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Objective To use cyclic amplified fluorescence immunoassay method and electrochemiluminescence immunoassay method for quantitative detection of procalcitonin (PCT),and to evaluate the consistency of the test results.Methods With the method of electrochemiluminescence used as the contrast method and the cyclic enhanced fluorescence immunoassay method used as experimental method,samples of 219 hospitalized patients were measured in two ways.The results were divided into three groups:the serum group (Core serum group) and the micro blood group (Core micro blood group) which were detected in cyclic amplified fluorescence immunoassay method and the serum group (Roche serum group) which was detected by electrochemiluminescence immunoassay method.The data of three groups were compared with each other in paired t test,and correlation analysis,the relative sensitivity,the relative specificity,and Jorden index at the medical decision level (0.5 ng/mL and 2.0 ng/mL) were calculated and the Kappa Consistency Test was calculated.Results There were no statistical differences in three groups (P＞0.05).The Core serum group was positively correlated with the Roche serum group (r =0.993,P＜0.01).The linear regression equation was Y =-0.061+1.041X(0.04≤X≤60).The Core micro blood group was positively correlated with the Roche serum group(r=0.989,P＜0.01).The linear regression equation was Y=0.022+1.023X(0.04≤X≤60).The Core micro blood group was positively correlated with the Core serum group (r=0.986,P＜0.01).The linear regression equation was Y=0.129+0.973X(0.04≤X≤60).The relative sensitivity of the three comparison groups was greater than 92％ and the relative specificity was greater than 95％ at the medical decision level (0.5 ng/mL and 2.0 ng/mL),and the Jorden index and Kappa values were greater than 0.9.It indicated a better consistency.Conclusion Cyclic amplified fluorescence immunoassay and electrochemiluminescence immunoassay detection results were very consistent,which meet the clinical testing requirements.
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Immunogenic antigen (spinosin-BSA) and coating antigen (spinosin-OVA) of spinosin were synthesized by sodium periodate oxidation method. UV scanning analysis method showed that these two spinosins were successfully conjugated with carrier protein and the coupling ratio was 17 and 13.7, respectively. Meanwhile, when immunized by spinosin-BSA，the mice can produce anti-spinosin antibodies with the high titer (1:32 000),specificity (IC₅₀ 211.6 μg·L⁻¹) and low cross-reaction rate measured by ELISA tests. The artificial antigen of spinosin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
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Objective To investigate the comparation of sodium valproate concentration in peripheral blood monitoring by fluorescence polarization immunoassay method(FPIA)and high performance liquid chromatography method(HPLC)in epilepsy children.Methods 87 cases of epilepsy children received Sodium valproate treatment in our hospital from February 2014 to June 2016 were selected,fasting venous blood of elbow vein were collected the next morning after last medication, blood concentrations of Sodium valproate in serum samples were detected by FPIA method and HPLC method respectively,the correlation and consistency of results of the two methods were observed and compared.Results The intra day and inter day RSD of Sodium valproate concentration in peripheral blood in epilepsy children detected by FPIA were <5%,the recovery rate was 90%-110%,the precision and accuracy were high;The intra day and inter day RSD of Sodium valproate concentration in peripheral blood in epilepsy children detected by HPLC were <5%,the recovery rate was 90%-110%,the precision and accuracy were high; the linear regression equation between determination value of HPLC method (X) and determination value of FPIA method (Y) was:Y=0.8355X+1.8231,correlation coefficient r=0.914,the detection results were positively related;the Sodium valproate blood concentration detected by FPIA was significantly lower than that detected by HPLC method, the difference was statistically significant (P<0.05);Bland-Altman deviation chart results show that determination of blood drug concentration by HPLC method was higher than that of FPIA method by 7.2μg/mL.Conclusion The precision and accuracy of sodium valproate concentration in peripheral blood monitoring by FPIA method and HPLC method were all high, and the correlation was good, but the detection results of the two methods were significantly different,the detection result of HPLC method was higher than that of FPIA method,need to choose and judge according to the clinical situation.
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Objective: To synthesize and identify the artificial antigen of loganin for the first time, and provide a foundation for the preparation of specific monoclonal antibody and establishment of immunoassay method. Methods: Sodium periodate oxidation method was used to synthesize immunogenic antigen (loganin-BSA) and coating antigen (loganin-OVA) of loganin. Whether loganin was conjugated with BSA and OVA or not was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunised mice were detected by enzyme-linked immunosorbent assay (ELISA). Results: The results of MALDI-TOF-MS indicated that loganin was conjugated to BSA. The antibody against loganin obtained from immunised-mice could bind to loganin and the titer was up to 1:40 000. Conclusion: The artificial antigen of loganin was synthesized, which can be used further in the preparation of monoclonal antibody, application in quality control of Chinese materia medica and the pharmacokinetic study of loganin in laboratory animals.