kohli[SUP] 2 [/SUP], Rajeev Varshney[SUP] 1 [/SUP], Lilly Ganju[SUP] 3 [/SUP]
Affiliations
- PMID: 35976427
- DOI: 10.1007/s00253-022-12113-8
Abstract
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test "CoroSuchak," with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit "CoroSuchak" is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening.
Keywords: Antibody detection; COVID-19; ELISA; Human; IgG; Mass screening.