本文選題：HCV抗體　切入點(diǎn)：ELISA法　出處：《石河子大學(xué)》2015年碩士論文

【摘要】：目的:研究ELISA法檢測(cè)丙型肝炎病毒(HCV)抗體診斷陽(yáng)性的最佳Cut-off值及比較國(guó)內(nèi)常用四種試劑的敏感性和特異性。方法:1.高、低濃度的質(zhì)控品,平行檢測(cè)20次,計(jì)算批內(nèi)精密度變異系數(shù)(CV),并高、低濃度的質(zhì)控品,每天檢測(cè)復(fù)孔,連續(xù)檢測(cè)20天,計(jì)算批間精密度CV。2.選取2013年11月至2014年4月來(lái)我院住院患者或體檢者血清或血漿經(jīng)ELISA法初篩HCV抗體陽(yáng)性的標(biāo)本130例和接近臨界值(陰性)標(biāo)本45例(OD=0.15-2)。利用重組免疫印跡法(RIBA/PCR)確認(rèn)所有標(biāo)本的陰性或陽(yáng)性。3.判斷在試劑盒提供的Cut-off值的條件下,ELISA法與RIBA法檢測(cè)結(jié)果的一致性的Kappa值及ELISA法檢測(cè)HCV抗體的敏感性和特異性。4.利用SPSS17.0統(tǒng)計(jì)軟件繪制ROC曲線,選取最大的切點(diǎn)為臨界點(diǎn),并計(jì)算新的Cut-off值時(shí),ELISA法檢測(cè)結(jié)果與RIBA法結(jié)果的一致性Kappa值及ELISA法檢測(cè)HCV抗體的敏感性和特異性。5.選取實(shí)驗(yàn)室常用的四種試劑無(wú)序的編為A、B、C、D,在四種試劑檢測(cè)前評(píng)估評(píng)價(jià)批內(nèi)精密度CV和批間精密度CV,滿足實(shí)驗(yàn)室要求后,四種試劑分別檢測(cè)經(jīng)RIBA法確認(rèn)后的參考血清,計(jì)算A、B、C、D試劑的敏感性和特異性,并比較四種試劑檢測(cè)敏感性和特異性有無(wú)差異。結(jié)果:1.高、低濃度的質(zhì)控品批內(nèi)精密度分別為9.58%和10.08%,批間精密度分別為12.08%和13.78%。2.兩種檢測(cè)結(jié)果的一致性Kappa值為0.676,ELISA法檢測(cè)的敏感性為98.17%,特異性為65.16%,此時(shí),兩種檢測(cè)方法有統(tǒng)計(jì)學(xué)差異(P0.05)。3.利用ROC曲線確定的CO的臨界值Cut-off值為0.252,此時(shí),兩種檢測(cè)方法結(jié)果的一致性Kappa值為0.829。ELISA法檢測(cè)的敏感性為94.50%,特異性為89.39%,此時(shí),兩種檢測(cè)方法無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。4.利用ROC曲線確定S/CO的臨界值是2.44,此時(shí)的敏感性為93.6%,特異性為96.1%。5.四種試劑檢測(cè)前,評(píng)估的批內(nèi)精密度CV為9.82%,批間精密度CV為13.17%。6.A試劑的敏感性為99.08%,特異性為72.73%。B試劑的敏感性為99.08%,特異性為75.76%。C試劑的敏感性為99.08%,特異性為72.73%。D試劑的敏感性為99.08%,特異性為74.24%。且四種試劑間的敏感性、特異性無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論:1.試劑說(shuō)明書提供的Cut-off值可能與實(shí)驗(yàn)室實(shí)際Cut-off值存在差異。2.通過(guò)ROC曲線確定合適的Cut-off值,能有效的降低標(biāo)本檢測(cè)的假陽(yáng)性率,能夠?yàn)榕R床提供可靠的結(jié)果。3.國(guó)產(chǎn)試劑通常有較高的敏感性,不同的試劑間檢測(cè)的敏感性和特異性不同。4.建議不同的實(shí)驗(yàn)室應(yīng)根據(jù)所用的試劑,建立適合的Cut-off值,以減少標(biāo)本的漏診和誤診。
[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.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446.6

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本文編號(hào)：1686360