الفهرس 
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Abstract
Filariasis is a disease caused by filarial parasitic worms, which are microscopic roundworms that live in the blood and tissues of humans. The most critical form of lymphatic filariasis is elephantiasis, which is a serious complication of this disease. The lymphatic form of filariasis will be the focus of this study.
Gold nanoparticles (GNPs) are used in immunochemical studies for identification of protein molecules, as they exhibit unique physicochemical properties allowing easy surface modification and use in different biomedical applications.
This study was carried for evaluation of the diagnostic performance of a novel nano diagnostic assay [nano gold beads based ELISA] in serodiagnosis of lymphatic filariasis and improving identification methods based on detection of circulating filarial antigen.
This study was conducted on 142 individuals.They were divided into 3 group; group I: 78 filarial diseased patients, group II: 54 patients harboring other parasites than lymphatic filariasis and group III: 10 apparently healthy individuals negative control.
Fresh blood samples were collected at daytime from all subjects under study one hour after taking a single dose of 100 mg DEC (Hetrazan) for adults and 50 mg for children as a provocative test.
These samples were examined microscopically for detection of microfilariae by thick bood film stained with Geimsa stain.
Venous blood samples approximately 4 ml were taken from all individuals in EDTA tubes. Sera were separated by centrifugation at 2000 g for 10 min and then fractionated into small epindorfs and stored at -20°C until used.
Production of filarial antigen and its purification was done by ion exchange chromatography on Sephadex A-50 followed by gel filtration chromatography on Sephacryl-S-200 high resolution (HR) column .The eluted proteins from gel filtration column chromatography was analyzed by 12.5% SDS-PAGE under reducing condition and stained with Coomassie blue staine. It showed 2 bands at 18 kilodalton (kDa) and 66.6 kDa which representing purified protein antigen.
Seraria surface antigen was used with complete and incomplete Freund’s adjuvants for immunization of rabbits for preparation of anti- filarial IgG polyclonal antibodies.
Purification of the prepared polyclonal antibodies was carried out by two different methods including 50% ammonium sulphate precipitation method followed by 7% caprylic acid purification method.
The protein content was measured after each purification method; the total protein content of crude rabbits sera was 11.5 mg/ml. After 50% ammonium sulfate precipitation method, the protein content was 5.8 mg/ml, while following 7% caprylic acid precipitation method it dropped to 3.9 mg/ml.
In this work after purification of the polyclonal antibodies. Part of the polyclonal antibodies was conjugated with gold nanoparticles.
Standardization of sandwich ELISA was carried out before the application of the technique on human sera to determine the optimum concentration of coating and conjugate antibody in both types of ELISA.