Recombinant Protein Production from TPO Gen Cloning and Expression for Early Detection of Autoimmune Thyroid Diseases

Aulanni’am, Aulanni’am; Wuragil, Dyah Kinasih; Soeatmadji, Djoko Wahono; Zulkarnain, Zulkarnain; Marhendra, Agung Pramana Warih

doi:10.1088/1757-899x/299/1/012013

Cited by 1 publication

(1 citation statement)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The general overview of rapid tests is generated based on the concept in Figure 1 . The details of our manual protocol for this work were arranged by our previous work [ 22 ]. To obtain the best result, the kit was incubated at room temperature for a while to allow the sample and buffer to mix gently.…”

Section: Methodsmentioning

confidence: 99%

The early detection of type 1 diabetes mellitus and latent autoimmune diabetes in adults (LADA) through rapid test reverse-flow immunochromatography for glutamic acid decarboxylase 65 kDa (GAD65)

Aulanni’am

Wuragil

Susanto

et al. 2022

Heliyon

Self Cite

View full text Add to dashboard Cite

Background: Diabetes mellitus (DM) is a chronic and costly disease that has become a primary concern worldwide. Type 1 diabetes mellitus is categorized as an autoimmune disease, which results in islet cell apoptosis and insulindependent. GAD 65 is known as a potential marker of impaired pancreatic β cell function that appears in the initial phase of type 1 DM and latent autoimmune diabetes in adults (LADA). This study aimed to develop a novel rapid test of anti-GAD 65 autoantibodies in human serum samples. Methods: We have developed a rapid test for anti-GAD65 autoantibodies in this assay based on the reverse-flow immunochromatography method. Human recombinant-protein antigen for GAD 65 was attached as the control line over the nitrocellulose membrane. On the other side, the goat anti-mouse immunoglobulin G (IgG) was coated on the same membrane as a control line. The positive result for GAD 65 was confirmed by a colloidal gold signal on the strip. Our novel assay analyzed 276 healthy subjects and 51 type 1 diabetes individuals serum samples from several ethnicities in Indonesia for this study. Results: Among the 276 healthy samples, 225 samples were identified as positive for anti-GAD 65 autoantibodies, while 51 samples were negative. Interestingly, the positive results for anti-GAD 65 autoantibodies were linear to the decreasing of high-density lipoprotein (HDL) levels and inversely associated with triglyceride levels. A significant correlation with age was observed in all groups. The sensitivity and specificity test proved that this kit has higher accuracy (AUC value ¼ 0.960). Conclusion:The significant advantages of our rapid test for anti-GAD 65 autoantibodies provide higher sensitivity, specificity, and stability compared to previous commercial kits. Therefore, it could be proposed as the future clinical diagnostic kit for patient management of type 1 DM.

show abstract

Section: Methodsmentioning

confidence: 99%