Karakterisasi dan Isolasi Protein Spesifik dari Material Excretory-Secretory (ES) Toxocara cati untuk Pengembangan Diagnostik Toxocariasis dengan Teknik ELISA
Abstract: The objective of
this study was to get the specific protein of T. cati that could be used as
biomarker in kit diagnostic to toxocariasis diagnosis through antibody test by
ELISA technique. This study was conducted by isolating and characterizis T.
cati protein from secretory-excretory (ES) material of L2 dormant (L2D) by pure
protein that was reactived with polyclonal antibody sera. For the first step,
ES protein of L2D was identified by SDS-PAGE with coumsie blue staining. The
second step, protein was transferred to nitrocellulose membrane by using
polyclonal antibody of anti-L2D T. cati that was visualized through goat-anti
rabbit conjugate and BCIP+NBT staining. The third step to decide the specific
protein fractions based on the molecule weight (MW), and then the specific
protein was isolated by elution technique. The fourth was to evaluate the
antigenicity, sensitivity and specificity of pure protein by IgG-ELISA as a
response against L2D T. cati immunization toward mice. The results of the study
showed that: T. cati have more protein with varied molecular weight ranging
from 8 to 200 kDa. The specific proteins of T. cati recognized by the anti-L2D
T. cati sera are the proteins of MW 168, 140, 120, 80, 70, 42, 32, 30, 24, 18,
14, 12 dan 10 kDa. The antigenicity of protein of T. cati was higher than that
of other worms (D_caninum and Ancylostoma spp.). The sensitivity of protein of
T. cati in mice with immunization was 100%, the specificity was 87.5% and the
false positive was 12.5%.
Penulis: Kusnoto, Sri Subekti,
I Ketut Sudiana, Soedarto
Kode Jurnal: jpbiologidd110044
