scholarly journals Recombinant Human Tissue Transglutaminase ELISA for the Diagnosis of Gluten-sensitive Enteropathy

1999 ◽  
Vol 45 (12) ◽  
pp. 2142-2149 ◽  
Author(s):  
Miklós Sárdy ◽  
Uwe Odenthal ◽  
Sarolta Kárpáti ◽  
Mats Paulsson ◽  
Neil Smyth
Keyword(s):  
Guinea Pig ◽  
Tissue Transglutaminase ◽  
Serum Samples ◽  
Serological Tests ◽  
Specificity And Sensitivity ◽  
Gluten Sensitive Enteropathy ◽  
One Step ◽  
The Mean ◽  
Monkey Esophagus ◽  
Endomysium Antibody

Abstract Background: Tissue transglutaminase (TGc) has recently been identified as the major, if not the sole, autoantigen of gluten-sensitive enteropathy (GSE). We developed and validated an ELISA based on the human recombinant antigen and compared it to existing serological tests for GSE [guinea pig TGc ELISA and endomysium antibody (EMA) test]. Methods: Human TGc was expressed in the human embryonic kidney cell line 293-EBNA as a C-terminal fusion protein with the eight-amino acid Strep-tag II allowing one-step purification via streptavidin affinity chromatography. We carried out ELISA assays for IgA antibodies against TGc using calcium-activated human and guinea pig TGc. The sera were also tested on monkey esophagus sections by indirect immunofluorescence for IgA EMA. We examined 71 serum samples from patients with GSE (38 with celiac disease, 33 with dermatitis herpetiformis), including 16 on therapy, and 53 controls. Results: The human TGc could be expressed and purified as an active enzyme giving a single band on a Coomassie-stained gel. The mean intra- and interassay CVs for the human TGc ELISA were 3.2% and 9.2%, respectively. The area under the ROC curve was 0.999. The specificity and sensitivity were 98.1% (95% confidence interval, 95.7–100%) and 98.2% (95.9–100%), respectively. Conclusions: The human TGc ELISA was somewhat superior to the guinea pig TGc ELISA, and was as specific and sensitive as the EMA test. The human TGc-based ELISA is the method of choice for easy and noninvasive screening and diagnosis of GSE.

Comparison of a tissue transglutaminase ELISA with the endomysium antibody test in the diagnosis of gluten-sensitive enteropathy

2000 ◽  
Vol 38 (5) ◽  
pp. 357-364 ◽  
Author(s):  
M. Sárdy ◽  
S. Kárpáti ◽  
F. Péterfy ◽  
K. Rásky ◽  
E. Tomsits ◽  
...  
Keyword(s):  
Tissue Transglutaminase ◽  
Antibody Test ◽  
Gluten Sensitive Enteropathy ◽  
Endomysium Antibody

scholarly journals Rapid Serological Tests for SARS-COV-2: Diagnostic Performance of Four Commercial Assays

2020 ◽  
Author(s):  
Sérgio Monteiro de Almeida ◽  
Regiane Nogueira Spalanzani ◽  
Meri Bordignon Nogueira ◽  
Beatriz Sanada Spiri ◽  
Barbara Maria Cavalli ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Rapid Test ◽  
Nucleic Acid Amplification ◽  
Control Group ◽  
Serum Samples ◽  
Serological Tests ◽  
Test Kit ◽  
Healthy Control ◽  
One Step ◽  
Substantial Heterogeneity

Abstract Background. This study aimed to assess the diagnostic performance of lateral flow immunochromatographic assays (LFA) of four different manufacturers to identify SARS-CoV-2 antibodies (IgM, IgG or total), comparing them with the nucleic acid amplification test (NAAT) or clinical defined (definite or probable SARS-CoV-2 infection respectively). Methods. 119 serum samples were randomly selected by convenience and distributed in the groups: (1) Group with SARS-CoV-2 infection [n=82; RT-qPCR positive (definite, n=70), and probable (n=12)]; (2) other diseases [n= 27; other viruses identified (n=8), SARS of other etiologies (n=19)]; (3) healthy control group (n=10). LFA essays of four manufacturers were compared: MedTest Coronavírus (COVID-19) IgG/IgM (MedLevensohn, Brazil); COVID-19 IgG/IgM ECO Test (Ecodiagnóstica, Brazil); Camtech COVID-19 IgM/IgG Rapid Test Kit (Camtech Diagnostics Pte Ltd, Singapore); and one Step COVID-19 Test for total antibodies (Guangzhou Wondfo Biotech Co, China).Results. The four tests studied showed high diagnostic performance characteristics for the diagnoses of definite or probable SARS-CoV-2 infection. The best measures were for the Wondfo test: sensitivity (86.59%; 95%CI, 77.26-93.11%); specificity (100%; 90.51-100%); DOR (257; 60-1008); LR+ (33.43; 4.82-231.85); LR− (0.13; 0.08 - 0.23); accuracy (90.76%; 84.06- 95.29%); Matthews Correlation coefficient (MCC) 0.82. Although considering only the probable SARS-CoV-2 infection (PCR-) cases, all the kits studied showed limited values.Conclusion. Our data demonstrate the excellent performance of LFA for the diagnoses of definite or probable SARS-CoV-2 infection. There was substantial heterogeneity in sensitivities of IgM and IgG antibodies among the manufacturers. LFA tests cannot replace molecular diagnostics, but should be used as additional screening tool.

scholarly journals Rapid Serological Tests for SARS-COV-2: Diagnostic Performance of Four Commercial Assays

2020 ◽  
Author(s):  
Sérgio Monteiro de Almeida ◽  
Regiane Nogueira Spalanzani ◽  
Meri Bordignon Nogueira ◽  
Beatriz Sanada Spiri ◽  
Barbara Maria Cavalli ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Rapid Test ◽  
Nucleic Acid Amplification ◽  
Control Group ◽  
Serum Samples ◽  
Serological Tests ◽  
Test Kit ◽  
Healthy Control ◽  
One Step ◽  
Substantial Heterogeneity

Abstract Background. This study aimed to assess the diagnostic performance of lateral flow immunochromatographic assays (LFA) of four different manufacturers to identify SARS-CoV-2 antibodies (IgM, IgG or total), comparing them with the nucleic acid amplification test (NAAT) or clinical defined (definite or probable SARS-CoV-2 infection respectively). Methods. 119 serum samples were randomly selected by convenience and distributed in the groups: (1) Group with SARS-CoV-2 infection [n=82; RT-qPCR positive (definite, n=70), and probable (n=12)]; (2) other diseases [n= 27; other viruses identified (n=8), SARS of other etiologies (n=19)]; (3) healthy control group (n=10). LFA essays of four manufacturers were compared: MedTest Coronavírus (COVID-19) IgG/IgM (MedLevensohn, Brazil); COVID-19 IgG/IgM ECO Test (Ecodiagnóstica, Brazil); Camtech COVID-19 IgM/IgG Rapid Test Kit (Camtech Diagnostics Pte Ltd, Singapore); and one Step COVID-19 Test for total antibodies (Guangzhou Wondfo Biotech Co, China).Results. The four tests studied showed high diagnostic performance characteristics for the diagnoses of definite or probable SARS-CoV-2 infection. The best measures were for the Wondfo test: sensitivity (86.59%; 95%CI, 77.26-93.11%); specificity (100%; 90.51-100%); DOR (257; 60-1008); LR+ (33.43; 4.82-231.85); LR− (0.13; 0.08 - 0.23); accuracy (90.76%; 84.06- 95.29%); Matthews Correlation coefficient (MCC) 0.82. Although considering only the probable SARS-CoV-2 infection (PCR-) cases, all the kits studied showed limited values.Conclusion. Our data demonstrate the excellent performance of LFA for the diagnoses of definite or probable SARS-CoV-2 infection. There was substantial heterogeneity in sensitivities of IgM and IgG antibodies among the manufacturers. LFA tests cannot replace molecular diagnostics, but should be used as additional screening tool.

scholarly journals Celiac-Related Autoantibodies and IL-17A in Bulgarian Patients with Dermatitis Herpetiformis: A Cross-Sectional Study

Medicina ◽  
2019 ◽  
Vol 55 (5) ◽  
pp. 136
Author(s):  
Tsvetelina Velikova ◽  
Martin Shahid ◽  
Ekaterina Ivanova-Todorova ◽  
Kossara Drenovska ◽  
Kalina Tumangelova-Yuzeir ◽  
...  
Keyword(s):  
Dermatitis Herpetiformis ◽  
Tissue Transglutaminase ◽  
Serum Levels ◽  
Cross Sectional Study ◽  
Serum Samples ◽  
Cross Sectional ◽  
Gastrointestinal Complications ◽  
Gluten Sensitive Enteropathy ◽  
Diagnostic Sensitivity And Specificity ◽  
Average Serum

Background and objectives: Dermatitis herpetiformis (DH) is a blistering dermatosis, which shares common immunologic features with celiac disease (CD). The aim of the present study was to explore the performance of a panel of CD-related antibodies and IL-17A in Bulgarian patients with DH. Materials and Methods: Serum samples from 26 DH patients at mean age 53 ± 15 years and 20 healthy controls were assessed for anti-tissue transglutaminase (anti-tTG), anti-deamidated gliadin peptides (anti-DGP), anti-actin antibodies (AAA), and IL-17A by enzyme linked immuno-sorbent assay (ELISA), as well as anti-tTG, anti-gliadin (AGA), and anti-Saccharomyces cerevisiae antibodies (ASCA) using immunoblot. Results: The average serum levels of anti-tTG, anti-DGP, AGA, AAA, and the cytokine IL-17A were at significantly higher levels in patients with DH compared to the average levels in healthy persons which stayed below the cut-off value (p < 0.05). Anti-DGP and anti-tTG antibodies showed the highest diagnostic sensitivity and specificity, as well as acceptable positive and negative predictive value. None of the healthy individuals was found positive for the tested antibodies, as well as for ASCA within the DH group. All tests showed good to excellent correlations (r = 0.5 ÷ 0.9, p < 0.01). Conclusions: Although the diagnosis of DH relies on skin biopsy for histology and DIF, serologic testing of a panel of celiac-related antibodies could be employed with advantages in the diagnosing process of DH patients. Furthermore, DH patients who are positive for the investigated serologic parameters could have routine monitoring for gastrointestinal complications typical for the gluten-sensitive enteropathy.

scholarly journals Rapid Serological Tests for Sars-Cov-2: Diagnostic Performance of Four Commercial Assays

2021 ◽  
Author(s):  
Sérgio Monteiro de Almeida ◽  
Regiane Nogueira Spalanzani ◽  
Meri Bordignon Nogueira ◽  
Beatriz Sanada ◽  
Barbara Maria Cavalli ◽  
...  
Keyword(s):  
Diagnostic Performance ◽  
Rapid Test ◽  
Nucleic Acid Amplification ◽  
Control Group ◽  
Serum Samples ◽  
Serological Tests ◽  
Test Kit ◽  
Healthy Control ◽  
One Step ◽  
Substantial Heterogeneity

To assess the diagnostic performance of lateral flow immunochromatographic assays (LFA) of four different manufacturers to identify SARS-CoV-2 antibodies (IgM, IgG or total), comparing them with the nucleic acid amplification test (NAAT) or clinical defined (definite or probable SARS-CoV-2 infection respectively). Methods. 119 serum samples were randomly selected by convenience and distributed in the groups: (1) Group with SARS-CoV-2 infection [n=82; RT-qPCR positive (definite, n=70), and probable (n=12)]; (2) other diseases [n= 27; other viruses identified (n=8), SARS of other etiologies (n=19)]; (3) healthy control group (n=10). LFA essays of four manufacturers were compared: MedTest Coronavírus (COVID-19) IgG/IgM (MedLevensohn, Brazil); COVID-19 IgG/IgM ECO Test (Ecodiagnóstica, Brazil); Camtech COVID-19 IgM/IgG Rapid Test Kit (Camtech Diagnostics Pte Ltd, Singapore); and one Step COVID-19 Test for total antibodies (Guangzhou Wondfo Biotech Co, China). Results. The four tests studied showed high diagnostic performance characteristics for the diagnoses of definite or probable SARS-CoV-2 infection. The best measures were for the Wondfo test: sensitivity (86.59%; 95%CI, 77.26-93.11%); specificity (100%; 90.51-100%); DOR (257; 60-1008); LR+ (33.43; 4.82-231.85); LR− (0.13; 0.08 - 0.23); accuracy (90.76%; 84.06- 95.29%); Matthews Correlation coefficient (MCC) 0.82. Although considering only the probable SARS-CoV-2 infection (PCR-) cases, all the kits studied showed limited values. Conclusion. Our data demonstrate the excellent performance of LFA for the diagnoses of definite or probable SARS-CoV-2 infection. There was substantial heterogeneity in sensitivities of IgM and IgG antibodies among the manufacturers. LFA tests cannot replace molecular diagnostics, but should be used as additional screening tool.

scholarly journals A novel one-step quick assay for detection of SARS-COV2 antibodies across mammalian species

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11381
Author(s):  
Xianjin Zhou
Keyword(s):  
Mammalian Species ◽  
Antibody Test ◽  
Human Infection ◽  
Step Test ◽  
Serum Samples ◽  
Serological Tests ◽  
Animal Reservoir ◽  
Quick Test ◽  
One Step ◽  
The One

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) has so far infected almost a hundred of millions of people and caused more than a million of death across the world. Many serological tests have been developed to track down virus infection in community via identification of antibodies against SARS-CoV2 virus. However, the tests vary in sensitivity, specificity, complexity, and speed. Here, I developed a simple, one-step, quick test to detect antibodies against SARS-CoV2 N (scN) nucleocapsid protein via direct visualization of antigen-antibody reaction. A total of 40 serum samples of SARS-CoV2 patients were purchased from RayBiotech. A total of 50 pre-pandemic human serum samples from San Diego Blood Bank were used as negative controls. After performing the one-step quick test of these 90 serum samples, I found that 39 samples are positive for anti-scN antibodies. All of the 39 positives are from the 40 SARS-CoV2 patients, suggesting that the one-step test is more sensitive than the lateral flow immunoassay (LFIA), the most widely used rapid antibody test. None of the 50 pre-pandemic samples is positive for anti-scN antibodies, indicating that the one-step test has an excellent specificity. The one-step test takes only ~5 min to detect the antibodies; and 1 ml of Escherichia coli culture can produce reagent proteins sufficient for thousands of the tests. Since the one-step test does not need a secondary antibody, it can be used as a universal test for anti-scN antibodies across different mammalian species to track down both human infection and the animal reservoir of SARS-CoV2 virus.

scholarly journals Diagnostic Performance of Commercially Available Enzyme-Linked Immunosorbent Assay Kit in the Diagnosis of Extrapulmonary Tuberculosis

2013 ◽  
Vol 5 (01) ◽  
pp. 11-16 ◽  
Author(s):  
Manimuthu Mani Sankar ◽  
Veena Balooni ◽  
Jitendra Singh ◽  
Sarman Singh
Keyword(s):  
Developing Countries ◽  
Diagnostic Performance ◽  
Extrapulmonary Tuberculosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Pulmonary Tb ◽  
The Mean ◽  
The Individual ◽  
Individual Specificity

ABSTRACT Objectives: Antibody based serodiagnosis tests for tuberculosis (TB) was used widely in developed and developing countries. Pathozyme Myco® immunoglobulin (Ig) M, IgA, and IgG were evaluated in pulmonary TB in many studies. Materials and Methods: In this study we assessed this commercially available kit in detecting extrapulmonary TB (EPTB). Results: A total of 354 subjects were recruited for the study, of which 217 (61.2%) were EPTB patients and 137 (38.7%) were subjects with no suggestive TB. The mean age was 29.7 ± 13.7 and 31.2 ± 15.2 years, respectively for two groups. Serum samples were tested for IgM, IgA, and IgG using Pathozyme Myco® IgM, IgA, and IgG kit. The individual specificity rates of IgM, IgA, and IgG were 70.8% (95% confidence interval (CI): 62.7-77.7), 77.3% (95% CI: 68.6-83.5), and 68.6%. (95% CI: 60.4-75.7); while their sensitivity was 29% (95% CI: 23.4-35.4), 24.4% (95% CI: 19.1-30.5), and 34.5% (95% CI: 28.5-41.1); respectively. Conclusion: The serological tests either singly or in combination failed or performed poorly to diagnose EPTB.

scholarly journals Celiac Disease and Non-celiac Gluten Sensitivity Screening in Juvenile Idiopathic Arthritis Patients

2020 ◽  
Author(s):  
Günsel Kutluk ◽  
Hafize Emine Sönmez ◽  
Hülya Kaçmaz
Keyword(s):  
Juvenile Idiopathic Arthritis ◽  
Celiac Disease ◽  
Tissue Transglutaminase ◽  
Cross Sectional Study ◽  
Gluten Sensitivity ◽  
Laboratory Findings ◽  
Cross Sectional ◽  
The Mean ◽  
Autoimmune Etiology ◽  
Endomysium Antibody

INTRODUCTION: Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease in children characterized by autoimmune etiology. In previous studies, increased Celiac disease (CD) frequency was reported in patients diagnosed with JIA. In this study, CD and non-celiac gluten sensitivity (NCGS) in patients diagnosed with JIA were investigated. METHODS: Sixty-one (57.3%) JIA patients admitted to the pediatric rheumatology outpatient clinic between January 2020 and April 2020 were included in this cross-sectional study. All patients were evaluated with clinical and laboratory findings in terms of CD and NCGS. Total immunoglobulin (Ig)-A, tissue transglutaminase antibody (tTG) IgA and IgG, anti-endomysium-antibody (EMA) IgA and IgG and anti-gliadin-antibody (AGA) IgA and IgG levels were measured in all patients. RESULTS: Sixty-one JIA patients, 35 girls, were included in the study. The mean age of the patients was 11.4 ± 4.6 years, the mean age at diagnosis is 10.2 ± 3.4 years. Thirty-three patients were diagnosed with oligoarticular JIA; 18 patients with enthesitis-related arthritis, 8 patients with polyarticular JIA, and 2 patients with psoriatic arthritis. All patients were using disease-modifying antirheumatic drugs during the study. Thirty-five patients were receiving biological therapy, concomitantly. Two patients had abdominal pain, two patients had indigestion, and two patients had constipation. None of the patients had growth retardation. EMA IgA and IgG, tTG IgA and IgG, AGA IgA and IgG tests were negative in all patients. DISCUSSION AND CONCLUSION: CD and NCGS were not detected in our JIA patients. Multi-center studies may guide clinicians in under what circumstances to perform CD and NCGS screening in JIA patients.

scholarly journals Performance of six SARS-CoV-2 immunoassays in comparison with microneutralisation

2020 ◽  
Author(s):  
Anne J Jääskeläinen ◽  
Suvi Kuivanen ◽  
Eliisa Kekäläinen ◽  
Maarit J Ahava ◽  
Raisa Loginov ◽  
...  
Keyword(s):  
Respiratory Virus ◽  
University Hospital ◽  
Serum Samples ◽  
Neutralising Antibodies ◽  
Serological Tests ◽  
Specificity And Sensitivity ◽  
Novel Virus ◽  
Overall Performance ◽  
Variable Performance ◽  
Abbott Architect

There is an urgent need for reliable high-throughput serological assays for the management of the ongoing COVID-19 pandemic. Preferably, the performance of serological tests for a novel virus should be determined with clinical specimens against a gold standard, i.e. virus neutralisation. We evaluated specificity and sensitivity of six commercial immunoassays for the detection of SARS-CoV-2 IgG, IgA and IgM antibodies, including four automated assays [Abbott SARS-COV-2 IgG (CE marked), Diasorin Liaison SARS-CoV-2 S1/S2 IgG (research use only), and Euroimmun SARS-CoV-2 IgG and IgA (CE marked)], and two rapid lateral flow (immunocromatographic) tests [Acro Biotech 2019-nCoV IgG/IgM (CE marked) and Xiamen Biotime Biotechnology SARS-CoV-2 IgG/IgM (CE marked)] in comparison with a microneutralisation test (MNT). Two specimen panels from serum samples sent to Helsinki University Hospital Laboratory (HUSLAB) were compiled: the patient panel included sera from PCR confirmed COVID-19 patients, and the negative panel included sera sent for screening of autoimmune diseases and respiratory virus antibodies in 2018 and 2019. The MNT was carried out for all COVID-19 samples (70 serum samples, 62 individuals) and for 53 samples from the negative panel. Forty-one out of 62 COVID-19 patients showed neutralising antibodies with median of 11 days (range 3-51) after onset of symptoms. The specificity and sensitivity values of the commercial tests against MNT, respectively, were as follows: 95.1%/80.5% (Abbott Architect SARS-CoV-2 IgG), 94.9%/43.8% (Diasorin Liaison SARS-CoV-2 IgG), 68.3%/87.8% (Euroimmun SARS-CoV-2 IgA), 86.6%/70.7% (Euroimmun SARS-CoV-2 IgG), 74.4%/56.1% (Acro 2019-nCoV IgG), 69.5%/46.3% (Acro 2019-nCoV IgM), 97.5%/71.9% (Xiamen Biotime SARS-CoV-2 IgG), and 88.8%/81.3% (Xiamen Biotime SARS-CoV-2 IgM). This study shows variable performance values. Laboratories should carefully consider their testing process, such as a two-tier approach, in order to optimize the overall performance of SARS-CoV-2 serodiagnostics.

Immunologic Assessment of Patients Treated with Bovine Fibrin as a Hemostatic Agent

1996 ◽  
Vol 76 (06) ◽  
pp. 0925-0931 ◽  
Author(s):  
John F Carroll ◽  
Keith A Moskowitz ◽  
Niloo M Edwards ◽  
Thomas J Hickey ◽  
Eric A Rose ◽  
...  
Keyword(s):  
Coagulation Factors ◽  
Allergic Reactions ◽  
Factor V ◽  
Normal Range ◽  
Serum Samples ◽  
Human Fibrinogen ◽  
Bovine Thrombin ◽  
Thrombotic Complications ◽  
The Mean ◽  
Clinically Significant

SummaryTwenty-one cardiothoracic surgical patients have been treated with fibrin as a topical hemostatic/sealing agent, prepared from bovine fibrinogen clotted with bovine thrombin. Serum samples have been collected before treatment with fibrin and postoperatively between 1 and 9 days, 3 and 12 weeks, and 6 and 8 months. The titers of anti-bovine fibrinogen antibodies, measured by ELISA specific for immunoglobulins IgG or IgM, increased to maximal values after about 8 or 6 weeks, respectively. After 8 months, IgG titers were on average 20-fold lower than the mean maximal value, while IgM titers returned to the normal range. IgG was the predominant anti-bovine fibrinogen immunoglobulin as documented by ELISA, affinity chromatography and electrophoresis. Anti-bovine fibrinogen antibodies present in patients reacted readily with bovine fibrinogen, but did not cross-react with human fibrinogen as measured by ELISA or by immunoelectrophoresis. A significant amount of antibodies against bovine thrombin and factor V has been found, many cross-reacting with the human counterparts. No hemorrhagic or thrombotic complications, or clinically significant allergic reactions, occurred in any patient, in spite of antibody presence against some bovine and human coagulation factors. The treatment of patients with bovine fibrin, without induction of immunologic response against human fibrinogen, appeared to be an effective topical hemostatic/sealing measure.

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