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All Journal Makara Journal of Science
Wuragil, Dyah Kinasih
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Development of an Indirect ELISA Kit for Measurement of Circulating Thyroid Peroxidase Levels in Patients with Hypothyroidism Aulanni’am, Aulanni’am; Ramadani, Devi; Khairana, Almas Dwi; Fiddaroini, Saidun; Nur, Muhammad Fikri; Wijaya, Andreas Budi; Rosandi, Rulli; Wuragil, Dyah Kinasih; Riawan, Wibi
Makara Journal of Science Vol. 30, No. 3
Publisher : UI Scholars Hub

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Abstract

Hypothyroidism, characterized by insufficient thyroid hormone production, affects approximately 5% of the population, especially women, thus increasing pregnancy-related risks. Conventional screening mainly measures thyroid-stimulating hormone (TSH) and free thyroxine (FT4). Detecting the presence of anti-thyroid peroxidase (anti-TPO) antibodies indi-cates autoimmune thyroid disease. This study developed an indirect enzyme-linked immunosorbent assay (ELISA) kit for detecting circulating TPO using polyclonal antibodies generated in rabbits. TPO was isolated from positive sera of pa-tients with hypothyroidism, purified by electroelution, and confirmed as a 52-kDa protein by Western blot. Rabbits immunized with this protein produced polyclonal antibodies with the highest titer (OD = 0.334) at week 8. Dot blot analysis demonstrated a detection capability up to a serum dilution of 1:80. The developed indirect ELISA showed reliable sensi-tivity and indicates potential cost-effectiveness. These findings demonstrate its potential as a diagnostic tool for early hypothyroidism detection and monitoring, providing a foundation for further clinical validation and broader application.
Co-Authors Andreas Budi Wijaya Aulanni’am, Aulanni’am Fiddaroini, Saidun Khairana, Almas Dwi Nur, Muhammad Fikri Ramadani, Devi Rulli Rosandi Wibi Riawan