PENGUJIAN ANTIGENISITAS EPITOP PEPTIDA SINTETIK DARI PROTEIN F HEPATITIS C VIRUS (HCV) DENGAN METODE INDIRECT ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)

• Main Author: Joshua Wiratama, Emilio
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/48720
• Institution: Institut Teknologi Bandung

Hepatitis C is a liver disease caused by the hepatitis C virus (HCV) which could develop into liver cancer. An estimated 71 million people in the world have hepatitis C, 3 million of which are Indonesian people. As of now, no vaccine has been found that could prevent an infection. Available medical treatments for this disease include detection for those who are at a high risk of infection, and prescription of antivirals for those who come up positive. Early treatment is crucial in achieving recovery, which is why a fast and accurate detection kit is needed for this disease. Up until now, Indonesia still uses imported hepatitis C detection kits. The ability to produce hepatitis C detection kit independently becomes an important matter in Indonesia, not only to reduce the price of diagnosis, but also to reduce Indonesia’s dependency on other countries. Previous researchers have developed an epitope from F protein which hypothetically could be used to develop an HCV diagnostic kit, but the antigenicity of said epitope has yet to be studied. In this research, the antigenicity of the developed epitope will be tested with an indirect ELISA method. In this research, serum from a healthy individual who has never been infected with HCV is used as a negative control, and a pooled serum of patients co-infected with HIV and HCV stored in RSUP Hasan Sadikin, West Java, is used to test the antigenicity. A comparison control is done using a commercial ELISA HIV detection kit by Autobio, using the same sera as the negative control and sample. The antigen coating process in this research uses PBS as a coating buffer, and is then continued with incubation at 4oC for 16 hours. The blocking process is done with a solution of 5% skim milk in PBST as a blocking buffer, which is then continued with incubation at 4oC for 24 hours. The ELISA process is done according to the instruction manual of the Autobio kit. The test is done with a pooled serum of HCV and HIV co-infected patients with and without 1:1 dilution, and an epitope concentration of 25 ?g/mL to 0,1 ?g/mL. The results of this research shows that antigenicity is observed even on 0,1 ?g/mL epitope concentration and a 1:1 diluted serum. The results indicate that the developed synthetic epitope from the F protein of HCV has an observed antigenicity towards sera of HCV infected Indonesian patients. Further research is needed to define the detection limit, sensitivity, and specificity of the epitope.