ABSTRACT
OBJECTIVES@#To set up ELISA for detection of atrazine with high precision.@*METHODS@#The reaction condition of indirect-ELISA was optimized, including atrazine-ovalbumin(AT-OVA) concentration and primary antibody concentration, organic solvent, goat anti-rat immunoglobin G-horseradish peroxidase(IgG-HRP) concentration. The actual samples were detected by the ELISA method established in our laboratory. Then the ELISA method was compared with the HPLC.@*RESULTS@#The specification curve of indirect-ELISA was set up after optimization. The relation coefficient R=0.9958. The limit of detection (LOD) was 1.972 ng/ml. The percent recovery of the actual samples was in range of 80%~120%. The ELISA detection sensitivity was higher than the HPLC in the range of 0 ng/ml~6 ng/ml atrazine.@*CONCLUSIONS@#The ELISA to detect atrazine has good specificity and high precision. And it can be applied in testing real atrazine samples replacing of the large-scale instrument.
Subject(s)
Animals , Atrazine , Chromatography, High Pressure Liquid , Enzyme-Linked Immunosorbent Assay , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol (CL) detection.</p><p><b>METHODS</b>The affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by (NH4)2SO4 salt-out method and further by affinity column. At last, the CL detection standard curve which based on indirect competition ELISA was established.</p><p><b>RESULTS</b>The ELISA experiment showed that the antibody titer was 10(6) and the monoclonal antibody affinity constants was 2.90 x 10(10) L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect competition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml.</p><p><b>CONCLUSION</b>The standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced development of the CL immune test paper and CL ELISA kit.</p>
Subject(s)
Animals , Rats , Antibodies, Monoclonal , Chemistry , Antibody Affinity , Clenbuterol , Allergy and Immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay , Limit of DetectionABSTRACT
<p><b>AIM</b>To obtain Clenbuterol monoclonal antibodies.</p><p><b>METHODS</b>Clenbuterol complete antigen was prepared with diazotization method. BALB/c mice was immunized with subtractive immunization, Clenbuterol monoclonal antibody was prepared with rule hybridoma technique.</p><p><b>RESULTS</b>The mice obtained tolerance to BSA by subtractive immunization. The rate of the hybridoma cell with positive reaction which had obtained was 8.2%, and the specific clenbuterol monoclonal antibody was obtained at last.</p><p><b>CONCLUSION</b>Monoclonal antibodies to micromolecule contaminant be prepared by subtractive immunization, could decrease the workload in the bolting of monoclonal antibodies, and increase the chance to obtain the antibody of expected.</p>
Subject(s)
Animals , Female , Male , Mice , Antibodies, Monoclonal , Allergy and Immunology , Clenbuterol , Allergy and Immunology , Hybridomas , Metabolism , Immunization , Methods , Mice, Inbred BALB C , Serum Albumin, Bovine , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>In order to explore the existence of SARS coronavirus (Co-V) and/or its RNA in sewage of hospitals administered SARS patients.</p><p><b>METHODS</b>A novel electropositive filter was used to concentrate the SARS-CoV from the sewage of two hospitals administered SARS patients in Beijing, including twelve 2,500 ml sewage samples from the hospitals before disinfection, and ten 25,000 ml samples after disinfection; as well as cell culture, RT-PCR and sequencing of gene to detect and identify the viruses from sewage.</p><p><b>RESULTS</b>There was no live SARS-CoV detected in the sewage in this study. The nucleic acid of SARS-CoV had been found in the 12 sewage samples before disinfection from both hospitals by semi-nested PCR. After disinfection, SARS-CoV RNA could only be detected from the samples from the 309th Hospital, and the others were negative.</p><p><b>CONCLUSION</b>It provides evidence that there is no live SARS-Cov in the sewage from hospitals with SARS patients though SARS-CoV RNA can be detected.</p>