Gold Nanoparticles-Amplified Indirect Competitive Enzyme-Linked Immunosorbent Assay of BDE-121 Using a Monoclonal Antibody

2021 ◽  
Vol 21 (10) ◽  
pp. 5036-5043
Author(s):  
Yan Qiao ◽  
Qing-Yun Cai
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Gold Nanoparticles ◽  
Immunosorbent Assay ◽  
Inhibitory Concentration ◽  
High Sensitivity ◽  
High Specificity ◽  
Gas Chromatography Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
Good Recovery

In this study, we developed a monoclonal antibody against 2,3’,4,5’,6-pentabromodiphenylether (BDE-121) using a synthesized hapten, and established an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA), using gold nanoparticles, to amplify the signal. The monoclonal antibody showed high specificity, with a half inhibitory concentration (IC50) value of 2.78 ng/mL, towards BDE-121. The developed IC-ELISA exhibited high sensitivity and stability as well as good recovery. The intra-assay deviation is below 6.8% and the inter-assay deviations range from 6.5% to 8.7%. The assay of the actual samples was found to be consistent with those of gas chromatography/mass spectrometry (GC/MS).

scholarly journals Interpretations of Antibody Responses toSalmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (4) ◽  
pp. 550-555 ◽  
Author(s):  
Patrick L. McDonough ◽  
Richard H. Jacobson ◽  
John F. Timoney ◽  
Ahmed Mutalib ◽  
David C. Kradel ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Sensitivity ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Department Of Agriculture ◽  
Trace Back ◽  
Elisa Format ◽  
Commercial Poultry ◽  
Computer Controlled

ABSTRACT Many regulatory and diagnostic programs for the detection ofSalmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a humanS. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemicS. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. entericaEnteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

Comparative Assessment of Fumonisin in Grain-Based Foods by ELISA, GC-MS, and HPLC

1994 ◽  
Vol 57 (2) ◽  
pp. 169-172 ◽  
Author(s):  
JAMES J. PESTKA ◽  
JUAN I. AZCONA-OLIVERA ◽  
RONALD D. PLATTNER ◽  
FIORENZA MINERVINI ◽  
M. BRUNO DOKO ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Food Samples ◽  
Comparative Assessment ◽  
The Other ◽  
Gas Chromatography Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cereal Samples ◽  
Related Compounds ◽  
Positive Results

Seventy-one (71) food samples were analyzed for the mycotoxin fumonisin by a monoclonal antibody based competitive enzyme-linked immunosorbent assay (ELISA). Fumonisins were detected primarily in corn-based products with 7/12, 2/2 and 1/3 and 1/7 yellow cornmeal, blue cornmeal, corn muffin mix, and mixed grain cereal samples yielding positive results, respectively. When the positive samples and randomly selected negative samples were assessed by other methods, correlations (r values) between ELISA and gas chromatography-mass spectrometry (GC-MS), ELISA and high-pressure liquid chromatography (HPLC) and GC-MS and HPLC were 0.478 (p < 0.05), 0.512 (p < 0.05), and 0.946 (p < 0.01), respectively. The results suggested that although the immunoassay could be used for screening of fumonisin in food samples, higher estimates were attained by ELISA than by the other two methods particularly in the more contaminated samples. These observations may result from differences in sample preparation among the methods or because of the presence of structurally related compounds in extracts that are detectable by ELISA but not the other two methods.

scholarly journals Synergistic Use of Gold Nanoparticles (AuNPs) and “Capillary Enzyme-Linked Immunosorbent Assay (ELISA)” for High Sensitivity and Fast Assays

Sensors ◽  
2017 ◽  
Vol 18 (2) ◽  
pp. 55 ◽  
Author(s):  
Wan-Joong Kim ◽  
Hyo Cho ◽  
Bongjin Jeong ◽  
Sangwon Byun ◽  
JaeDoo Huh ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Immunosorbent Assay ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay

scholarly journals Steroid metabolites for diagnosing and predicting clinicopathological features in cortisol-producing adrenocortical carcinoma.

2020 ◽  
Author(s):  
Sawako Suzuki ◽  
Tomoki Minamidate ◽  
Akina Shiga ◽  
Yutarou Ruike ◽  
Kazuki Ishiwata ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Steroid Hormone ◽  
High Sensitivity ◽  
High Specificity ◽  
Clinicopathological Features ◽  
Gas Chromatography Mass Spectrometry ◽  
Large Area ◽  
Urine Metabolites ◽  
Positive Correlations ◽  
Adrenocortical Carcinomas

Abstract Background: Approximately 60% of adrenocortical carcinomas (ACC) are functional, and Cushing’s syndrome is the most frequent diagnosis that has been revealed to have a particularly poor prognosis. Since 30% of ACC present steroid hormone-producing disorganization, measurement of steroid metabolites in suspected ACC is recommended. Previous reports demonstrated that steroid hormone precursors or their urine metabolites, which can be assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS) or gas chromatography mass spectrometry (GC-MS) respectively, are useful for distinguishing ACC from cortisol-producing adenomas (CPA); however, despite high precision, LC-MS/MS and GC-MS require a highly trained team, are expensive and have limited capacity. Methods: Here, we examined 12 serum steroid metabolites using an immunoassay, which is a more rapid and less costly method than LC-MS/MS, in cortisol-producing ACC and CPA. Further, the correlation of each steroid metabolite to the classification stage and pathological status in ACC was analyzed. Results: Reflecting disorganized steroidogenesis, the immunoassay revealed that all basal levels of steroid precursors were significantly increased in cortisol-producing ACC compared to CPA; in particular, 17-hydroxypregnenolone (glucocorticoid and androgen precursor) and 11-deoxycorticosterone (mineralocorticoid precursor) showed a large area under the ROC curve with high sensitivity and specificity when setting the cut-off at 1.78 ng/ml and 0.4 mg/ml, respectively. Additionally, a combination of androstenedione and DHEAS also showed high specificity with high accuracy. In cortisol-producing ACC, 11-deoxycortisol (glucocorticoid precursor) showed significant positive correlations with predictive prognostic factors used in ENSAT classification, while testosterone showed significant positive correlations to the Ki67-index in both men and women. Conclusion: Less expensive and more widely available RIA and ECLIA may also biochemically distinguish ACC from CPA and may predict the clinicopathological features of ACC.

scholarly journals Steroid metabolites for diagnosing and predicting clinicopathological features in cortisol-producing adrenocortical carcinoma.

2020 ◽  
Author(s):  
Sawako Suzuki ◽  
Tomoki Minamidate ◽  
Akina Shiga ◽  
Yutarou Ruike ◽  
Kazuki Ishiwata ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Steroid Hormone ◽  
High Sensitivity ◽  
High Specificity ◽  
Clinicopathological Features ◽  
Gas Chromatography Mass Spectrometry ◽  
Large Area ◽  
Urine Metabolites ◽  
Positive Correlations ◽  
Adrenocortical Carcinomas

Abstract Background: Approximately 60% of adrenocortical carcinomas (ACC) are functional, and Cushing’s syndrome is the most frequent diagnosis that has been revealed to have a particularly poor prognosis. Since 30% of ACC present steroid hormone-producing disorganization, measurement of steroid metabolites in suspected ACC is recommended. Previous reports demonstrated that steroid hormone precursors or their urine metabolites, which can be assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS) or gas chromatography mass spectrometry (GC-MS) respectively, are useful for distinguishing ACC from cortisol-producing adenomas (CPA); however, despite high precision, LC-MS/MS and GC-MS require a highly trained team, are expensive and have limited capacity. Methods: Here, we examined 12 serum steroid metabolites using an immunoassay, which is a more rapid and less costly method than LC-MS/MS, in cortisol-producing ACC and CPA. Further, the correlation of each steroid metabolite to the classification stage and pathological status in ACC was analyzed. Results: Reflecting disorganized steroidogenesis, the immunoassay revealed that all basal levels of steroid precursors were significantly increased in cortisol-producing ACC compared to CPA; in particular, 17-hydroxypregnenolone (glucocorticoid and androgen precursor ) and 11-deoxycorticosterone (mineralocorticoid precursor) showed a large area under the ROC curve with high sensitivity and specificity when setting the cut-off at 1.78 ng/ml and 0.4 mg/ml, respectively. Additionally, a combination of androstenedione and DHEAS also showed high specificity with high accuracy. In cortisol-producing ACC, 11-deoxycortisol (glucocorticoid precursor) showed significant positive correlations with predictive prognostic factors used in ENSAT classification, while testosterone showed significant positive correlations to the Ki67-index in both men and women.Conclusion: Less expensive and more widely available RIA and ECLIA may also biochemically distinguish ACC from CPA and may predict the clinicopathological features of ACC.

scholarly journals Steroid metabolites for diagnosing and predicting clinicopathological features in cortisol-producing adrenocortical carcinoma.

2020 ◽  
Author(s):  
Sawako Suzuki ◽  
Tomoki Minamidate ◽  
Akina Shiga ◽  
Yutarou Ruike ◽  
Kazuki Ishiwata ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Steroid Hormone ◽  
High Sensitivity ◽  
High Specificity ◽  
Clinicopathological Features ◽  
Gas Chromatography Mass Spectrometry ◽  
Large Area ◽  
Urine Metabolites ◽  
Positive Correlations ◽  
Adrenocortical Carcinomas

Abstract Background: Approximately 60% of adrenocortical carcinomas (ACC) are functional, and Cushing’s syndrome is the most frequent diagnosis that has been revealed to have a particularly poor prognosis. Since 30% of ACC present steroid hormone-producing disorganization, measurement of steroid metabolites in suspected ACC is recommended. Previous reports demonstrated that steroid hormone precursors or their urine metabolites, which can be assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS) or gas chromatography mass spectrometry (GC-MS) respectively, are useful for distinguishing ACC from cortisol-producing adenomas (CPA); however, despite high precision, LC-MS/MS and GC-MS require a highly trained team, are expensive and have limited capacity.Methods: Here, we examined 12 serum steroid metabolites using an immunoassay, which is a more rapid and less costly method than LC-MS/MS, in cortisol-producing ACC and CPA. Further, the correlation of each steroid metabolite to the classification stage and pathological status in ACC was analyzed.Results: Reflecting disorganized steroidogenesis, the immunoassay revealed that all basal levels of steroid precursors were significantly increased in cortisol-producing ACC compared to CPA; in particular, 17-hydroxypregnenolone (glucocorticoid and androgen precursor) and 11-deoxycorticosterone (mineralocorticoid precursor) showed a large area under the ROC curve with high sensitivity and specificity when setting the cut-off at 1.78 ng/ml and 0.4 mg/ml, respectively. Additionally, a combination of androstenedione and DHEAS also showed high specificity with high accuracy. In cortisol-producing ACC, 11-deoxycortisol (glucocorticoid precursor) showed significant positive correlations with predictive prognostic factors used in ENSAT classification, while testosterone showed significant positive correlations to the Ki67-index in both men and women.Conclusion: Less expensive and more widely available RIA and ECLIA may also biochemically distinguish ACC from CPA and may predict the clinicopathological features of ACC.

scholarly journals Steroid metabolites for diagnosing and predicting clinicopathological features in cortisol-producing adrenocortical carcinoma

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Sawako Suzuki ◽  
Tomoki Minamidate ◽  
Akina Shiga ◽  
Yutarou Ruike ◽  
Kazuki Ishiwata ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Steroid Hormone ◽  
High Sensitivity ◽  
High Specificity ◽  
Clinicopathological Features ◽  
Gas Chromatography Mass Spectrometry ◽  
Large Area ◽  
Urine Metabolites ◽  
Positive Correlations ◽  
Adrenocortical Carcinomas

Abstract Background Approximately 60% of adrenocortical carcinomas (ACC) are functional, and Cushing’s syndrome is the most frequent diagnosis that has been revealed to have a particularly poor prognosis. Since 30% of ACC present steroid hormone-producing disorganization, measurement of steroid metabolites in suspected ACC is recommended. Previous reports demonstrated that steroid hormone precursors or their urine metabolites, which can be assessed using liquid chromatography tandem mass spectrometry (LC-MS/MS) or gas chromatography mass spectrometry (GC-MS) respectively, are useful for distinguishing ACC from cortisol-producing adenomas (CPA); however, despite high precision, LC-MS/MS and GC-MS require a highly trained team, are expensive and have limited capacity. Methods Here, we examined 12 serum steroid metabolites using an immunoassay, which is a more rapid and less costly method than LC-MS/MS, in cortisol-producing ACC and CPA. Further, the correlation of each steroid metabolite to the classification stage and pathological status in ACC was analyzed. Results Reflecting disorganized steroidogenesis, the immunoassay revealed that all basal levels of steroid precursors were significantly increased in cortisol-producing ACC compared to CPA; in particular, 17-hydroxypregnenolone (glucocorticoid and androgen precursor) and 11-deoxycorticosterone (mineralocorticoid precursor) showed a large area under the ROC curve with high sensitivity and specificity when setting the cut-off at 1.78 ng/ml and 0.4 mg/ml, respectively. Additionally, a combination of androstenedione and DHEAS also showed high specificity with high accuracy. In cortisol-producing ACC, 11-deoxycortisol (glucocorticoid precursor) showed significant positive correlations with predictive prognostic factors used in ENSAT classification, while testosterone showed significant positive correlations to the Ki67-index in both men and women. Conclusion Less expensive and more widely available RIA and ECLIA may also biochemically distinguish ACC from CPA and may predict the clinicopathological features of ACC.

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