scholarly journals Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion

2021 ◽  
Author(s):  
Marcelo S. Conzentino ◽  
Karl Forchhammer ◽  
Emanuel M Souza ◽  
Fábio O. Pedrosa ◽  
Meri B. Nogueira ◽  
...  
Keyword(s):  
Mass Production ◽  
Nucleocapsid Protein ◽  
Indirect Elisa ◽  
Humoral Response ◽  
High Yield ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Method ◽  
Assay Sensitivity ◽  
Elisa Kit ◽  
Chromatography Step

AbstractSerological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 PCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen coated plates were stable for up to 3 months at 4°C). The ELISA method described is ready to mass production and will be an additional toll to track COVID-19 cases.

scholarly journals Comparative Performance of In-House and Commercially Available ELISA Kits for the Detection of Antibody Against Newcastle Disease Virus Vaccine

2013 ◽  
Vol 22 (1-2) ◽  
pp. 55-64
Author(s):  
MMI Chowdhury ◽  
MT Islam ◽  
A Aktar ◽  
MKJ Bhuiyan ◽  
MM Kamal ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Newcastle Disease ◽  
Indirect Elisa ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Dilution ◽  
Positive Correlation ◽  
Elisa Kit

An In-house Indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibody titre against Newcastle disease virus (NDV) and was compared its sensitivity and specificity with the commercially available NDV antibody detection ELISA kit (Biocheck®, USA). The reference NDV was purified by centrifugation, ultracentrifugation and by sucrose density gradient ultracentrifugation. This purified NDV was used for coating of 96-well flat bottomed microtitre plate and to raise hyperimmune sera (known) in Fayoumi chickens. In the standardization test, the antigen dilution of 10-6 and the serum dilution of 10-3 were considered to be optimum for the present ELISA system. The correlation regression analysis was performed to construct a standard curve equation where a good positive correlation was observed (r = 0.912, n = 8, P<0.01). The equation was used to convert corrected absorbance readings of the single working dilution (1 : 1000) directly into predicted ELISA antibody activity titres. In the sensitivity and specificity test, the serum dilution of 10-5 appeared to be the highest dilution which had the maximum lowest capacity to bind with the coated antigen of the present ELISA kit and only anti-NDV serum was found to bind with the coated antigen instead of serum of IBDV in the plate which revealed the high specificity of the developed In-house Indirect ELISA kit. In a comparative study with the 80 chicken sera samples, a significant positive correlation (r = 0.901, n = 80, P<0.01) was found between In-house Indirect ELISA and commercial ELISA kit (Biocheck®, USA).DOI: http://dx.doi.org/10.3329/pa.v22i1-2.16467 Progress. Agric. 22(1 & 2): 55 - 64, 2011

scholarly journals Evaluation of an Indirect ELISA for the Diagnosis of Bovine Brucellosis

1995 ◽  
Vol 7 (4) ◽  
pp. 473-475 ◽  
Author(s):  
Francisco A. Uzal ◽  
Alejandra E. Carrasco ◽  
Susana Echaide ◽  
Klaus Nielsen ◽  
Carlos A. Robles
Keyword(s):  
Indirect Elisa ◽  
Screening Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Brucellosis ◽  
Energy Agency ◽  
Elisa Kit ◽  
Large Numbers ◽  
The Rose ◽  
Epidemiological Information ◽  
Control And Eradication

Control and eradication of bovine brucellosis is usually based on the serological detection of antibodies. In Argentina, the rose bengal test (RB) and the buffered plate antigen test (BPA) are the 2 screening tests officially recognized, whereas the 2-mercaptoethanol test (2ME) and the tube agglutination test (SAT) are the confirmatory assays currently in use. In order to improve the serological diagnosis of bovine brucellosis in Patagonia, Argentina, an indirect enzyme-linked immunosorbent assay (I-ELISA) kit distributed by the International Atomic Energy Agency was evaluated. Sera from negative nonvaccinated, negative but vaccinated, and positive animals were tested by all the above techniques. The specificity of the I-ELISA (98.9% and 97.2%) was similar to that of the BPA, RB, 2ME and complement fixation (CF) tests when used to test sera from negative nonvaccinated and negative but vaccinated animals, respectively. The sensitivity of the I-ELISA (98.7%) was higher than the BPA test (96.1%) and the CF test (95.2%). The I-ELISA kit evaluated in this study was thought to be a valuable tool for the diagnosis of bovine brucellosis in the Patagonia region where little epidemiological information is available about this disease, and where large numbers of sera should be tested to obtain such information.

Can Osteopontin Be Considered a Biomarker for Endometriosis?

2017 ◽  
Vol 68 (9) ◽  
pp. 2132-2134
Author(s):  
Daniela Roxana Albu (Matasariu) ◽  
Elena Mihalceanu ◽  
Alina Pangal ◽  
Carmen Vulpoi ◽  
Mircea Onofriescu ◽  
...  
Keyword(s):  
Serum Level ◽  
Immunosorbent Assay ◽  
Pelvic Pain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Multifactorial Disease ◽  
Elisa Kit ◽  
Healthy Women ◽  
Progesterone Treatment ◽  
Large Dispersion

Endometriosis is a multifactorial disease that is manifested by infertility and pelvic pain. The purpose of the study was to evaluate the effect of progesterone treatment on the serum level of osteopontin, a multipotent cytokine, in patients with endometriosis. The study was prospective and we evaluated osteopontin levels that were measured in the serum of 40 patients with endometriosis and 12 healthy women using a standardized Enzyme-Linked Immunosorbent Assay (ELISA) kit. Osteopontin seric levels were lower in endometriosis patients and increased after progesterone treatment. Because of the large dispersion of data even in the control group, we find the association between osteopontin and endometriosis questionable.

scholarly journals Rapid seroconversion and persistent functional IgG antibodies in severe COVID-19 patients correlates with an IL-12p70 and IL-33 signature

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ariel Munitz ◽  
L. Edry-Botzer ◽  
M. Itan ◽  
R. Tur-Kaspa ◽  
D. Dicker ◽  
...  
Keyword(s):  
Nucleocapsid Protein ◽  
Neutralizing Antibodies ◽  
Host Response ◽  
Humoral Response ◽  
Rapid Onset ◽  
Response Analysis ◽  
Receptor Binding Domain ◽  
Igg Antibodies ◽  
Viral Receptor ◽  
Iga Antibodies

AbstractDespite ongoing efforts to characterize the host response toward SARS-CoV-2, a major gap in our knowledge still exists regarding the magnitude and duration of the humoral response. Analysis of the antibody response in mild versus moderate/severe patients, using our new developed quantitative electrochemiluminescent assay for detecting IgM/IgA/IgG antibodies toward SARS-CoV-2 antigens, revealed a rapid onset of IgG/IgA antibodies, specifically in moderate/severe patients. IgM antibodies against the viral receptor binding domain, but not against nucleocapsid protein, were detected at early stages of the disease. Furthermore, we observed a marked reduction in IgM/IgA antibodies over-time. Adapting our assay for ACE2 binding-competition, demonstrated that the presence of potentially neutralizing antibodies is corelated with IgG/IgA. Finally, analysis of the cytokine profile in COVID-19 patients revealed unique correlation of an IL-12p70/IL33 and IgG seroconversion, which correlated with disease severity. In summary, our comprehensive analysis has major implications on the understanding and monitoring of SARS-CoV-2 infections.

scholarly journals Recombinant nucleocapsid protein-based IgG enzyme-linked immunosorbent assay for the serological diagnosis of SARS

2005 ◽  
Vol 125 (2) ◽  
pp. 181-186 ◽  
Author(s):  
Masayuki Saijo ◽  
Toshio Ogino ◽  
Fumihiro Taguchi ◽  
Shuetsu Fukushi ◽  
Tetsuya Mizutani ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Nucleocapsid Protein ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recombinant Nucleocapsid Protein

scholarly journals Indirect Enzyme-Linked Immunosorbent Assay based on the nucleocapsid protein of SARS-like coronaviruses

2009 ◽  
Vol 24 (2) ◽  
pp. 146-151 ◽  
Author(s):  
Jun-fa Yuan ◽  
Yan Li ◽  
Hua-jun Zhang ◽  
Peng Zhou ◽  
Zhen-hua Ke ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Nucleocapsid Protein ◽  
Enzyme Linked Immunosorbent Assay

Mass production of multi-wall carbon nanotubes by metal dusting process with high yield

2011 ◽  
Vol 46 (5) ◽  
pp. 716-721 ◽  
Author(s):  
H. Ghorbani ◽  
A.M. Rashidi ◽  
S. Rastegari ◽  
S. Mirdamadi ◽  
M. Alaei
Keyword(s):  
Carbon Nanotubes ◽  
Mass Production ◽  
High Yield ◽  
Metal Dusting ◽  
Multi Wall Carbon Nanotubes

scholarly journals Evaluation of an enzyme immunoassay for clinical diagnosis of neurocysticercosis in symptomatic patients

2010 ◽  
Vol 43 (6) ◽  
pp. 647-650 ◽  
Author(s):  
Reynaldo Mendes de Carvalho Junior ◽  
Dorcas Lamounier Costa ◽  
Savyo Carvalho Soares ◽  
Carlos Henrique Nery Costa
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Sensitivity And Specificity ◽  
Cerebral Spinal Fluid ◽  
Taenia Solium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Disease ◽  
Multivariate Logistic Regression ◽  
Human Central Nervous System ◽  
Elisa Kit

INTRODUCTION: Neurocysticercosis is an infection of the human central nervous system caused by the metacestode larvae of Taenia solium. Neurocysticercosis is the most common parasitic disease in developing countries. Epilepsy is the most common clinical manifestation. Difficulties in confirming the diagnosis motivated the evaluation of the enzyme-linked immunosorbent assay on cerebral spinal fluid (CSF). METHODS: Twenty-two patients with NCC and 44 control patients were studied. CSF was analyzed using a commercial ELISA kit developed for NCC. Sensitivity and specificity were measured and a multivariate logistic regression was performed. RESULTS: Sensitivity and specificity of ELISA were 31.8% and 100%, respectively, with accuracy of 77.3%. Only the size of the lesions proved to be important for performance of the test. CONCLUSIONS: The results showed that ELISA contributes to the diagnosis of neurocysticercosis if the result is negative or if the patient has a lesion of 2 cm or more.

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