甲型肝炎病毒總抗體檢測試劑的比較及初步應用
發(fā)布時間：2018-08-06 08:35
【摘要】：甲型病毒性肝炎(甲肝)在我國目前劃分為乙類傳染病,由甲型肝炎病毒(Hepatitis A Virus, HAV)引起。甲肝經糞-口途徑和直接接觸傳播,容易引起爆發(fā)與流行。HAV (Antibody to HAV, Anti-HAV)總抗體是流行病學調查中常用的重要指標.其陽性表明有HAV感染史或免疫史為了加強實驗室Anti-HAV總抗體檢測的質量控制,增加研究結果的可信程度,為甲型病毒性肝炎(甲肝)的血清流行病學檢測試劑的選擇提供參考,本研究對市售的Anti-HAV總抗體檢測試劑進行評價與比較并在血清流行病學上作初步應用。 通過擬合并選擇最優(yōu)的標準曲線,建立Anti-HAV總抗體的定量方法。將Anti-HAV總抗體標準品倍比稀釋,用美國雅培(Abbott)公司生產的微粒子酶免疫測定法檢測試劑盒HAVAb2.0依次檢測五遍。以標準品的Anti-HAV總抗體濃度X為自變量,以測定值CO/S為應變量,采用常用對數轉換和自然對數轉換兩種轉換方式,組合線性模型、指數模型、logistic模型三種擬合模型進行標準曲線擬合,得到六個回歸方程,均有統(tǒng)計學意義(F檢驗,P0.001),其中兩種轉換方式的logistic回歸模型的R2最高,為0.9893。各個濃度標準品測定的均值代入兩個logistic回歸方程得到的誤差平均值最小。三種標準曲線擬合模型中,logistic回歸的擬合效果最好。并沒有觀察到自然對數和常用對數兩種數據變換方式對擬合的效果的影響。 本研究建立了Anti-HAV,總抗體的參比系統(tǒng),作為不同試劑比較的共同平臺。對314份血清預備參比品,在采用“即刻法”進行統(tǒng)計學室內質量控制合格的情況下,用Abbott公司的HAVAb2.0試劑檢測確定其陰陽性,并對檢測試劑灰區(qū)的預備參比品重復檢測三次,將定性結果不一致的預備參比品剔除。然后對納入的311份預備參比品進行定量并根據定量結果對參比系統(tǒng)的內部構成進行調整。將其中過多的高濃度參比品去除一部分,再通過稀釋參比品的方式增加弱陽性參比品的數量。最終建立的參比系統(tǒng)中共有71份陽性參比品,其中抗體濃度10OOOmIU/ml的12份,1000～10OOOmIU/ml的11份,100～1000mIU/ml的37份,100mIU/ml的11份；陰性64份。稀釋的參比品抗體濃度為100～500mIU/ml的參比品17份,100mIU/ml的18份,7份陰性。 選兩種國內常用的Anti-HAV總抗體檢測的酶免疫檢測試劑進行評價比較。用A、B兩試劑對本研究中建立的參比系統(tǒng)平行檢測各8遍,記錄CO/S值并作ln(CO/S+0.1)轉換,通過廣義線性化估計方程(Generalized Estimating Equation,GEE)模型估計各次測試樣本的陽性概率,計算比較兩種試劑的可靠性指標和真實性指標。A試劑的組內相關系數(Intraclass Correlation Coefficient, ICC)和變異系數(Coefficient of Variability.CV)分別為0.9971和5.7840%,B試劑的為0.9952和6.2931%(Bootstrap法.P0.05)；A試劑的全部受試者工作特征(Receive Operation Characteristic, ROC)曲線下面積(Area Under Curve, AUC)和部分(Partial) ROC曲線(特異度為0.9～1)下面積(pAUC)分別為0.9557和0.0717,B的為0.9404和0.0663(Bootstrap法,P0.05)；在特異度為0.920～0.970時,間隔0.005計算的固定特異度下的靈敏度,在特異度為0.945、0.950、0.955、0.960幾個觀察點時,A試劑的靈敏度B試劑(Bootstrap法,P0.05)；但在特異度為其他觀察點時,對應靈敏度的差異無統(tǒng)計學意義(Bootstrap法,P0.05)。將CO/S值轉為二分類定性資料,得到A試劑的粗符合率(Crude Accuracy, CA)和Kappa值分別為92.66%和0.85,B試劑的為90.40%和0.80,A、B兩試劑的靈敏度分別為94.34%和88.68%,特異度分別為90.14%和92.96%,Youden指數為0.84和0.81。差異均無統(tǒng)計學意義(Z檢驗,P0.05)。可以看出,A、B兩種試劑均表現出良好的診斷能力,雖然某些指標統(tǒng)計學上有差異,但差異有無實際意義值得進一步探討,在實際應用時,可根據具體情況進行選擇。 在Anti-HAV總抗體檢測試劑比較的基礎上,我們對內蒙古巴彥淖爾市烏拉特前旗地區(qū)蒙古族和漢族中學生587人進行初步的血清學調查,得到總體陽性率為14.31%,對年齡、性別、民族這幾個因素用logistic回歸分析,得到不同年齡段Anti-HAV總抗體陽性率不同。提示應在該地青少年中加強甲肝疫苗的接種,提高Anti-HAV陽性率,防止甲肝的爆發(fā)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.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R512.61
本文編號：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.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R512.61
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