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Objective To evaluate the clinical application effect of a specific antibody detection kit for human echinococcosis (hydatidosis kit),and provide technical support for further optimization of the production process of hydatidosis kit.Methods Using the method of retrospective investigation,1 481 patients with hydatidosis and 1 055 no-hydatidosis patients who were diagnosed by operation,pathology (gold standard) and hydatidosis kit in First Affiliated Hospital of Xinjiang Medical University,from 2012 to 2016 were selected.The clinical data was analyzed.The diagnostic performance of the kit was evaluated.The stepwise discriminant analysis method was applied to construct discriminant analysis function and establish a diagnostic model for echinococcosis.The detection efficiency of four antigens in hydatidosis kit was analyzed.Results A total of 2 536 patients [1 275 males,1 261 females,aged (41.62 ± 18.43) years old] were investigated,with the highest proportion in the 30-59 age group (1 489cases).Liver was the main organ affected by echinococcosis.The sensitivity,specificity and consistency of the hydatidosis kit were 94.80％ (1 404/1 481),71.00％ (749/1 055),and 84.90％ (2 153/2 536),Yoden index was 0.66 and Kappa value was 0.68.The stepwise discriminant analysis function Y =0.777X1 + 0.258X2 + 0.241X3-1.575 was constructed by the stepwise discriminant analysis method.There was no significant difference between the consistency of stepwise discriminant analysis model and the current diagnostic criteria (85.73％ vs 84.90％,x2 =0.694,P ＞ 0.05).The consistency of differential diagnosis between vesicular and cystic echinococcosis was 76.07％ (1 068/1 404).There was no significant difference in the detection efficacy between Echinococcus granulosus cystic fluid antigen (EgCF) and hydatidosis kit (P ＞ 0.05).Conclusions The diagnosis and differential diagnosis efficiency of hydatidosis kit is high.It is suitable for clinical diagnosis and field epidemiological investigation of echinococcosis in hospitals at all levels.EgCF can be used as the antigen of echinococcosis monoantigen strip,and can be applied in the epidemiological investigation of echinococcosis.
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Objective:To screen the appropriate anti-human IgG (secondary antibody) for conjugating with colloidal gold by dot immunogold filtration assay,and put forward the method of using multiple evaluation indicators for secondary antibody comparing and analyzing. Methods:Three different secondary antibodies A,B and C from three different companies were screened. Firstly the titration was used to determine the optimal reaction conditions. Then three secondary antibodies were conjugated to the colloidal gold,tested with a dot immunogold filtration assay for hydatidosis specific antibody detection. The optimal conjugated secondary antibody were compared with the standard indirect enzyme-linked immunosorbent assay. Results:The optimal pH of three secondary antibodies were 8. 5,the binding capacity were 38. 4,24 and 19. 2μg /ml colloidal gold. According to the comprehensive evaluation,the diagnostic effects of sec-ondary antibody B was better than others. Its diagnostic effects in dot immunogold filtration assay was compared with enzyme-linked im-munosorbent assay. The Kappa was 0. 895(P<0. 05) and showed the two methods were in good agreement. Conclusion:The appropriate secondary antibody for conjugating with colloidal gold could be screened by dot immunogold filtration assay and the evaluation indicators which put forward by this study,the screening method would have an important reference value for all kinds of colloidal gold test kit to screen the suitable secondary antibody.
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Objective To investigate the prevalence rate of ovine hepatic cystic echinococcosis (HCE) in sheep in Quaker Wusu area of Bayinbuluke of Xinjiang by ultrasonography and provide evidence for the prevention and control of HCE in sheep.Methods The prevalence screening of HCE in sheep was conducted based on ultrasound images in this area in July 2014.The sheep were divided into different groups by dental age to calculate the age specific prevalence rate of HCE and analyzed the correlation between the dental age and the prevalence rate.Results The total prevalence rate of HCE in sheep in this area was 36.9％.The prevalence rates of none-calcified HCE and calcified HCE were 7.3％ and 29.6％,respectively.The prevalence rates of none-calcified HCE in different age groups were 1.2％ (1-2 years old),1.4％ (2-3 years old),14.0％ (3-4 years old),10.0％ (4-5 years old),15.6％(5-6 years old) and 4.2％(＞6 years old) respectively.The prevalence rate of calcified HCE in different age groups were 9.9％(1-2 years old),16.2％ (2-3 years old),31.6％(3-4 years old),47.8％(4-5 years old),42.2％(5-6 years old) and 41.7％(＞6 years old) respectively.The prevalence rate of HCE in 1-2 years old group was lower than those in other groups,the prevalence rate of HCE in age groups ＞3 years increased significantly.There was positive correlation between the prevalence rate of HCE and dental age (r=0.372,R2=0.107,F=44.176,P=0.000).Conclusion HCE is highly endemic in Quaker Wusu area.The prevalence rate of HCE is low in sheep with young age and high in sheep aged 3-4 years.It is necessary to conduct early prevention of HCE in sheep in this area.
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Objective To establish a method to prepare anti-sodium estrone sulfate monoclonal antibody ( ESS-Mab) . Methods Balb/c mice were immunized by ESS. Immune methods were screened. The blood serum potencies were measured by indirect ELISA and the best consistence of antigen and the first antibody were confirmed with method of titration. Cell fusion was carried by using PEG method and McAb hybridoma was screened with the indirect ELISA. Results The best immunization method of mice was subcutaneously multi-point injection in mouse back with the dose of 200/100 μg ESS antigen five times. The fusion rate was 90. 2%. Hybridoma positive rate of ELISA screening was 4. 4%. Finally two cell lines 2C8 and 8A7 with good specificity and sensitivity were obtained. Conclusion The best immunization way is selected and indirect ELISA is set up effectively and reliably for screening and presenting ESS McAb. the hybridoma technique is able to prepare monoclonal antibody of anti-ESS successfully.
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Objective To investigate the expression and significance of Smad4 in peripheral hepatocytes of lesions in mice with hepatic alveolar echinococcosis.Methods Eight mice in the test group were inoculated with alveolar echinococcosis and 8 mice in the control group were injected with normal saline.The expression of Smad4 protein in the hepatic tissue of the mice was detected by immunohistochemistry method,and the data were analyzed by chi-square test.The effect of alveolar echinocoeeosis on the expression of Smad4 protein was investigated.Results Smad4 was detected in cell nuclei and partly in the cytoplasm.Six months after the establishment of the mice model for alveolar echinococeosis,the expression of Smad4 in the hepatic tissue in the test group was significantly higher than in the control group(x2=19.869,P＜0.05).The number of Smad4 with positive expression in the hepatocytes in the test group was slightly higher than in the control group,and the expression intensity of Smad4 in the test group was greater than in the control group(x2=58.3 10,P＜0.05).Conclusions The increase of the expression of Smad4 protein in the periphery hepatocytes and tissues of hepatic alveolar echinococcosis lesions may play a role in hepatic cirrhosis and immune evasion.