人血清中A、C、Y、W135群脑膜炎球菌荚膜多糖IgG抗体含量ELISA检测方法的建立

发布时间：2019-03-18 10:51

【摘要】：目的建立人血清中抗A、C、Y、W135群脑膜炎球菌荚膜多糖Ig G抗体含量的ELISA定量检测方法,并进行验证。方法筛选适用的酶标板,优化抗原包被浓度,确定ELISA检测过程的关键条件,对建立的ELISA方法的特异性、线性、准确性、精密度、最低检测值和稳定性进行验证,并建立实验室质控血清的质控检测范围。结果酶标板的板内CV为9.8%,板间CV为16.4%,符合检测使用要求。免疫血清经多糖吸收后,群特异性抗体含量与同群多糖吸收剂量呈明显的剂量依赖关系。12份美国CDC质控血清中各群Ig G抗体含量与血清的稀释度呈负相关,相关系数(r)和斜率(slope)均接近-1,线性关系良好;检测结果与CDC公布数据一致性良好,一致性相关系数A、C、W135、Y群分别为0.912 5、0.939 6、0.963 7和0.920 6,准确度较好。实验室内部质控血清Men20中各群Ig G抗体含量试验内和试验间精密度分别小于10%和20%,精密度较好;各血清群别的CV值均30%,稳定性较好;A、C、Y、W135群多糖Ig G抗体含量最低检测值分别为0.092、0.118、0.067和0.035μg/ml,质控血清的检测范围分别为:92.49~36.53、37.29~17.69、66.18~25.54和50.15~23.31μg/ml。结论建立了标准化的检测人血清中抗A、C、Y、W135群脑膜炎球菌多糖Ig G抗体含量的定量ELISA方法,并确定了实验室日常用质控血清Men20的质控检测值范围。
[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]
【作者单位】：兰州生物制品研究所有限责任公司甘肃省疫苗工程技术研究中心;中国食品药品检定研究院;美国阿拉巴马大学伯明翰分校病理学系呼吸道病原参考实验室;中国生物技术股份有限公司;
【基金】：国家科技支撑计划课题(2008BAI66B01) 国家“重大新药创制”专项(2013ZX09402-302-215) 甘肃省科技重大专项(0801NKDA003) 甘肃省“十二五陇药产业发展”专项
【分类号】：R378.15;R392-33

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