【摘要】：目的建立人血清中抗A、C、Y、W135群腦膜炎球菌莢膜多糖Ig G抗體含量的ELISA定量檢測方法,并進行驗證。方法篩選適用的酶標板,優(yōu)化抗原包被濃度,確定ELISA檢測過程的關(guān)鍵條件,對建立的ELISA方法的特異性、線性、準確性、精密度、最低檢測值和穩(wěn)定性進行驗證,并建立實驗室質(zhì)控血清的質(zhì)控檢測范圍。結(jié)果酶標板的板內(nèi)CV為9.8%,板間CV為16.4%,符合檢測使用要求。免疫血清經(jīng)多糖吸收后,群特異性抗體含量與同群多糖吸收劑量呈明顯的劑量依賴關(guān)系。12份美國CDC質(zhì)控血清中各群Ig G抗體含量與血清的稀釋度呈負相關(guān),相關(guān)系數(shù)(r)和斜率(slope)均接近-1,線性關(guān)系良好;檢測結(jié)果與CDC公布數(shù)據(jù)一致性良好,一致性相關(guān)系數(shù)A、C、W135、Y群分別為0.912 5、0.939 6、0.963 7和0.920 6,準確度較好。實驗室內(nèi)部質(zhì)控血清Men20中各群Ig G抗體含量試驗內(nèi)和試驗間精密度分別小于10%和20%,精密度較好;各血清群別的CV值均30%,穩(wěn)定性較好;A、C、Y、W135群多糖Ig G抗體含量最低檢測值分別為0.092、0.118、0.067和0.035μg/ml,質(zhì)控血清的檢測范圍分別為:92.49~36.53、37.29~17.69、66.18~25.54和50.15~23.31μg/ml。結(jié)論建立了標準化的檢測人血清中抗A、C、Y、W135群腦膜炎球菌多糖Ig G抗體含量的定量ELISA方法,并確定了實驗室日常用質(zhì)控血清Men20的質(zhì)控檢測值范圍。
[Abstract]:Aim to establish a ELISA method for quantitative determination of anti A, C, Y, W135 meningococcal capsule polysaccharide Ig G antibody in human serum and verify it. Methods the suitable enzyme plate was screened, the concentration of antigen coated was optimized, the key conditions of ELISA detection process were determined, and the specificity, linearity, accuracy, precision, minimum detection value and stability of the established ELISA method were verified. And establish the laboratory quality control serum quality control detection range. Results the intraplate CV and inter-plate CV were 9.8% and 16.4% respectively. There was a significant dose-dependent relationship between the group-specific antibody content and the absorbed dose of the same-group polysaccharide. There was a negative correlation between the content of each group of Ig-G antibody and the dilution of the serum in the 12 CDC quality-controlled sera of the United States. The correlation coefficient (r) and slope (slope) are both close to-1, and the linear relationship is good. The correlation coefficients A, C, W135 and Y were 0.912, 0.939 6, 0.963 7 and 0.920 6, respectively. The results were in good agreement with CDC published data. The intra-and inter-test precision of each group of Ig-G antibody in the internal quality control serum Men20 was less than 10% and 20%, respectively, and the precision was better, the CV value of each serum group was 30%, and the stability was good. The lowest detection values of Ig G antibody in A, C, Y, W135 polysaccharide groups were 0.092, 0.118, 0.067 and 0.035 渭 g / ml, respectively, the detection ranges were 92.49 渭 g / ml, 37.29 渭 g / ml., 66.18 渭 g / ml. and 50.15 ~ 23.31 渭 g / ml., respectively. Conclusion A standardized ELISA method was established for the determination of anti-A, C, Y, W135 group meningococcal polysaccharide Ig G antibody in human serum, and the range of quality control detection value of Men20 in laboratory routine quality control serum was determined. [WT5 "HZ] conclusion\ [WT5" BZ]
【作者單位】： 蘭州生物制品研究所有限責任公司甘肅省疫苗工程技術(shù)研究中心;中國食品藥品檢定研究院;美國阿拉巴馬大學伯明翰分校病理學系呼吸道病原參考實驗室;中國生物技術(shù)股份有限公司;
【基金】：國家科技支撐計劃課題(2008BAI66B01) 國家“重大新藥創(chuàng)制”專項(2013ZX09402-302-215) 甘肅省科技重大專項(0801NKDA003) 甘肅省“十二五隴藥產(chǎn)業(yè)發(fā)展”專項
【分類號】：R378.15;R392-33

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