A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluore...A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in mi- cro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H2O2)-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL·1. The detection limit of CEA was 0.61 ng mL·1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation.展开更多
A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELI...A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).展开更多
DNA methyltransferase 1(DNMT1)is a useful biomarker for lung cancer in early clinical diagnosis.A rapid magnetic chemiluminescence immunoassay(MCLIA)for DNMT1 in human serum has been developed.Horseradish peroxidase(H...DNA methyltransferase 1(DNMT1)is a useful biomarker for lung cancer in early clinical diagnosis.A rapid magnetic chemiluminescence immunoassay(MCLIA)for DNMT1 in human serum has been developed.Horseradish peroxidase(HRP)-second-Ab was used to labeled polyclonal antibodies of anti-DNMT1.DNMT1 in sample integrates with specific immunomagnetic beads and can constitute a supersandwiched immunoreaction.In magnetic field,nonspecific materials can be separated.After luminescent substrate luminol-H2O2-BIP was added,the relative light unit(RLU)of HRP was detected and was discovered to be directly proportional to the content of DNMT1 in sample.The correlative variables involved in the MCLIA value were optimized and the methodological evaluation was carried out.After optimization,in the range of0.5–128 ng/mL,the linear regression equation was y=0.5014 x+1.769(x was logCDNMT1,y was relative luminescence units(RLU)/RLU0),and the limit of detection was 0.01 ng/mL.The RSD of intra-and interassays were 15.8%–16.9%and 14.3%–18.1%,respectively.The recovery was from 70.0%to 106.2%.Furthermore,paralleled with purchasable enzyme-linked immunosorbent assay(ELISA)kits,MCLEIA had lower detection limit,wider linear range and shorter detection time.Therefore,the MCLEIA established in this study could be used for the sensitive detection of DNMT1 in serum sample.展开更多
This letter reports a chemiluminescene immunoassay method combined with immunomagnetic separation to rapidly detect Cyfra 21-1, in which bio-functionlized magnetic nanocomposites were used as mobile substrate for capt...This letter reports a chemiluminescene immunoassay method combined with immunomagnetic separation to rapidly detect Cyfra 21-1, in which bio-functionlized magnetic nanocomposites were used as mobile substrate for capturing and isolating the cyfra 21-1 proteins. After the captured Cyfra 21-1 further reacted with horseradish peroxidase-conjugated anti-Cyfra 21-1 antibody to form a sandwich immunocomplex, the chemiluminescence would be produced as a result of addition of the chemiluminescent substrate. A home-made optical biosensor was designed to detect the chemiluminescence instead of other large instruments. There is a good linear response between the chemiluminescence intensity and the concentration of Cyfra 21-1 in the range from 0.2 to 50 ng/mL. The whole detection process including incubation, washing and detection could be performed within 45 min. The proposed method offers a simple, noninvasive and reliable tool for detecting non-small cell lung cancer and has potential application for clinical testing.展开更多
A high sensitive chemiluminescent magnetic enzyme-linked immunoassay method was established for Escherichia coli O157∶H7 determination.The bacterium antibody was labeled by alkaline phosphatase（ALP） that catalyzed ...A high sensitive chemiluminescent magnetic enzyme-linked immunoassay method was established for Escherichia coli O157∶H7 determination.The bacterium antibody was labeled by alkaline phosphatase（ALP） that catalyzed the decomposing of substrate 3-（2-spiroadamantane）-4-methoxy-4（3-phosphoryloxy）phenyl-1,2-dioxetane（AMPPD） to give the light emission.The sensitivity of the method is 8.5×104 Cell/mL with a linear range of 1.0×105—5.0×107 Cell/mL.The intra- and inter-assay CVs are 14.8% and 20.0% in pork samples, respectively.The correlation coefficient of present and the standard plate counting method is 0.981.The experiments with the spiked samples show that this method has great potential to be applied to（detecting） the concentration of Escherichia coli O157∶H7 in a variety of samples.展开更多
基金support from the CAS Major Scientific Research and Equipment Development Projects (YZ-0632)the National Natural Science Founolation of China (Grant No. 20907060)+2 种基金the CAS Graduate Innovation Foundation (No. YXLW-3)the National key Technology R&D Program (Grant No. 2006BAF07B03-1-1)Shaanxi Province, China’s Scholarship Council (Grant No. 08JK482)
文摘A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in mi- cro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H2O2)-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL·1. The detection limit of CEA was 0.61 ng mL·1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation.
基金supported by the National Basic Research Program of China (973 Program,no. 2007CB714507)National Nature Science Foundation of China (no. 90813015)
文摘A chemiluminescence enzyme immunoassay based on magnetic microparticles (MmPs-CLEIA) was developed to evaluate serum a-fetoprotein (AFP) in parallel with traditional colorimetric enzyme-linked immunosorbent assay (ELISA).A systematic comparison between the MmPs-CLEIA and colorimetric ELISA concluded that the MPs-CLEIA exhibited fewer dosages of immunoreagents,less total assay time,and better linearity,recovery,precision,sensitivity and validity.AFP was detected in forty human serum samples by the proposed MPs-CLEIA and ELISA,and the results were compared with commercial electrochemiluminescence immunoassay (ECLIA) kit.The correlation coefficient between MPs-CLEIA and ELISA was obtained with R 2 0.6703;however,the correlation between MPs-CLEIA and ECLIA (R 2 0.9582) was obviously better than that between colorimetric ELISA and ECLIA (R 2 0.6866).
基金supported by the National Natural Science Foundation of China(Nos.81402721,81573203,21605131)Science and Technology Department of Henan Province(No.22170004)
文摘DNA methyltransferase 1(DNMT1)is a useful biomarker for lung cancer in early clinical diagnosis.A rapid magnetic chemiluminescence immunoassay(MCLIA)for DNMT1 in human serum has been developed.Horseradish peroxidase(HRP)-second-Ab was used to labeled polyclonal antibodies of anti-DNMT1.DNMT1 in sample integrates with specific immunomagnetic beads and can constitute a supersandwiched immunoreaction.In magnetic field,nonspecific materials can be separated.After luminescent substrate luminol-H2O2-BIP was added,the relative light unit(RLU)of HRP was detected and was discovered to be directly proportional to the content of DNMT1 in sample.The correlative variables involved in the MCLIA value were optimized and the methodological evaluation was carried out.After optimization,in the range of0.5–128 ng/mL,the linear regression equation was y=0.5014 x+1.769(x was logCDNMT1,y was relative luminescence units(RLU)/RLU0),and the limit of detection was 0.01 ng/mL.The RSD of intra-and interassays were 15.8%–16.9%and 14.3%–18.1%,respectively.The recovery was from 70.0%to 106.2%.Furthermore,paralleled with purchasable enzyme-linked immunosorbent assay(ELISA)kits,MCLEIA had lower detection limit,wider linear range and shorter detection time.Therefore,the MCLEIA established in this study could be used for the sensitive detection of DNMT1 in serum sample.
基金supported by the National Natural Science Foundation of China (61125105, 61101048, 61002037, 61027001)the Major National Scientific Research Plan (2011CB933202)the"Strategic Priority Research Program"of the Chinese Academy of Sciences (XDA06020101)
文摘This letter reports a chemiluminescene immunoassay method combined with immunomagnetic separation to rapidly detect Cyfra 21-1, in which bio-functionlized magnetic nanocomposites were used as mobile substrate for capturing and isolating the cyfra 21-1 proteins. After the captured Cyfra 21-1 further reacted with horseradish peroxidase-conjugated anti-Cyfra 21-1 antibody to form a sandwich immunocomplex, the chemiluminescence would be produced as a result of addition of the chemiluminescent substrate. A home-made optical biosensor was designed to detect the chemiluminescence instead of other large instruments. There is a good linear response between the chemiluminescence intensity and the concentration of Cyfra 21-1 in the range from 0.2 to 50 ng/mL. The whole detection process including incubation, washing and detection could be performed within 45 min. The proposed method offers a simple, noninvasive and reliable tool for detecting non-small cell lung cancer and has potential application for clinical testing.
文摘A high sensitive chemiluminescent magnetic enzyme-linked immunoassay method was established for Escherichia coli O157∶H7 determination.The bacterium antibody was labeled by alkaline phosphatase（ALP） that catalyzed the decomposing of substrate 3-（2-spiroadamantane）-4-methoxy-4（3-phosphoryloxy）phenyl-1,2-dioxetane（AMPPD） to give the light emission.The sensitivity of the method is 8.5×104 Cell/mL with a linear range of 1.0×105—5.0×107 Cell/mL.The intra- and inter-assay CVs are 14.8% and 20.0% in pork samples, respectively.The correlation coefficient of present and the standard plate counting method is 0.981.The experiments with the spiked samples show that this method has great potential to be applied to（detecting） the concentration of Escherichia coli O157∶H7 in a variety of samples.
