The first one is specific monoclonal antibody used in coating of ELISA plate, while the other one is laboratory prepared reference hyper immune sera of the other host
The first one is specific monoclonal antibody used in coating of ELISA plate, while the other one is laboratory prepared reference hyper immune sera of the other host. animals. This parasite constitutes a native health problem in Egypt because of its frequent occurrence and was therefore selected for the study. The development of suitable sustainable strategies for accurate early diagnosis is essential for controlling contamination. Diagnosis is usually primitively based on identifying the parasites eggs in the fecal samples. However, eggs can generally be detected after the maturation of the worms when they begin egg shedding (15-18 weeks post contamination). Rapid diagnostic methods would facilitate control of the parasite. Immunoassays as diagnostic tools can be employed either by antibody detection in the infected serum (1) or by antigen detection GPI-1046 (2). The time required in the former 2 to 4 weeks p.i. whereas in the latter, 4 to 6 6 weeks p.i. Coproantigens are antigens prepared from fecal sample of infected animal and have shown to be effective for the diagnosis of GPI-1046 a wide range of human and animal intestinal infections. In theory, antibodies raised against whole parasite extracts are coated on microtiter plates, and subsequently fecal antigen is usually captured and detected with the same or second parasite-specific antibody in a capture assay(3, 4). A diagnostic antigen in the bile and feces from infected cattle was detected and characterized using Enzyme-Linked Immunotransfer Blot technique (EITB) (5). Coproantigen ELISA was developed for the detection of excretory secretory antigens in humans with fascioliasis (6). To monitor the Rabbit Polyclonal to ACTN1 efficacy of treatment, the detection of antigen rather than antibody is considered more reliable for diagnosing contamination. In the present study, we statement the first time of appearance and dynamics of coproantigens in feces and circulating antibodies in sera in rabbits experimentally infected with different doses of EMC. Material & Methods Animal contamination Twelve parasite-free New Zealand white rabbits about 2 GPI-1046 kg b.wt. were used in the experiment. Nine rabbits were orally infected with 10, 25 and 30 EMC (10 days aged); (3 rabbits for each dose level) while the remaining 3 rabbits served as uninfected controls. Fecal and blood samples (for separation of serum) were collected immediately before contamination and weekly for successively 12 weeks and stored at -20C. Thereafter, the rabbits were sacrificed and flukes were collected after maceration of the liver using scalpel and needle in warm water. Rabbit hyper immune sera were raised separately against coproantigens, ES antigen and egg antigen (7). Excretory-Secretory (ES) antigen preparation ES antigens were prepared from (8) with slight modifications. Adult worms GPI-1046 obtained from bovine livers were washed repeatedly (3-5 occasions) in M PBS, incubated for 3 h at GPI-1046 37 C (one worm/5ml in 0.01 M PBS, pH 7.4). After incubation, the worms were removed and the supernatant fluid (PBS + ES) was collected and subjected to high speed centrifugation (12000 rpm) for 1 h at 4 C. The supernatant was separated and designated as ES antigen. The protein content was measured (9). The antigen was aliquoted and stored at -70 C until use. Fasciola gigantica Coproantigen preparation Fecal supernatants were processed for coproantigen immune detection according to El-Bahy et al. (1992). Five grams of sample was mixed separately in an equivalent amount of distilled water and sonicated for 5 minutes under 150 watt interrupted pulse output at 50% power cycle using a sonifier cell disrupter. The fecal suspensions were centrifuged at 3000 rpm for.