A competitive indirect time-resolved fluoroimmunoassay(TRFIA) was developed for detection of zearalenone(ZEN) in cereals,in which ZEN conjugated to bovine serum albumin(BSA) is used as solid-phase antigen.A competitiv...A competitive indirect time-resolved fluoroimmunoassay(TRFIA) was developed for detection of zearalenone(ZEN) in cereals,in which ZEN conjugated to bovine serum albumin(BSA) is used as solid-phase antigen.A competitive indirect TRFIA was conducted by simultaneously incubating ZEN in standard or extracted samples with anti-ZEN monoclonal antibody over ZEN-BSA coated plates,and then determining the bound ZEN monoclonal antibody with goat anti-mouse europium conjugate.Samples were extracted with methanol/water...展开更多
Eu-chelate were used to construct a two-site sandwich-type assay for pepsinogen Ⅰ （PGI） with time-resolved fluoroimmunoassay （TRFIA） as a detection technique. On the noncompetitive assay, captured monoclonal anti...Eu-chelate were used to construct a two-site sandwich-type assay for pepsinogen Ⅰ （PGI） with time-resolved fluoroimmunoassay （TRFIA） as a detection technique. On the noncompetitive assay, captured monoclonal antibodies （McAbs） coated on wells were directed against a specific antigenic site on the PGI. Another McAbs, called as labeling McAbs, were prepared with the Eu-chelate of N-（p-isothiocyanatobenzyl）-diethylenetriamine-N, N, N, N-tetraacetic acid and directed against a different antigenic site on the PGI. The fluorescence counts of bound Eu^3＋ -McAbs were measured with the auto DELFIA1235 system. The PGI in sera from healthy volunteers were determined by PGI-TRFIA. The within-run and between-run CVs of the PGI-TRFIA were 1.9% and 4.7%, respectively, and the recovery rate was 102.65%. The assay had a detection limit of 0.05 μg· L^-1. The PGI-TRFIA provided a linear response from 3.5 to 328 μg· L^-1. The cross-reacting rate with pepsinogen Ⅱ was negligible. The linear correlation of PGI-TRFIA and radioimmunassay measurements resulted in a correlation coefficient of 0.977. The means of healthy volunteers were 154 ±43 μg·L^-1 for serum PGI. The availability of a highly sensitive, reliable, and convenient method for quantifying PGI will allow investigations into the possible diagnostic value of this analyte in various clinical conditions, including gastric carcinoma, duodenal ulcer, gastritis and severe atrophic gastritis.展开更多
A novel dual-label time-resolved fluoroimmunoassay method was developed for the simultaneous determination of chloramphenicol (CAP) and ractopamine (RAC) residues in 18 swine tissue samples,using anti-CAP and anti-RAC...A novel dual-label time-resolved fluoroimmunoassay method was developed for the simultaneous determination of chloramphenicol (CAP) and ractopamine (RAC) residues in 18 swine tissue samples,using anti-CAP and anti-RAC monoclonal antibodies labeled with europium (Eu 3+) and samarium (Sm 3+),respectively.The detection limits for CAP and RAC were 0.06 and 0.25ng/g.The recovery from swine muscle samples was 102%-121% for CAP at spiking levels of 0.1-5ng/g,and 69.8%-85.8% for RAC at spiking levels of 1-10ng/g.The results obtained from the swine tissue samples using this method showed good agreement with those obtained using ELISA and GC-MS,with correlation coefficients (R) between 0.92-0.98.展开更多
To quantitatively determine tylosin and tilmicosin in edible animal tissues,a time-resolved fluoroimmunoassay(TRFIA) has been developed and validated.For this purpose,desmycosin-O-carboxymethoxylamine-BSA was fixed on...To quantitatively determine tylosin and tilmicosin in edible animal tissues,a time-resolved fluoroimmunoassay(TRFIA) has been developed and validated.For this purpose,desmycosin-O-carboxymethoxylamine-BSA was fixed onto microtiter plates,standards and samples were loaded and,finally,diluted europium-labeled anti-tylosin antibodies were added.Results show that the limit of detection for tylosin was 0.03 ng mL-1 and that for tilmicosin was 0.05 ng mL-1.The recoveries were 73.6% to 120.5%,with coefficients of variation below 15.6% in various biological matrices spiked with tylosin and tilmicosin at concentrations of 50-200 ngg-1.There was good correlation(R2>0.99) between the TRFIA,an enzyme-linked immunosorbent assay and high performance liquid chromatography data.In conclusion,the new TRFIA is applicable to the detection of tylosin and tilmicosin and is an effective and economical method that will enable high-throughput sample screening.The method is expected to be widely applicable.展开更多
基金supported by the Innovation Fund for Technology Based Firms (06C26213201075)National High Technology Research and Development Program of China (863 Program) (2008AA102415)
文摘A competitive indirect time-resolved fluoroimmunoassay(TRFIA) was developed for detection of zearalenone(ZEN) in cereals,in which ZEN conjugated to bovine serum albumin(BSA) is used as solid-phase antigen.A competitive indirect TRFIA was conducted by simultaneously incubating ZEN in standard or extracted samples with anti-ZEN monoclonal antibody over ZEN-BSA coated plates,and then determining the bound ZEN monoclonal antibody with goat anti-mouse europium conjugate.Samples were extracted with methanol/water...
文摘Eu-chelate were used to construct a two-site sandwich-type assay for pepsinogen Ⅰ （PGI） with time-resolved fluoroimmunoassay （TRFIA） as a detection technique. On the noncompetitive assay, captured monoclonal antibodies （McAbs） coated on wells were directed against a specific antigenic site on the PGI. Another McAbs, called as labeling McAbs, were prepared with the Eu-chelate of N-（p-isothiocyanatobenzyl）-diethylenetriamine-N, N, N, N-tetraacetic acid and directed against a different antigenic site on the PGI. The fluorescence counts of bound Eu^3＋ -McAbs were measured with the auto DELFIA1235 system. The PGI in sera from healthy volunteers were determined by PGI-TRFIA. The within-run and between-run CVs of the PGI-TRFIA were 1.9% and 4.7%, respectively, and the recovery rate was 102.65%. The assay had a detection limit of 0.05 μg· L^-1. The PGI-TRFIA provided a linear response from 3.5 to 328 μg· L^-1. The cross-reacting rate with pepsinogen Ⅱ was negligible. The linear correlation of PGI-TRFIA and radioimmunassay measurements resulted in a correlation coefficient of 0.977. The means of healthy volunteers were 154 ±43 μg·L^-1 for serum PGI. The availability of a highly sensitive, reliable, and convenient method for quantifying PGI will allow investigations into the possible diagnostic value of this analyte in various clinical conditions, including gastric carcinoma, duodenal ulcer, gastritis and severe atrophic gastritis.
基金supported by the National Natural Science Foundation of China (21025729 and 20921063)
文摘A novel dual-label time-resolved fluoroimmunoassay method was developed for the simultaneous determination of chloramphenicol (CAP) and ractopamine (RAC) residues in 18 swine tissue samples,using anti-CAP and anti-RAC monoclonal antibodies labeled with europium (Eu 3+) and samarium (Sm 3+),respectively.The detection limits for CAP and RAC were 0.06 and 0.25ng/g.The recovery from swine muscle samples was 102%-121% for CAP at spiking levels of 0.1-5ng/g,and 69.8%-85.8% for RAC at spiking levels of 1-10ng/g.The results obtained from the swine tissue samples using this method showed good agreement with those obtained using ELISA and GC-MS,with correlation coefficients (R) between 0.92-0.98.
基金supported by the Scientific and Technological Research Project of Chongqing (CSTC2011ggB10009,cstc2012pt-kyys10002,cstc2012cx-rkxB10002)the National Natural Science Foundation of China(81202438)the Natural Science Foundation of the Chongqing Education Committee (KJ110605)
文摘To quantitatively determine tylosin and tilmicosin in edible animal tissues,a time-resolved fluoroimmunoassay(TRFIA) has been developed and validated.For this purpose,desmycosin-O-carboxymethoxylamine-BSA was fixed onto microtiter plates,standards and samples were loaded and,finally,diluted europium-labeled anti-tylosin antibodies were added.Results show that the limit of detection for tylosin was 0.03 ng mL-1 and that for tilmicosin was 0.05 ng mL-1.The recoveries were 73.6% to 120.5%,with coefficients of variation below 15.6% in various biological matrices spiked with tylosin and tilmicosin at concentrations of 50-200 ngg-1.There was good correlation(R2>0.99) between the TRFIA,an enzyme-linked immunosorbent assay and high performance liquid chromatography data.In conclusion,the new TRFIA is applicable to the detection of tylosin and tilmicosin and is an effective and economical method that will enable high-throughput sample screening.The method is expected to be widely applicable.
