Blastocystis sp. : Evaluation of polyclonal antibody prepared from crude protein for serological diagnosis using Rabbit serum

The diagnosis of Blastocystis infection is still based on the clinical sign which is not specific and there is no available serologic test for it. This study aimed to evaluate the polyclonal antibody prepared form crude protein of Blastocystis for the development of the Blastocystis serological test. Crude protein was extracted from the yeast of Blastocystis sp, then inoculated into rabbits to produce the antibody of crude protein. The serum of rabbits would be collected before and after immunization to compare the antibody titer. The profile of crude protein was analyzed using SDS-Page. The rabbit serum was analyzed using ELISA and Western Blot. The SDS-Page result showed bands in 100 kDa, 90 kDa, 70 kDa, 60 kDa, 58 kDa, 50 kDa, 40 kDa, 35 kDa, 30 kDa and 27 kDa. The ELISA assay showed that there was an increase in antibody titer of crude protein after immunization. Western Blot showed that three proteins (30 kDa, 40 kDa and 50 kDa) having immunogenicity characteristic. It is concluded that protein 30 kDa, 40 kDa and 50 kDa prepared from the crude protein of Blastocystis sp. can be used for developing a serologic test for Blastocystis infection. Keywords: Blastocystis sp, Crude Protein, Polyclonal Antibody  

Characterization and Production of Polyclonal Antibody Anti Excretory Secretory Protein of Blastocystis sp.

This study aims to produce and characterize polyclonal antibody anti excretory secretory (ES) protein of Blastocystis sp. ES protein pro le was analyzed using SDS-PAGE and used to immunize 2 rabbits. Rabbit’s serum were analyzed using indirect ELISA and Western Blot. The result showed that molecular weight of ES protein of Blastocystis sp was 40 and 50 kDa and the protein was immunogenic. Both ES protein and antibody anti ES of Blastocystis sp can be promoted as diagnostic kit. Key words : Blastocystis sp, Excretory Secretory Protein, Polyclonal Antibod

Profil Antigen Dan Antibodi Poliklonal Anti Blastocystis sp Pada Sapi Penelitian Eksploratif Laboratorik

Blastocystis is an intestinal parasite of human and wide range of animals, widely prevalent in many countries. Soluble protein can induce the cellular immunity response in host, meanwhile ESA protein is responsible for the process of inflammation acted as immunomodulator. This study aims to understanding the antigenic profile of soluble and ESA protein, the antibody titer and the immunogenicity. The antigen profile was identified using SDS-Page, while the antibody titer was measured using ELISA and western blotting is performed to understand the immunogenicity of protein. The result of SDS-Page shows that ESA contains 2 proteins (50 kDa and 40 kDa), while Soluble protein consists of 10 proteins (100 kDa, 90 kDa, 70 kDa, 60 kDa, 50 kDa, 40 kDa, 35 kDa, 30 kDa, and 27 kDa). There is also a significance increase of the antibody titer after immunization using Soluble and ESA protein. Western blotting analysis demonstrates that there are 2 proteins (50 kDa and 40 kDa) recognized from ESA protein and 3 proteins (50 kDa, 40 kDa, and 30 kDa) recognized from Soluble protein