甲型肝炎病毒总抗体检测试剂的比较及初步应用
发布时间：2018-08-06 08:35
【摘要】：甲型病毒性肝炎(甲肝)在我国目前划分为乙类传染病,由甲型肝炎病毒(Hepatitis A Virus, HAV)引起。甲肝经粪-口途径和直接接触传播,容易引起爆发与流行。HAV (Antibody to HAV, Anti-HAV)总抗体是流行病学调查中常用的重要指标.其阳性表明有HAV感染史或免疫史为了加强实验室Anti-HAV总抗体检测的质量控制,增加研究结果的可信程度,为甲型病毒性肝炎(甲肝)的血清流行病学检测试剂的选择提供参考,本研究对市售的Anti-HAV总抗体检测试剂进行评价与比较并在血清流行病学上作初步应用。 通过拟合并选择最优的标准曲线,建立Anti-HAV总抗体的定量方法。将Anti-HAV总抗体标准品倍比稀释,用美国雅培(Abbott)公司生产的微粒子酶免疫测定法检测试剂盒HAVAb2.0依次检测五遍。以标准品的Anti-HAV总抗体浓度X为自变量,以测定值CO/S为应变量,采用常用对数转换和自然对数转换两种转换方式,组合线性模型、指数模型、logistic模型三种拟合模型进行标准曲线拟合,得到六个回归方程,均有统计学意义(F检验,P0.001),其中两种转换方式的logistic回归模型的R2最高,为0.9893。各个浓度标准品测定的均值代入两个logistic回归方程得到的误差平均值最小。三种标准曲线拟合模型中,logistic回归的拟合效果最好。并没有观察到自然对数和常用对数两种数据变换方式对拟合的效果的影响。 本研究建立了Anti-HAV,总抗体的参比系统,作为不同试剂比较的共同平台。对314份血清预备参比品,在采用“即刻法”进行统计学室内质量控制合格的情况下,用Abbott公司的HAVAb2.0试剂检测确定其阴阳性,并对检测试剂灰区的预备参比品重复检测三次,将定性结果不一致的预备参比品剔除。然后对纳入的311份预备参比品进行定量并根据定量结果对参比系统的内部构成进行调整。将其中过多的高浓度参比品去除一部分,再通过稀释参比品的方式增加弱阳性参比品的数量。最终建立的参比系统中共有71份阳性参比品,其中抗体浓度10OOOmIU/ml的12份,1000～10OOOmIU/ml的11份,100～1000mIU/ml的37份,100mIU/ml的11份；阴性64份。稀释的参比品抗体浓度为100～500mIU/ml的参比品17份,100mIU/ml的18份,7份阴性。 选两种国内常用的Anti-HAV总抗体检测的酶免疫检测试剂进行评价比较。用A、B两试剂对本研究中建立的参比系统平行检测各8遍,记录CO/S值并作ln(CO/S+0.1)转换,通过广义线性化估计方程(Generalized Estimating Equation,GEE)模型估计各次测试样本的阳性概率,计算比较两种试剂的可靠性指标和真实性指标。A试剂的组内相关系数(Intraclass Correlation Coefficient, ICC)和变异系数(Coefficient of Variability.CV)分别为0.9971和5.7840%,B试剂的为0.9952和6.2931%(Bootstrap法.P0.05)；A试剂的全部受试者工作特征(Receive Operation Characteristic, ROC)曲线下面积(Area Under Curve, AUC)和部分(Partial) ROC曲线(特异度为0.9～1)下面积(pAUC)分别为0.9557和0.0717,B的为0.9404和0.0663(Bootstrap法,P0.05)；在特异度为0.920～0.970时,间隔0.005计算的固定特异度下的灵敏度,在特异度为0.945、0.950、0.955、0.960几个观察点时,A试剂的灵敏度B试剂(Bootstrap法,P0.05)；但在特异度为其他观察点时,对应灵敏度的差异无统计学意义(Bootstrap法,P0.05)。将CO/S值转为二分类定性资料,得到A试剂的粗符合率(Crude Accuracy, CA)和Kappa值分别为92.66%和0.85,B试剂的为90.40%和0.80,A、B两试剂的灵敏度分别为94.34%和88.68%,特异度分别为90.14%和92.96%,Youden指数为0.84和0.81。差异均无统计学意义(Z检验,P0.05)。可以看出,A、B两种试剂均表现出良好的诊断能力,虽然某些指标统计学上有差异,但差异有无实际意义值得进一步探讨,在实际应用时,可根据具体情况进行选择。 在Anti-HAV总抗体检测试剂比较的基础上,我们对内蒙古巴彦淖尔市乌拉特前旗地区蒙古族和汉族中学生587人进行初步的血清学调查,得到总体阳性率为14.31%,对年龄、性别、民族这几个因素用logistic回归分析,得到不同年龄段Anti-HAV总抗体阳性率不同。提示应在该地青少年中加强甲肝疫苗的接种,提高Anti-HAV阳性率,防止甲肝的爆发或流行。
[Abstract]:Hepatitis A (hepatitis A) is divided into B infectious disease in China, which is caused by hepatitis A virus (Hepatitis A Virus, HAV). It is easy to cause the outbreak and epidemic of.HAV (Antibody to HAV, Anti-HAV) as an important index in epidemiological investigation. The positive indicates that there is HA. V infection history or immunization history is to strengthen the quality control of Anti-HAV total antibody test in laboratory, increase the credibility of the research results, and provide reference for the selection of sero epidemiological detection reagents for hepatitis A (hepatitis A). This study evaluates and compares the Anti-HAV total anti physical test agents sold in the market and in sero epidemiology. Preliminary application.
A quantitative method for the Anti-HAV total antibody was established by combining the optimal standard curve. The double ratio of the standard Anti-HAV antibody was diluted, and the reagent kit HAVAb2.0 was detected five times by the microparticle enzyme immunoassay method produced by American Abbott (Abbott) company. The Anti-HAV total antibody concentration X of the standard product was independent variable to determine the value CO/S. For the dependent variable, two kinds of conversion methods used in common logarithmic conversion and natural logarithm conversion, combined linear model, exponential model, and logistic model three fitting models to fit the standard curve, get six regression equations, all of which have statistical significance (F test, P0.001), of which the logistic regression model of two transformation modes is the highest R2, 0.9893. The mean value of the mean value of each concentration standard sample into two logistic regression equations is the least. In the three standard curve fitting models, the fitting effect of logistic regression is the best. No effect of the natural logarithms and the two common logarithm data transformation methods on the results of the fitting results is not observed.
In this study, the reference system of Anti-HAV and total antibody was established as a common platform for comparison of different reagents. In the case of 314 serum preparatory reference materials, under the condition that the quality control was qualified by "instant method", the HAVAb2.0 reagent of Abbott company was used to determine its Yin and Yang, and the preparatory reference of the test reagent grey area was obtained. Repeat the test three times, remove the preparatory reference material with inconsistent qualitative results, then adjust the internal composition of the reference system according to the quantitative results of the 311 prepared ginseng, and remove a part of the high concentration of the high concentration of the reference, and then increase the number of the weakly positive reference by the dilution reference. In the final system, there were 71 positive reference materials, including 12 10OOOmIU/ml, 11, 100 to 1000mIU/ml, 11, 64, 100 to 500mIU/ml, 17, 18 and 7 negative.
The evaluation and comparison of two kinds of enzyme immunoassay reagents for the detection of Anti-HAV total antibody were compared. The reference system established in this study was detected by A and B two, the CO/S value was recorded and the LN (CO/S+0.1) conversion was recorded, and the test samples were estimated by the generalized linearization estimation equation (Generalized Estimating Equation, GEE) model. The positive probability of the two reagents, the reliability index of the two reagents and the authenticity index, the intra group correlation coefficient (Intraclass Correlation Coefficient, ICC) and the coefficient of variation (Coefficient of Variability.CV) were 0.9971 and 5.7840% respectively, the B reagents were 0.9952 and 6.2931% (Bootstrap.P0.05), and all the A agents were tested. The area under the Receive Operation Characteristic (ROC) curve (Area Under Curve, AUC) and some (Partial) ROC curves (the specificity of 0.9 to 1) are 0.9557 and 0.0717 respectively, respectively, and B are 0.9404 and 0.0663, and the sensitivity under the fixed special degree of 0.005 in the interval 0.005 is 0.920 to 0.970. Degree, the sensitivity of A reagent B reagent (Bootstrap method, P0.05) when the specificity is 0.945,0.950,0.955,0.960, but when the specificity is other observation point, the difference of sensitivity is not statistically significant (Bootstrap, P0.05). CO/S value is converted to two classification qualitative data, and the coarse coincidence rate of A reagents (Crude Accuracy, CA) is obtained. Kappa values were 92.66% and 0.85, B reagents 90.40% and 0.80, A, B two reagents were 94.34% and 88.68% respectively, the specificity was 90.14% and 92.96%, the Youden index was 0.84 and 0.81. (Z test, P0.05). It can be seen that A, B two reagents all showed good diagnostic ability, although some index Statistics There are differences in learning, but whether the difference has practical significance is worth further exploring. In practical application, it can be selected according to specific circumstances.
On the basis of the comparison of the Anti-HAV total antibody test reagents, we conducted a preliminary serological survey on 587 Mongolian and Han Middle School Students in Bayannaoer, Inner Mongolia, and obtained a total positive rate of 14.31%. The factors of age, sex and nationality were analyzed by logistic regression, and the total resistance of Anti-HAV in different ages was obtained. It is suggested that the inoculation of hepatitis A vaccine should be strengthened among adolescents in this area to increase the positive rate of anti-HAV and prevent the outbreak or epidemic of hepatitis A.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.61
本文编号：2167152
[Abstract]:Hepatitis A (hepatitis A) is divided into B infectious disease in China, which is caused by hepatitis A virus (Hepatitis A Virus, HAV). It is easy to cause the outbreak and epidemic of.HAV (Antibody to HAV, Anti-HAV) as an important index in epidemiological investigation. The positive indicates that there is HA. V infection history or immunization history is to strengthen the quality control of Anti-HAV total antibody test in laboratory, increase the credibility of the research results, and provide reference for the selection of sero epidemiological detection reagents for hepatitis A (hepatitis A). This study evaluates and compares the Anti-HAV total anti physical test agents sold in the market and in sero epidemiology. Preliminary application.
A quantitative method for the Anti-HAV total antibody was established by combining the optimal standard curve. The double ratio of the standard Anti-HAV antibody was diluted, and the reagent kit HAVAb2.0 was detected five times by the microparticle enzyme immunoassay method produced by American Abbott (Abbott) company. The Anti-HAV total antibody concentration X of the standard product was independent variable to determine the value CO/S. For the dependent variable, two kinds of conversion methods used in common logarithmic conversion and natural logarithm conversion, combined linear model, exponential model, and logistic model three fitting models to fit the standard curve, get six regression equations, all of which have statistical significance (F test, P0.001), of which the logistic regression model of two transformation modes is the highest R2, 0.9893. The mean value of the mean value of each concentration standard sample into two logistic regression equations is the least. In the three standard curve fitting models, the fitting effect of logistic regression is the best. No effect of the natural logarithms and the two common logarithm data transformation methods on the results of the fitting results is not observed.
In this study, the reference system of Anti-HAV and total antibody was established as a common platform for comparison of different reagents. In the case of 314 serum preparatory reference materials, under the condition that the quality control was qualified by "instant method", the HAVAb2.0 reagent of Abbott company was used to determine its Yin and Yang, and the preparatory reference of the test reagent grey area was obtained. Repeat the test three times, remove the preparatory reference material with inconsistent qualitative results, then adjust the internal composition of the reference system according to the quantitative results of the 311 prepared ginseng, and remove a part of the high concentration of the high concentration of the reference, and then increase the number of the weakly positive reference by the dilution reference. In the final system, there were 71 positive reference materials, including 12 10OOOmIU/ml, 11, 100 to 1000mIU/ml, 11, 64, 100 to 500mIU/ml, 17, 18 and 7 negative.
The evaluation and comparison of two kinds of enzyme immunoassay reagents for the detection of Anti-HAV total antibody were compared. The reference system established in this study was detected by A and B two, the CO/S value was recorded and the LN (CO/S+0.1) conversion was recorded, and the test samples were estimated by the generalized linearization estimation equation (Generalized Estimating Equation, GEE) model. The positive probability of the two reagents, the reliability index of the two reagents and the authenticity index, the intra group correlation coefficient (Intraclass Correlation Coefficient, ICC) and the coefficient of variation (Coefficient of Variability.CV) were 0.9971 and 5.7840% respectively, the B reagents were 0.9952 and 6.2931% (Bootstrap.P0.05), and all the A agents were tested. The area under the Receive Operation Characteristic (ROC) curve (Area Under Curve, AUC) and some (Partial) ROC curves (the specificity of 0.9 to 1) are 0.9557 and 0.0717 respectively, respectively, and B are 0.9404 and 0.0663, and the sensitivity under the fixed special degree of 0.005 in the interval 0.005 is 0.920 to 0.970. Degree, the sensitivity of A reagent B reagent (Bootstrap method, P0.05) when the specificity is 0.945,0.950,0.955,0.960, but when the specificity is other observation point, the difference of sensitivity is not statistically significant (Bootstrap, P0.05). CO/S value is converted to two classification qualitative data, and the coarse coincidence rate of A reagents (Crude Accuracy, CA) is obtained. Kappa values were 92.66% and 0.85, B reagents 90.40% and 0.80, A, B two reagents were 94.34% and 88.68% respectively, the specificity was 90.14% and 92.96%, the Youden index was 0.84 and 0.81. (Z test, P0.05). It can be seen that A, B two reagents all showed good diagnostic ability, although some index Statistics There are differences in learning, but whether the difference has practical significance is worth further exploring. In practical application, it can be selected according to specific circumstances.
On the basis of the comparison of the Anti-HAV total antibody test reagents, we conducted a preliminary serological survey on 587 Mongolian and Han Middle School Students in Bayannaoer, Inner Mongolia, and obtained a total positive rate of 14.31%. The factors of age, sex and nationality were analyzed by logistic regression, and the total resistance of Anti-HAV in different ages was obtained. It is suggested that the inoculation of hepatitis A vaccine should be strengthened among adolescents in this area to increase the positive rate of anti-HAV and prevent the outbreak or epidemic of hepatitis A.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.61
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