A Rapid Lateral Flow Immunoassay for the Detection of Tyrosine Phosphatase-Like Protein IA-2 Autoantibodies in Human Serum

Kikkas, Ingrid; Mallone, Roberto; Larger, Étienne; Volland, Hervé; Morel, Nathalie

doi:10.1371/journal.pone.0103088

Cited by 13 publications

(9 citation statements)

References 23 publications

(19 reference statements)

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“…At least one of these autoantibodies is present in 95% of the individuals who have DT1, after the detection of the hyperglycemia. These auto-antibodies can be effectively used for the prediction of type 1 diabetes, and they can serve as early markers of DT1, given their persistence in patients' sera for years prior to the development of diabetes type 1, at the time of diagnosis and even after diagnosis 3 , 4 .…”

Section: Introductionmentioning

confidence: 99%

Research of anti-GAD and anti-IA2 autoantibodies by ELISA test in a series of Moroccan pediatric patients with diabetes type 1

et al. 2020

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Introduction: Type I diabetes (T1D) is an autoimmune disease with a prediabetic, asymptomatic period characterized by the selective destruction of insulin-producing β cells. During the pre-clinical phase, various auto-antibodies are generated against several beta cell antigens such as anti glutamate acid decarboxylase (Anti-GAD), anti tyrosine phosphatase (Anti-IA2). Today, the coupled detection of Anti-IA2 with that of Anti-GAD proves its great importance in the diagnosis and prediction of type 1 diabetes. The combined positivity for both antibodies has a specificity and a positive predictive value of 100%. Objectives: In this work, we evaluate the diagnostic value of anti-GAD and anti-IA2 antibodies in a series based on 78 Moroccan subjects initially under 16, suspected T1D. Results and Discussion: Our series consists mainly of 74% of newly diagnosed patients for T1D and 26% of confirmed diagnostic patients, of whom 52% are females. The mean age of diagnosis is 7 ± 4 years, the mean of HbA1c at the time of diagnosis is 11.63 ± 2.16%, and the percentage of family history in our series is 69%. The proportion of positive results for anti-IA2 antibodies and anti-GAD antibodies are, respectively, 76.92% and 62.82%, and 52.56% of patients are positive for both auto-antibodies. This study confirms that anti-GAD and anti-IA2 auto-antibodies assays can detect patients early and the autoantibodies can persist several years after diagnosis of type 1 diabetes. Conclusion: This study confirmed the diagnosis and classification of T1D (type 1A) in 87.18% of patients, and we reported that the prevalence of anti-GAD and anti-IA2 is higher in girls than in boys. Keywords: Type 1 diabetes; autoimmunity; autoantibodies; anti-GAD; anti-IA2; ELISA; classification.

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Section: Introductionmentioning

confidence: 99%

Research of anti-GAD and anti-IA2 autoantibodies by ELISA test in a series of Moroccan pediatric patients with diabetes type 1

et al. 2020

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“…In the proposed format, the detected complexes are formed after the sample has rapidly moved along the tracks on the working membrane between neighboring zones, which are only 2 mm apart. Several immunochromatographic tests that exclude the conjugate pad have been reported [13][14][15]. In these assays, the conjugate is preincubated with the sample beyond the test strip.…”

Section: Discussionmentioning

confidence: 99%

Development of a Lateral Flow Highway: Ultra-Rapid Multitracking Immunosensor for Cardiac Markers

Byzova

Vengerov

Voloshchuk

et al. 2019

Sensors

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The integration of several controlled parameters within a single test system is experiencing increased demand. However, multiplexed test systems typically require complex manufacturing. Here, we describe a multiplexed immunochromatographic assay that incorporates a conventional nitrocellulose membrane, which is used together with microspot printing, to construct adjacent microfluidic "tracks" for multiplexed detection. The 1 mm distance between tracks allows for the detection of up to four different analytes. The following reagents are applied in separate zones: (a) gold nanoparticle conjugates with antibodies against each analyte, (b) other antibodies against each analyte, and (c) antispecies antibodies. The immersion of the test strip in the sample initiates the lateral flow, during which reagents of different specificities move along their tracks without track erosion or reagent mixing. An essential advantage of the proposed assay is its extreme rapidity (1-1.5 min compared with 10 min for common test strips). This assay format was applied to the detection of cardiac and inflammatory markers (myoglobin, D-dimer, and C-reactive protein) in human blood, and was characterized by high reproducibility (8%-15% coefficient of variation) with stored working ranges of conventional tests. The universal character of the proposed approach will facilitate its use for various analytes.

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“…According to literature [74], the cysteine residues were unlikely to form disulfide bonds, contributing little to the three dimensional structure of the protein. Probably, disulfide bridges are formed later due to the action of reactive oxygen species during storage or the IA-2 ic PTP and juxtamembrane domains tendency to dimerize [75–77], leading to polymerization. This in vitro observation could be a resemblance of the in vivo regulation of many receptor-type PTPs.…”

Section: Discussionmentioning

confidence: 99%

“…The parameters of sensitivity (58.3 %), relative sensitivity (59.5 %) and specificity (94.9 %), although lower than those for RBA, are more than acceptable for use as a first line screening method for assessing humoral markers. It is difficult to compare the performance of this assay with those of previous studies where IA-2A is detected because in those studies different antigens were used, as well as different patient populations and healthy control individuals (with variables such as number, ethnicity and, possibly, sample matrix) [62, 63, 74–77]. The lower sensitivity showed by CL- b ELISA, in comparison with RBA, could be a consequence of partial denaturation of TrxIA-2 ic when adsorbed onto the surface of polystyrene microplates.…”

Section: Discussionmentioning

confidence: 99%

Novel prokaryotic expression of thioredoxin-fused insulinoma associated protein tyrosine phosphatase 2 (IA-2), its characterization and immunodiagnostic application

et al. 2016

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BackgroundThe insulinoma associated protein tyrosine phosphatase 2 (IA-2) is one of the immunodominant autoantigens involved in the autoimmune attack to the beta-cell in Type 1 Diabetes Mellitus. In this work we have developed a complete and original process for the production and recovery of the properly folded intracellular domain of IA-2 fused to thioredoxin (TrxIA-2ic) in Escherichia coli GI698 and GI724 strains. We have also carried out the biochemical and immunochemical characterization of TrxIA-2icand design variants of non-radiometric immunoassays for the efficient detection of IA-2 autoantibodies (IA-2A).ResultsThe main findings can be summarized in the following statements: i) TrxIA-2ic expression after 3 h of induction on GI724 strain yielded ≈ 10 mg of highly pure TrxIA-2ic/L of culture medium by a single step purification by affinity chromatography, ii) the molecular weight of TrxIA-2ic (55,358 Da) could be estimated by SDS-PAGE, size exclusion chromatography and mass spectrometry, iii) TrxIA-2ic was properly identified by western blot and mass spectrometric analysis of proteolytic digestions (63.25 % total coverage), iv) excellent immunochemical behavior of properly folded full TrxIA-2ic was legitimized by inhibition or displacement of [35S]IA-2 binding from IA-2A present in Argentinian Type 1 Diabetic patients, v) great stability over time was found under proper storage conditions and vi) low cost and environmentally harmless ELISA methods for IA-2A assessment were developed, with colorimetric or chemiluminescent detection.Conclusions E. coli GI724 strain emerged as a handy source of recombinant IA-2ic, achieving high levels of expression as a thioredoxin fusion protein, adequately validated and applicable to the development of innovative and cost-effective immunoassays for IA-2A detection in most laboratories.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0309-2) contains supplementary material, which is available to authorized users.

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A Rapid Lateral Flow Immunoassay for the Detection of Tyrosine Phosphatase-Like Protein IA-2 Autoantibodies in Human Serum

Cited by 13 publications

References 23 publications

Research of anti-GAD and anti-IA2 autoantibodies by ELISA test in a series of Moroccan pediatric patients with diabetes type 1

Research of anti-GAD and anti-IA2 autoantibodies by ELISA test in a series of Moroccan pediatric patients with diabetes type 1

Development of a Lateral Flow Highway: Ultra-Rapid Multitracking Immunosensor for Cardiac Markers

Novel prokaryotic expression of thioredoxin-fused insulinoma associated protein tyrosine phosphatase 2 (IA-2), its characterization and immunodiagnostic application

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