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Objective To establish a fast and quantitative light initiated chemiluminescent assay (LICA) method for high mobility group box1 (HMGB1) determination.Methods Two strains of paired HMGB 1 monoclonal antibodies were used.One was used to coat receptor microspheres.The other was labeled with biotin first and then composed with chain mildew element of affinity donor microsphere to form LICA method for HMGB1.After optimizing the reaction system,the technical specifications of the method was evaluated.Serum HMGB1 levels of common pneumonia patients (CPP) and severe pneumonia patients (SPP) were measured and compared with that of health controls.Two-sample t test was used.Results The sensitivity of LICA was 0.1 μg/L,with linear measurement ranging from 0.1 to 1 000 μg/L.The precisions of intra-and inter-analysis were 1.74％-2.92％ and 1.93％-3.73％ respectively,both were lower than 5％.The recovery rate was 99.74％ (range:94.53％-106.37％).The correlation coefficient of LICA and enzyme-linked immunosorbent assay (ELISA) was 0.888 2.The LICA method had good specificity and no obvious cross reaction with HMGB2 and HMGB3.The serum HMGB1 level in CPP (n=35) and SPP (n=25) was significantly higher than that in health controls (n=35):(6.76±3.13),(19.69±+9.04) vs (1.49±+0.74) μg/L;t values:-5.447 and-5.186,both P＜0.01.The HMGB1 levels between CPP and SPP were also significantly different (t=-3.500,P＜0.01).Conclusions The established LICA method of HMGB1 has high sensitivity and specificity with reliable results.This method is also homogeneous,fast and cleaning-free,thus has a good prospect in clinical application.
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Objective To develop a time-resolved fluorescent microspheres immunochromatographic assay (TRFMIA) for detection of alphafetoprotein (AFP) and carcinoembryonic antigen (CEA) in human serum and to evaluate its performance.Methods The Eu-time-resolved fluorescent polystyrene particles conjugated with monoclonal antibody AC18# for AFP and AE03# for CEA were used as fluorescent labels.The monoclonal antibody AC17# for AFP,AE05# for CEA and goat anti-rabbit antibody were immobilized on the nitrocellulose membrane as the test lines and control line.Several performances indicators were measured,including linear range,detection limit,and specificity.AFP and CEA were measured by the new method and the results were compared with those obtained by time-resolved fluoroimmunoassay (TRFIA) and electro-chemiluminescence immunoassay (ECLIA) using linear correlation analysis.Results The measurement ranges of AFP were 0.07-1 000.00 kU/L with the intra-and inter-assay CV of 5.93％ and 11.07％,and those of CEA were 0.12-500.O0 μg/L with the intra-and inter-assay CV of 7.53％ and 12.13％ respectively.The average recovery rate of AFP and CEA was 92.77％ and 94.73％,respectively.Measurements obtained by TRFMIA had strong correlation coefficients ranging from 0.93 to 0.97 when compared with those obtained by TRFIA and ECLIA.Conclusion TRFMIA,which can simultaneously detect AFP and CEA,has been successfully established.
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Objective To establish a novel TRFIA for the measurement of heparanase (HPA) in serum samples,and investigate its clinical application.Methods The micro-pore plate wells were first coated with partially recombinant murine anti-human HPA monoclonal antibody.Biotin-labeled recombinant HPA protein was then used to compete with HPA in serum samples,and the prepared europium (III)-labeled streptavidin (Eu3+-SA) was used as signal readout for establishing the BSA-TRFIA assay.Using this assay,the serum HPA levels in healthy subjects (n=32) and tumor patients (n=54) were measured.The results of BSA-TRFIA were compared with those of ELISA.Two-sample t test (or t' test),and linear correlation analysis were used to analyze the data.Results The sensitivity of BSA-TRFIA for measuring HPA was 0.33 ug/L.The CV values for intra-batch and inter-batch were 5.29% and 7.54%,respectively.The average recovery rate was 105.5%.The standard curve range was 0-1 000 ug/L.The serum HPA level measured by the BSA-TRFIA method in healthy subjects was (2.03_+ 1.47) Iug/L.In tumor patients,the HPA level was significantly higher:(22.13_+7.38) ug/L (t'=19.388,P
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Objective To establish a method of TRFIA with high sensitivity and broad detecting range for serum HBV large surface protein (HBV-LP).Methods The monoclonal antibody of HBV-LP was covered on the microwell plate and incubated with HBV-LP in blood sample,then Eu3+ labeled antibody of HBs was added.HBV-LP in standard substance,blood samples of 66 chronic hepatitis B patients and 30 healthy controls was detected by the TRFIA and ELISA.x2 test and linear correlation analysis were used for data analysis.Results The dose-response curve of standard substance with TRFIA had good linear correlation (r=0.999).Normal reference range was established at 0-1.36 mg/L based on the ELISA results of 30 healthy controls.The sensitivity was 0.10 mg/L.The specificity was 100％ (30/30).Correlation coefficient between the TRFIA and the ELISA was 0.800 9 (P＜0.001).The positive detecting rates of the 2 methods were significantly different (89.4％(59/66) vs 77.3％(51/66),x2 =6.13,P＜0.01).The recovery rate for HBV-LP was between 95.93％-107.62％.The effective detecting range(CV＜10％) of TRFIA was 1.35-2 764.00 mg/L,and that of ELISA was 10.8-691.0 mg/L.Conclusion The TRFIA was established for HBV-LP detection with higher sensitivity and wider detecting range compared to ELISA.It has potential value for HBV screening and monitoring of antiviral therapy.
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Objective Statistically analyze the serum P53 antibody level in lung cancer patients to show its diagnostic values.Methods 53 cases of lung cancer and 32 cases of lung cancer after operation and chemotherapy are used as study group. 34 normal controls and 35 cases of lung benign disease were used as control group. All the serum P53 antibody levels of the testees are measured with a ELISA kit.Results In 53 cases of lung cancer group, the positive rate was 58.5% (31/53), while the average serum P53 antibody level was (4.551?6.074)IU/ml. In 32 cases of lung cancer after treatment including surgical and medical procedures, the positive rate was 25% (8/32), while the average serum P53 antibody level was (1.086?2.836)IU/ml. In 35 cases lung benign disease group, the average serum P53 antibody level was (0.163?0.097)IU/ml. In normal group, the average serum P53 antibody level was (0.162?0.08)IU/mL.The lung cancer group had statistical significance in comparison with the other three groups (P0.05).Conclusion The P53 antibody′s appearance is highly correlated with lung cancer. It can be used as an important marker of lung cancer and be beneficial to the differential diagnosis of lung lesions and the evaluation of the treatment.
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Objective To observe the variety of liver and kidney function during lung transplantation and discuss its reference value of diagnosis and therapy in clinical acute rejection(AR).Methods The variety of TBIL, ALT and CRE were dynamically studied and analyzed before and after lung transplantation in 2 cases shared one same donor's lung block.Results During the use of CSA,TBIL diversely procedurally raised and it got right when AR was under control; when illness was worsen,ALT abnormally raised and CRE had no remarkably change.Conclusion TBIL is the sensitivity index of hepatotoxicity of immunity inhibitor CSA after lung transplantation. The abnormal change of ALT is the index of disease turnover.