Tissue transglutaminase antibodies in celiac disease, comparison of an enzyme linked immunosorbent assay and a dot blot assay

ABSTRACT The role of immunoglobulin A (IgA) anti-tissue transglutaminase antibodies (IgA-tTG) as predictors of untreated celiac disease (CoD) is well documented, and the presence and levels of these antibodies are most accurately monitored with native or recombinant human antigens. However, IgA-deficient CoD patients are not identified by IgA serology, and conflicting results concerning the diagnostic validity of IgG antibodies against gliadin (IgG-AGA), endomysium (IgG-EmA), and tTG (IgG-tTG) have been reported. The aim of the present study was to evaluate the utility of IgG-tTG for the detection of CoD in IgA-deficient patients. Samples from 115 IgA-deficient and 200 IgA-sufficient subjects were collected and tested for the presence of IgA and IgG antibodies against tTG, EmA, and AGA. Antibodies against tTG were measured by an enzyme-linked immunosorbent assay based on recombinant human tTG, and antibodies against EmA were determined by immunofluorescence. The values for IgG-tTG showed a higher correlation (correlation coefficient [r] = 0.91) with those for IgG-EmA for the IgA-deficient subjects than for the IgA-sufficient subjects (r = 0.88). The overall concordance of the positive and negative results between IgG-tTG and IgG-EmA was 97%, and the IgG-tTG assay discriminated between IgG-EmA-positive and -negative subjects with IgA deficiency at a rate of 100%. Elevated levels of IgG-tTG and IgG-EmA were measured in 70% of the IgA-sufficient subjects. IgG-tTG detection with recombinant human tTG is a good alternative to IgG-EmA detection, and the addition of IgG-tTG assessment to present screening methods may improve the ability to identify IgA-deficient subjects with CoD.

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Development of an Enzyme-Linked Immunosorbent Assay Using a Recombinant LigA Fragment Comprising Repeat Domains 4 to 7.5 as an Antigen for Diagnosis of Equine Leptospirosis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00245-13 ◽

2013 ◽

Vol 20 (8) ◽

pp. 1143-1149 ◽

Cited By ~ 10

Author(s):

Weiwei Yan ◽

Muhammad Hassan Saleem ◽

Patrick McDonough ◽

Sean P. McDonough ◽

Thomas J. Divers ◽

...

Keyword(s):

Immunosorbent Assay ◽

Indirect Elisa ◽

Enzyme Linked Immunosorbent Assay ◽

Dot Blot ◽

Serum Dilution ◽

Content Type ◽

Diagnostic Antigen ◽

Repeat Domain ◽

Associated Proteins ◽

Dot Blot Assay

ABSTRACTLeptospiraimmunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5–5.5), 5.5th to 6.5th (LigACon5.5–6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that LigACon4-7.5 is the best candidate diagnostic antigen in a slot-shaped dot blot assay. LigACon4-7.5 was further evaluated as an indirect enzyme-linked immunosorbent assay (ELISA) antigen for the detection ofLeptospiraantibodies in equine sera. This assay was evaluated with equine sera (n= 60) that were microscopic agglutination test (MAT) negative and sera (n= 220) that were MAT positive to the 5 serovars that most commonly cause equine leptospirosis. The indirect ELISA results showed that at a single serum dilution of 1:250, the sensitivity and specificity of ELISA were 80.0% and 87.2%, respectively, compared to those of MAT. In conclusion, an indirect ELISA was developed utilizing a recombinant LigA fragment comprising the 4th to 7.5th repeat domain (LigACon4-7.5) as a diagnostic antigen for equine leptospirosis. This ELISA was found to be sensitive and specific, and it yielded results that concurred with those of the standard MAT.

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Tissue transglutaminase autoantibody enzyme-linked immunosorbent assay in detecting celiac disease

Gastroenterology ◽

10.1016/s0016-5085(98)70008-3 ◽

1998 ◽

Vol 115 (6) ◽

pp. 1322-1328 ◽

Cited By ~ 375

Author(s):

Satu Sulkanen ◽

Tuula Halttunen ◽

Kaija Laurila ◽

Kaija-Leena Kolho ◽

Ilma R. Korponay-Szabó ◽

...

Keyword(s):

Celiac Disease ◽

Immunosorbent Assay ◽

Tissue Transglutaminase ◽

Enzyme Linked Immunosorbent Assay

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Detection of M2 antimitochondrial antibodies by dot blot assay is more specific than by enzyme linked immunosorbent assay

Pathologie Biologie ◽

10.1016/j.patbio.2007.05.001 ◽

2008 ◽

Vol 56 (1) ◽

pp. 10-14 ◽

Cited By ~ 6

Author(s):

I. Bargou ◽

A. Mankaï ◽

A. Jamaa ◽

I. Ben Jazia ◽

K. Skandrani ◽

...

Keyword(s):

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Antimitochondrial Antibodies ◽

Dot Blot ◽

Dot Blot Assay

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Chemiluminescent Immunoassay versus Enzyme Linked Immunosorbent Assays for IgA Anti-Tissue Transglutaminase Antibodies Assessment in Celiac Disease Children

Revista de Chimie ◽

10.37358/rc.20.2.7890 ◽

2020 ◽

Vol 71 (2) ◽

pp. 45-51

Author(s):

Oana Belei ◽

Emil Radu Iacob ◽

Daniela Iacob ◽

Elena Amaricai ◽

Otilia Marginean

Keyword(s):

Celiac Disease ◽

Tissue Transglutaminase ◽

Immunoglobulin A ◽

Hla Typing ◽

Intestinal Biopsy ◽

Enzyme Linked Immunosorbent Assay ◽

Digestive Endoscopy ◽

Statistical Comparison ◽

Tissue Transglutaminase Antibodies ◽

Chemiluminescent Immunoassay

Recent describing of atypical, silent and latent form of celiac disease (CD) increased the preocupation for screening methods.To perform a comparative study of immunoglobulin A (IgA) anti tissue-transglutaminase (tTG) antibodies (Ab) assessment using chemiluminescence versus Enzyme Linked Immunosorbent Assay (ELISA).The study included two lots.The first lot consisted in 35 biopsy confirmed CD children aged between two and 18 years.The control lot included 40 children with normal duodenal morphology on intestinal biopsy that underwent upper digestive endoscopy for different gastrointestinal symptoms.Serum samples were provided from all subjects for IgA measurement, IgA anti tTG and EMA assessment. Immulite 2500 Anti tTG IgA (Siemens Co, LA, USA) and ImmuLisa anti-hu tTG antibody IgA ELISA (Immco Diagnostics Inc, NY, USA) kits were used for tTG-Ab assessment. All children underwent HLA typing for DQ2/DQ8. The sensitivity for IgA tTG assessment was greater for chemiluminescence (93,3%) versus ELISA (86,6%), p[0,05, while specificity, positive and negative predictive value didn�t register significant differences. Statistical comparison of the two tested methods revealed a better sensitivity for chemiluminescence. Diagnosis algorithm optimisation may be obtained by associating IgA anti tTG-Ab assessment using chemiluminescence followed by EMA confirmation by indirect immunofluorescence for CD screening.

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Evaluation of the INOVA Diagnostics Enzyme-Linked Immunosorbent Assay Kits for Measuring Serum Immunoglobulin G (IgG) and IgA to Deamidated Gliadin Peptides

Clinical and Vaccine Immunology ◽

10.1128/cvi.13.1.150-151.2006 ◽

2006 ◽

Vol 13 (1) ◽

pp. 150-151 ◽

Cited By ~ 44

Author(s):

Harry E. Prince

Keyword(s):

Celiac Disease ◽

Immunosorbent Assay ◽

Immunoglobulin G ◽

Immunoglobulin A ◽

Serum Immunoglobulin ◽

Enzyme Linked Immunosorbent Assay ◽

Gliadin Antibodies ◽

Gliadin Peptides ◽

Disease Specific ◽

Inova Diagnostics

ABSTRACT New assays for antibodies to deamidated gliadin peptides (DGP) expressing celiac disease-specific epitopes were evaluated using 154 sera previously tested for endomysial immunoglobulin A (IgA) (EMA), transglutaminase IgA (TGA), and conventional gliadin antibodies. DGP antibody results showed 97% concordance with EMA and TGA results. Of 56 sera negative for EMA and TGA but positive for conventional gliadin antibodies, 54 (96%) were negative for DGP antibodies.

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Relationships between Clinical Presentation, Serology, Histology, and Duodenal Deposits of Tissue Transglutaminase Antibodies in Pediatric Celiac Disease

Digestive Diseases ◽

10.1159/000490377 ◽

2018 ◽

Vol 36 (5) ◽

pp. 369-376 ◽

Cited By ~ 1

Author(s):

Nurit Loberman-Nachum ◽

Michael Schvimer ◽

Camila Avivi ◽

Iris Barshack ◽

Avishay Lahad ◽

...

Keyword(s):

Celiac Disease ◽

Immunohistochemical Staining ◽

Clinical Presentation ◽

Tissue Transglutaminase ◽

Mucosal Damage ◽

High Serum ◽

Tissue Transglutaminase Antibodies ◽

Antibody Levels ◽

Age Range ◽

Multiple Symptoms

Background: The clinical, histological, and serological spectrum of celiac disease (CD) vary widely. We aimed to examine relationships between symptoms, serum anti-tissue transglutaminase antibodies (tTG) levels, mucosal damage, and mucosal anti-tTG deposits in pediatric CD. Methods: A retrospective single-center, cohort study of children referred for endoscopy with suspected CD during 2011–2014. We retrieved the clinical data, blindly reviewed duodenal biopsies, and performed immunohistochemical staining for anti-tTG deposits. Patients were classified as monosymptomatic or polysymptomatic. Mucosal anti-tTG deposits were classified according to the location of deposits, dominant intensity, maximal intensity, and percentage of stained area. Results: Of 252 patients with confirmed CD, complete data were available for 100: 37 males in the age range 1.3–16.7 with median 4.0 years. Monosymptomatic patients (n = 54) presented at an older age than polysymptomatic patients (1.3–15.5, median 8.1 vs. 1.3–16.7, median 6.3 years, p = 0.026). Marsh 2–3c was more prevalent in polysymptomatic patients (93 vs. 78%, p = 0.028). The intensity of mucosal anti-tTG deposits correlated with serum anti-tTG levels but not with the clinical presentation. Conclusions: Multiple symptoms and high serum anti-tTG antibody levels correlated with mucosal damage in children with CD. The role of immunohistochemical staining for intestinal anti-tTG mucosal deposits in the diagnosis of borderline CD is not yet established.

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