ELISA法检测HCV抗体最佳Cut-off的确定及四种试剂比较

发布时间：2018-03-30 14:41

本文选题：HCV抗体　切入点：ELISA法　出处：《石河子大学》2015年硕士论文

【摘要】：目的:研究ELISA法检测丙型肝炎病毒(HCV)抗体诊断阳性的最佳Cut-off值及比较国内常用四种试剂的敏感性和特异性。方法:1.高、低浓度的质控品,平行检测20次,计算批内精密度变异系数(CV),并高、低浓度的质控品,每天检测复孔,连续检测20天,计算批间精密度CV。2.选取2013年11月至2014年4月来我院住院患者或体检者血清或血浆经ELISA法初筛HCV抗体阳性的标本130例和接近临界值(阴性)标本45例(OD=0.15-2)。利用重组免疫印迹法(RIBA/PCR)确认所有标本的阴性或阳性。3.判断在试剂盒提供的Cut-off值的条件下,ELISA法与RIBA法检测结果的一致性的Kappa值及ELISA法检测HCV抗体的敏感性和特异性。4.利用SPSS17.0统计软件绘制ROC曲线,选取最大的切点为临界点,并计算新的Cut-off值时,ELISA法检测结果与RIBA法结果的一致性Kappa值及ELISA法检测HCV抗体的敏感性和特异性。5.选取实验室常用的四种试剂无序的编为A、B、C、D,在四种试剂检测前评估评价批内精密度CV和批间精密度CV,满足实验室要求后,四种试剂分别检测经RIBA法确认后的参考血清,计算A、B、C、D试剂的敏感性和特异性,并比较四种试剂检测敏感性和特异性有无差异。结果:1.高、低浓度的质控品批内精密度分别为9.58%和10.08%,批间精密度分别为12.08%和13.78%。2.两种检测结果的一致性Kappa值为0.676,ELISA法检测的敏感性为98.17%,特异性为65.16%,此时,两种检测方法有统计学差异(P0.05)。3.利用ROC曲线确定的CO的临界值Cut-off值为0.252,此时,两种检测方法结果的一致性Kappa值为0.829。ELISA法检测的敏感性为94.50%,特异性为89.39%,此时,两种检测方法无统计学差异(P0.05)。4.利用ROC曲线确定S/CO的临界值是2.44,此时的敏感性为93.6%,特异性为96.1%。5.四种试剂检测前,评估的批内精密度CV为9.82%,批间精密度CV为13.17%。6.A试剂的敏感性为99.08%,特异性为72.73%。B试剂的敏感性为99.08%,特异性为75.76%。C试剂的敏感性为99.08%,特异性为72.73%。D试剂的敏感性为99.08%,特异性为74.24%。且四种试剂间的敏感性、特异性无统计学差异(P0.05)。结论:1.试剂说明书提供的Cut-off值可能与实验室实际Cut-off值存在差异。2.通过ROC曲线确定合适的Cut-off值,能有效的降低标本检测的假阳性率,能够为临床提供可靠的结果。3.国产试剂通常有较高的敏感性,不同的试剂间检测的敏感性和特异性不同。4.建议不同的实验室应根据所用的试剂,建立适合的Cut-off值,以减少标本的漏诊和误诊。
[Abstract]:Objective: to study the best Cut-off value of ELISA method for the detection of hepatitis C virus (HCV) antibody and to compare the sensitivity and specificity of four kinds of reagents commonly used in China. The intra-batch precision coefficient of variation (CV) was calculated, and high and low concentration quality control products were detected daily and continuously for 20 days. From November 2013 to April 2014, 130 samples of serum or plasma positive for HCV antibody were screened by ELISA method, and 45 samples with close to critical value (negative) were selected. The recombinant immunoassay was used to detect HCV antibody in sera or plasma of patients in our hospital from November 2013 to April 2014. Epidemic blotting confirmed that all specimens were negative or positive .3.The Kappa value of Elisa and RIBA method was consistent with that of RIBA method, and the sensitivity and specificity of HCV antibody detected by ELISA method. 4. SPSS17.0 was used to determine the sensitivity and specificity of HCV antibody. The software plotted the ROC curve, Select the maximum tangent point as the critical point, At the same time, when calculating the new Cut-off value, the result of Elisa is consistent with the result of RIBA method, and the sensitivity and specificity of ELISA method to detect HCV antibody. 5. Select four kinds of reagents that are used in the laboratory to be randomly classified as Agna Bu Con D, and evaluate them before the detection of the four kinds of reagents. After evaluating the intra-and inter-batch precision CV and meeting the laboratory requirements, The four kinds of reagents were used to detect the reference serum confirmed by RIBA method, to calculate the sensitivity and specificity of the reagent, and to compare the sensitivity and specificity of the four reagents. The intra-assay precision of low concentration quality control products was 9.58% and 10.08%, and the inter-assay precision was 12.08% and 13.78.2. the Kappa value of the two results was 0.676. The sensitivity and specificity of Elisa were 98.1717 and 65.16g, respectively. The critical value of CO determined by ROC curve was 0.252, and the consistent Kappa value of the two methods was 94.50 and 89.39, respectively. At this time, the sensitivity and specificity of the two methods were 94.50 and 89.39, respectively. There was no statistical difference between the two methods. The critical value of S/CO determined by ROC curve was 2.44. The sensitivity and specificity of the two methods were 93.6and 96.1g 路5. before the four reagents were tested, The intra-assay precision CV was 9.82, the inter-assay precision CV was 13.17.6.A, the sensitivity was 99.08, the specificity was 72.73.B, the sensitivity was 99.08. the specificity was 75.76.C, the sensitivity was 99.08. the specificity was 72.73.D, the sensitivity was 99.08. 74.24 sensitivity between the four reagents, There was no statistical difference in specificity (P 0.05). Conclusion 1. The Cut-off value provided in the reagent specification may be different from the actual Cut-off value in laboratory. 2. Determining the appropriate Cut-off value by ROC curve can effectively reduce the false positive rate of specimen detection. Domestic reagents usually have high sensitivity, and different reagents have different sensitivity and specificity. 4. It is suggested that different laboratories should establish suitable Cut-off values according to the reagents used. In order to reduce the missed diagnosis and misdiagnosis.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6

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