PENGEMBANGAN KIT DIAGNOSTIK ANTI-HBSAG KUANTITATIF BERBASIS ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

• Main Author: Khairunnisa
• Format: Final Project
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/76400
• Institution: Institut Teknologi Bandung

Hepatitis is an inflammatory condition of the liver that can be caused by viral infections, heavy alcohol consumption, autoimmunity, drugs, and toxins. The Data and Information Center of the Indonesian Ministry of Health in 2014 stated that Indonesia has the second highest Hepatitis B endemic condition in Southeast Asian countries after Myanmar. This is contradicted by the data of Hepatitis B immunization (HB0) in Indonesia in 2007-2017 which is always above 90%. This contradictory condition should be the focus to evaluate the success of Hepatitis B vaccination in Indonesia. Unfortunately, this extensive immunization coverage in Indonesia is not formally followed by an evaluation of its vaccination success allegedly because the anti-HBsAg diagnostic kit used is relatively expensive and not widely available. Based on this, research activities were carried out to develop an ELISA-based anti-HBsAg diagnostic kit that can detect anti-HBsAg titers in blood samples quantitatively. Scope of study of this research is determination of the diagnostic kit format, washing solution, incubation method and load capture substance. Testing kit formats that use capture antigens and anti-human IgG-HRP mouse antibody conjugates results in nonspecific binding (false positive) and washing using Glycine-Tween, Tris-HCl, and TBS washing solutions is not effective in removing these nonspecific bindings. further research this study states that nonspecific binding can be eliminated by changing the format of the anti-HBsAg diagnostic kit. Load capture substance test (25, 50, 100 and 500 ng / well) did not have a significant effect on the performance of this anti-HBsAg diagnostic kit. Furthermore, from this study it was also found that testing by first incubating the sample and conjugate in the tube gave better examination results compared to incubating the sample directly in the ELISA well and this method only requires 1 washing process, so it can save operator time and labor. Overall, this study has successfully developed an anti-HBsAg diagnostic kit that can detect anti-HBsAg antibody titers well without causing nonspecific binding.