Enzyme-linked immunosorbent assay of autoantibodies reacting with thyroid plasma membrane antigens in sera of patients with autoimmune thyroid diseases

1986 ◽  
Vol 113 (2) ◽  
pp. 255-260 ◽  
Author(s):  
Andrzej Gardas ◽  
Kathleen L. Rives

Abstract. A sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the detection of autoantibodies reacting with thyroid plasma membrane antigens has been established. Autoantibodies reacting with thyroid plasma membrane antigens were detected by the ELISA in 95% of untreated hyperthyroid Graves', 68% of antithyroid drug-treated Graves' up to four months of the therapy, in 62% of Hashimoto's thyroiditis and in 8.9% of toxic nodular goitre. The ELISA was negative in 100% healthy blood donors, 100% non-toxic nodular goitre, in 12 patients with rheumatoid arthritis, 18 patients with scleroderma and 94% of patients with systemic lupus erythematosus. The mean value of autoantibodies titre was higher in untreated hyperthyroid Graves' (1:84 000) and lowest in positive patients with autoimmune disease of non-thyroid origin (1:4000). The cross-reactivity of antimicrosomal antigen antibodies was below 10%; there was no influence of antithyroglobulin antibodies on the ELISA; and most of the autoantibodies react with plasma membrane antigens different from the TSH binding sites.

1984 ◽  
Vol 105 (4) ◽  
pp. 500-504 ◽  
Author(s):  
Andrzej Gardas ◽  
Barbara Czarnocka ◽  
Janusz Nauman

Abstract. The presence of antithyroid plasma membrane antibodies (ATMA) has been detected in 97% of patients with untreated hyperthyroid Graves' disease, 85% of methimazole treated hyperthyroid Graves' disease, 25% of Hashimoto's thyroiditis and 6.9% of patients with toxic nodular goitre. The ATMA index was negative in all healthy blood donors, in patients with non-toxic nodular goitre, with the thyrocardiac syndrome and with simple obesity. Studies of patients with non-thyroid autoimmune diseases revealed that ATMA is positive in 11% of patients with scleroderma, 17.6% of systemic lupus erythematosus and 16% of rheumatoid arthritis. The amount of immunoglobulin bound to thyroid plasma membranes after pre-incubation with serum from patients with Graves' disease varied from 4.2 to 25.2 pmoles per mg of membrane protein; these values are several times higher than the maximal binding capacity for thyrotrophin which is 1.28 pmole/mg protein. In the majority of the cases studied TSH did not significantly inhibit IgG bound from thyroid plasma membranes. Significant amounts of IgG were displaced by an excess of TSH only in three cases with untreated hyperthyroid Graves' disease.


2006 ◽  
Vol 44 (9) ◽  
pp. 3432-3434 ◽  
Author(s):  
M. Giacchino ◽  
N. Chiapello ◽  
S. Bezzio ◽  
F. Fagioli ◽  
P. Saracco ◽  
...  

1990 ◽  
Vol 68 (7) ◽  
pp. 1578-1588 ◽  
Author(s):  
Brian T. Luck ◽  
Colette Breuil ◽  
David L. Brown

An enzyme-linked immunosorbent assay (ELISA) was used to detect a sap-staining fungus, Ophiostoma piceae, and a biological-control agent, Gliocladium roseum, grown in liquid culture and in wood. A polyclonal serum prepared against whole cell fragments from broken mycelia of O. piceae detected O. piceae in liquid culture at 0.25 μg dry weight/mL; however, there was moderate cross-reactivity with G. roseum. Antiserum adsorbed on G. roseum had almost no reactivity with G. roseum but still reacted strongly with O. piceae. The specificity of these sera was verified, and the antigenic sites were localized, by immunogold labeling and electron microscopy. These studies confirmed that the adsorbed serum could differentiate between G. roseum and O. piceae and showed that the cell wall was the most reactive cellular component. These results are discussed in relation to the development of immunological probes for the detection of sap-staining and biological control fungi. Key words: polyclonal serum, enzyme-linked immunosorbent assay, immunogold labeling, sap-staining and biological control fungi, electron microscopy.


2021 ◽  
Vol 12 ◽  
Author(s):  
Bochao Liu ◽  
Ze Wu ◽  
Chaolan Liang ◽  
Jinhui Lu ◽  
Jinfeng Li ◽  
...  

Since December 2019, a novel coronavirus (SARS-CoV-2) has resulted in a global pandemic of coronavirus disease (COVID-19). Although viral nucleic acid test (NAT) has been applied predominantly to detect SARS-CoV-2 RNA for confirmation diagnosis of COVID-19, an urgent need for alternative, rapid, and sensitive immunoassays is required for primary screening of virus. In this study, we developed a smartphone-based nanozyme-linked immunosorbent assay (SP-NLISA) for detecting the specific nucleocapsid phosphoprotein (NP) of SARS-CoV-2 in 37 serum samples from 20 COVID-19 patients who were diagnosed by NAT previously. By using SP-NLISA, 28/37 (75.7%) serum samples were detected for NP antigens and no cross-reactivity with blood donors’ control samples collected from different areas of China. In a control assay using the conventional enzyme-linked immunosorbent assay (ELISA), only 7/37 (18.91%) serum samples were detected for NP antigens and no cross-reactivity with control samples. SP-NLISA could be used for rapid detection of SARS-CoV-2 NP antigen in primary screening of SARS-CoV-2 infected individuals.


1983 ◽  
Vol 29 (5) ◽  
pp. 823-827 ◽  
Author(s):  
M Ishaq ◽  
R Ali

Abstract In this enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against extractable nuclear antigens (ENA) in sera of patients with systemic lupus erythematosus (SLE), nylon is used as solid phase for antigen binding instead of the commonly used polystyrene surface. Optimal conditions for activation of the nylon beads, antigen coating, and other relevant factors have been investigated. We compared the incidence of anti-ENA antibodies in SLE, using chromogenic and fluorogenic enzyme substrates. Of SLE patients, 54% were positive for anti-ENA antibodies when chromogenic substrate was used as compared with 68% for fluorogenic substrate. Antibody activity against Sm and RNP antigens was distinguished on the basis of ribonuclease sensitivity of the RNP antigen. The method described offers advantages such as decreased background activity, increased surface area, facility for prolonged storage of antigen-coated solid phase, and miniaturization of the assay.


1996 ◽  
Vol 79 (2) ◽  
pp. 426-430 ◽  
Author(s):  
Touichi Tanaka ◽  
Hideharu Ikebuchi ◽  
Jun-Ichi Sawada ◽  
Mariko Okada ◽  
Yasumasa Kido

Abstract An easy, sensitive, competitive indirect enzyme- linked immunosorbent assay (CI-ELISA) for specti nomycin in chicken plasma was developed. Preparation of a spectinomycin-bovine serum albumin conjugate in which the hapten is linked to the carrier protein through the C-4 position is described. Antibodies raised against antigens in rabbits had excellent specificity for spectinomycin, exhibiting a cross-reactivity of 44.0% with dihydrospectinomy-cin and 13.8% with tetrahydrospectinomycin. No cross-reactivity was observed with other antibiotics. The detection limit of the CI-ELISA was 2 ng/mL (equivalent into 40 ng/mL undiluted chicken plasma) spectinomycin. Known amounts (0.1-100 μg/mL) of spectinomycin were added to chicken plasma and then analyzed. Average recoveries were 97-110%. This procedure may be used without prior extraction of samples.


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