RESUMO
To evalue the coincidence and correlation between the four domestic quantity assay reagents and with ARCHITECTi2000 immunoassay system. 185 weak-reactive serum samples and standard materials of different concentrations were tested by four domestic quantity assay reagents for HBsAg test and ARCHITECTi2000 immunoassay system. The coincidence, the precision and the correlations between different systems were analyzed. The coincidence rates of the results of 0.05-1.00 IU/ml samples between the four domestic quantity assay reagents and ARCHITECTi2000 immunoassay system were 25.93%, 35.19%, 51.85% and 18.52% respectively, and for those results of more than 1.00 to 10.00 IU/ml samples the coincidence rates were 71.76%, 87.79%, 95.42% and 69.47% respectively. The samples of 0.05 to 0.80 IU/ml weak-reactive serum samples detected by the i2000 system were all negative detected by the four domestic systems. The coincidence rates of more than 7.93 IU/ml serum samples detected by i2000 system were 100% detected by the four domestic systems. The correlations of the four domestic quantity assays were around 0.8629 to 0.9265. The analysis sensitivity of the four domestic quantity assay reagents were below the i2000 system. The results of under 0.80 IU/ml samples detected by i2000 system were disaccord with the results detected by the four domestic systems, whereas for the sapmples over 7.93 IU/ml the results were consistent.
RESUMO
<p><b>AIM</b>To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres (IMMS) based on enzyme-linked immunosorbent assay (ELISA) for the purification of human albumin from human serum.</p><p><b>METHODS</b>Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide (EDC). Finally rabbit anti-human serum albumin (HSA) antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum.</p><p><b>RESULTS</b>The result of the experiment was that the recovery of human albumin with IMMS was (86 +/- 4)%, and IMMS were reused for two other purifying cycles, the results of which were (69.0 +/- 0.6)% and (40.8 +/- 0.8)%, and the purity of the product was about 90%.</p><p><b>CONCLUSION</b>The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of high-purity HSA.</p>
Assuntos
Humanos , Ensaio de Imunoadsorção Enzimática , Métodos , Separação Imunomagnética , Métodos , Microesferas , Poliestirenos , Química , Reprodutibilidade dos Testes , Albumina Sérica , Alergia e ImunologiaRESUMO
Mice and 3-day-old chickens were orally inoculated with the recombinant attenuated Salmonella typhimurium strain ZJ111 carrying pcDNA3-F expression plasmid encoding the fusion protein of Newcastle disease virus (NDV). The results showed that ZJ111/pcDNA3-F was relatively safe. The recombinant plasmid pcDNA3-F was stable within the host stain ZJ111 in vitro and in vivo as shown by restriction enzyme analysis and PCR identification of the F gene. In an experimental vaccination study, 3-day-old chickens were orally immunized with ZJ111/pcDNA3-F with a dose of 108 cfu per chicken and boosted two weeks later. At week 4 post boosting, all chickens were challenged with a lethal dose of a virulent NDV strain F48 E9. The results showed that oral vaccination with ZJ111/pcDNA3-F induced stronger humoral and cellular immune responses than intramuscular immunization with naked pcDNA3-F plasmid. It also exhibited higher protection rate than the latter (66.7% vs 50%). This study indicates that the DNA vaccine using attenuated Salmonella typhimurium as delivery carrier had good safety, stability and immunogenicity and exhibited good potential of low cost and convenience for poultry disease control.