scholarly journals Development of an Enzyme-Linked Immunosorbent Assay-Based Test with a Cocktail of Nucleocapsid and Spike Proteins for Detection of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific Antibody

2008 ◽  
Vol 16 (2) ◽  
pp. 241-245 ◽  
Author(s):  
Luis G. Giménez ◽  
Jose Rojas ◽  
Almudena Rojas ◽  
Joaquín Mendoza ◽  
Ana G. Camacho
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Immunosorbent Assay ◽  
Immunofluorescence Assay ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recombinant Polypeptide ◽  
Igm Antibody ◽  
Set Up ◽  
The Individual ◽  
Recombinant Polypeptides

ABSTRACT A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.

scholarly journals Recombinant Truncated Nucleocapsid Protein as Antigen in a Novel Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Severe Acute Respiratory Syndrome Coronavirus Infection

2007 ◽  
Vol 14 (2) ◽  
pp. 146-149 ◽  
Author(s):  
Fuxun Yu ◽  
Mai Quynh Le ◽  
Shingo Inoue ◽  
Futoshi Hasebe ◽  
Maria del Carmen Parquet ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Immunosorbent Assay ◽  
Nucleocapsid Protein ◽  
Seroconversion Rate ◽  
Disease Onset ◽  
Immunoglobulin M ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igm Antibody ◽  
Specificity And Sensitivity

ABSTRACT We report the development of an immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for severe acute respiratory syndrome coronavirus (SARS-CoV) by using recombinant truncated SARS-CoV nucleocapsid protein as the antigen. The newly developed MAC-ELISA had a specificity and sensitivity of 100% as evaluated by using sera from healthy volunteers and patients with laboratory-confirmed SARS. Using serial serum samples collected from SARS patients, the times to seroconversion were determined by IgM antibody detection after SARS-CoV infection. The median time to seroconversion detection was 8 days (range, 5 to 17 days) after disease onset, and the seroconversion rates after the onset of illness were 33% by the first week, 97% by the second week, and 100% by the third week. Compared with the results of our previous report on the detection of IgG, the median seroconversion time by IgM detection was 3 days earlier and the seroconversion rate by the second week after the illness for IgM was significantly higher than by IgG assay. Our results indicating that the IgM response appears earlier than IgG after SARS-CoV infection in consistent with those for other pathogens. Our newly developed MAC-ELISA system offers a new alternative for the confirmation of SARS-CoV infection.

scholarly journals Diagnostic Performance of Three Serological Assays for Anti-SARS-CoV-2 Antibody Detection

2021 ◽  
Vol 5 (4) ◽  
pp. 162-165
Author(s):  
Shabnam Dildar ◽  
◽  
Asma Danish ◽  
Mehjabeen Imam ◽  
Arshi Naz ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Diagnostic Performance ◽  
Immunofluorescence Assay ◽  
Lateral Flow ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Perfect Agreement ◽  
N Protein ◽  
Assay Performance ◽  
Antibody Elisa

Abstract: Objective: To evaluate the diagnostic performance of Electrochemiluminescence (ECLIA) enzyme linked immunosorbent (ELISA) and lateral flow Immunofluorescence (LFIA) for anti-SARS-COV-2 antibody detection. Materials and Methods: Sensitivity was calculated with convalescent plasma (CP) donor’s samples. Specificity was checked by using pre-pandemic October 2019 samples. All samples were tested for anti-SARS-COV-2 antibody by using Electrochemiluminescence (ECLIA), Enzyme Linked Immunosorbent Assay (ELISA) and Lateral flow Immunofluorescence (LFIA) assay. Results: Total 55 patients were included, 45 patients were CP donors and 10 were Pre-Pandemic October 2019 samples archived from our blood bank. The ECLIA-total antibody, ELISA-IgG and LLFIA-IgG were positive in 41 (91.1%), 34 (75.5%) and 44 (97.75%) respectively. The highest sensitivity was observed for LFIA with highest specificity among all three assays. There was almost perfect agreement between LFIA and ECLIA (k=0.936, p<0.001) but there was fair agreement between LFIA and ELISA (k=0.412, p=0.001) and ECLIA and ELISA (k=0.357, p=0.001). Conclusion: The LFIA showed a higher sensitivity and specificity in comparison with ECLIA and ELISA. It might be due to fact that LFIA detect antibody against ncleocapsid and spike protein as well of SARS- COV-2 virus, while ECLIA and ELISA detects antibodies only against “N” Protein of SARS- COV-2 virus. Keywords: Convalescent plasma donors, Lateral flow Immunofluorescence assay, Electrochemiluminescence assay, Enzyme linked immunosorbent assay, Performance.

scholarly journals Use of the COOH Portion of the Nucleocapsid Protein in an Antigen-Capturing Enzyme-Linked Immunosorbent Assay for Specific and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus

2005 ◽  
Vol 12 (3) ◽  
pp. 474-476 ◽  
Author(s):  
Maofeng Qiu ◽  
Jin Wang ◽  
Hongxia Wang ◽  
Zeliang Chen ◽  
Erhei Dai ◽  
...  
Keyword(s):  
Amino Acids ◽  
Severe Acute Respiratory Syndrome ◽  
Epidemiological Study ◽  
Immunosorbent Assay ◽  
Nucleocapsid Protein ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sars Coronavirus ◽  
Specific Test ◽  
N Protein

ABSTRACT Antibody detection with a recombinant COOH portion of the severe acute respiratory syndrome (SARS) coronavirus nucleocapsid (N) protein, N13 (amino acids 221 to 422), was demonstrated to be more specific and sensitive than that with the full-length N protein, and an N13-based antigen-capturing enzyme-linked immunosorbent assay providing a convenient and specific test for serodiagnosis and epidemiological study of SARS was developed.

Simultaneous Analysis of T-2 Toxin and HT-2 Toxin by an Indirect Enzyme-Linked Immunosorbent Assay

1987 ◽  
Vol 70 (4) ◽  
pp. 657-661
Author(s):  
Titan S L Fan ◽  
Yi-Chun Xu ◽  
Fun Sun Chu
Keyword(s):  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Simultaneous Analysis ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Toxin A ◽  
Toxin Concentration ◽  
Mixed Sample ◽  
Analytical Recovery ◽  
The Individual

Abstract An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of HT-2 toxin in the presence or absence of T-2 toxin is described. In the indirect ELISA, the relative cross-reactivities of antibodies against T-2 toxin (anti-T-2) with T-2 toxin and HT-2 toxin were 1 and 0.1, whereas anti-HT-2 cross-reactivities with T-2 toxin and HT-2 toxin were 0.33 and 1, respectively. Using such relationships, a formula was established that could be used to calculate the individual toxin concentration in a mixed sample after experimentally analyzing for T-2 and HT-2 toxins in the 2 indirect ELISAs. This method was tested by analyzing urine samples spiked with HT-2 toxin alone and samples spiked with both T-2 toxin and HT-2 toxin. A cleanup protocol for treatment of urine samples before ELISA was also established. The overall analytical recovery of HT-2 toxin when it was added at concentrations of 0.1-10 parts per billion (ppb) to the urine samples was ca 89%. When both T-2 and HT-2 toxins were added to the urine samples at equal concentrations of 0.5 to 5.0 ppb, their recoveries were 112 and 109%, respectively.

scholarly journals Comparison of Commercial Enzyme-Linked Immunosorbent Assay and Immunofluorescence Assay for Diagnosis of Acute Rickettsia typhi Infections

2020 ◽  
Vol 20 (2) ◽  
pp. 93-99 ◽  
Author(s):  
Dewi Lokida ◽  
Pratiwi Sudarmono ◽  
Herman Kosasih ◽  
Deni Pepy Butar-butar ◽  
Gustiani Salim ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rickettsia Typhi ◽  
Commercial Enzyme
Sign in / Sign up

Export Citation Format

Share Document