ABSTRACT
Objective To detect the serum levels of calpainin (S100A11) using nanomagicbeabs sorting-time resolved fluoroimmuno assay (NMBS-TRFIA) and evaluate its diagnostic value in pancreatic carcinoma.Methods 88 patients with pancreatic carcinoma,50 patients with acute pancreatitis,10 patients with pancreatic cyst and 20 healthy controls were selected as the study subjects.The human peripheral serum blood was sorted with S100A11 antibody coupled nanomagicbeabs,and the concentration of S100A11 was detected by TRFIA method.The area under the receiver operating characteristic (ROC) curve (AUC) was used to determine the cut-off level for diagnosis of pancreatic carcinoma,in order to evaluate the sensitivity and specificity for diagnosis of pancreatic carcinoma.Results S100A11 showed a linear relationship within the range of 6.08～ 500 ng/ml using NMBS-TRFIA method,intraassay CV≤6.35％,inter-assay CV≤7.12％,and the average recovery rate was 104.7％.The serum levels of S100A11 in patients with pancreatic carcinoma,patients with acute pancreatitis and patients with pancreatic cyst were 185.53 ± 161.19,106.06±113.83 and 68.99± 47.83 ng/ml respectively.Compared with the normal control group (37.98±25.14 ng/ml),the differences were statistically significant (t=-8.065,-3.375,-2.266,all P ＜0.01).The serum levels of S100A11 in patients with pancreatic carcinoma was significantly higher than those in patients with acute pancreatitis and patients with pancreatic cyst (all P＜0.05).According to the ROC curve,ROCAUC=0.985 (95％ CI:0.972 ～ 0.997),the best cut-off level for the diagnosis of pancreatic carcinoma was 89.5 ng/ml (sensitivity 81.8 ％,specificity 67.5 ％).Conclusion NMBS-TRFIA can enrich S100A11 in serum and improve the detection sensitivity of serum S100A11,and the method is simple and easy to be popularized.Serum S100A11 has a high sensitivity and specificity in the diagnosis of pancreatic carcinoma,and is a new serum marker for the diagnosis of early pancreatic carcinoma.
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Objective To develop a kit of time?resolved fluoroimmunoassay(TRFIA)for detection of Schistosoma japonicum protein SjP38,and evaluate its effectiveness. Methods The anti 9G7 SjP38 monoclonal antibody was used as the capture anti?body coated with 96?hole plate,and the Eu3+labeled 1A6 monoclonal antibody was used as the detection antibody to establish the TRFIA SjP38 kit. In addition,the accuracy,sensitivity,precision,stability and coincidence rate to pathogenic diagnosis of the kit were evaluated. Results This established kit possessed high accuracy,wide linear range from 2 to 1 250 ng/ml,high sensitivity with the minimum detectable concentration of 0.14 ng/ml,and good precision(the coefficient variation of the intra?and inter?assay were 3.6%to 4.6%and 5.1%to 6.7%,respectively). The stability tests showed that the reagents could be stable for six months at 4℃,7 d at 37℃. The positive and negative corresponding rates to the pathogen detection method were 95%and 100%respectively. Conclusion All the performance and detection indicators of the kit have reached the requirements of clinical test,but its clinical application still needs further validation.
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Objective:To assess the efficacy of time-resolved fluoroimmuno assay(TRFIA) for quantative determination of?-fetoprotein(AFP) in comparison with radio immuno assay(RIA) and enzme linked immunoabsorbent assay(ELISA). Methods:Thirty serum samples from randomly selected patients were collected to test AFP content with TRFIA,RIA and ELISA respectively.Results:The linear range of AFP was 1 to l 000 ng/ml for TRFIA,5～400ng/ml for RIA and 5～300ng/ml for ELISA.The intraassay coefficients of variation(CV) and interassay CV were 3.3% and 4.7% for TRFIA,5.3% and 8.9% for RIA,and 10.5% and 15.2% for ELISA,respectively.Correlation tests revealed that the results of RIA were more closely correlated with those of TRFIA than with those of ELISA.Conclusion:TRFIA is shown to be a better method for AFP quantitation than RIA and ELISA in terms of precision and linear range.