元素标记免疫分析联合电感耦合等离子体质谱技术在HCG检测中的应用  

Application of element-tagged immunoassay combined with inductively coupled plasma mass spectrometry in HCG detection

作　　者：蒲丹娜 姜文灿 温新宇[2] 郭广宏[2] 李萍[1] 王成彬[2] Pu Danna;Jiang Wencan;Wen Xinyu;Guo Guanghong;Li Ping;Wang Chengbin(Hebei North University,Zhangjiakou 075000,China;Center of Laboratory Medicine,the 1st Medical Center of PLA General Hospital,Beijing 100853,China)

机构地区：[1]河北北方学院,张家口075000 [2]解放军总医院第一医学中心医学检验中心,北京100853

出　　处：《中华检验医学杂志》2020年第3期284-290,共7页Chinese Journal of Laboratory Medicine

摘　　要：目的:使用元素标记免疫分析联合电感耦合等离子体质谱(ICP-MS)技术的免疫检测分析方法建立用于的人绒毛膜促性腺激素(HCG)的定量检测方法。方法:方法性能评价。以稀土元素Sm为标记物,建立双抗体夹心法的HCG定量检测体系,对其生物素化抗体用量、反应时间进行优化;参照美国临床实验室标准化协会(CLSI)EP系列文件和相关标准,通过检测限、线性范围、批内和批间精密度、准确度、交叉反应、干扰试验等指标评价所建立的检测体系的分析性能。之后,收集88例临床用电化学发光法(ECLIA)检测过HCG的血清标本,使用所建方法进行检测,检测结果与ECLIA方法检测结果进行相关性分析。结果:优化后整个检测可在30 min内完成,最佳的生物素化抗体用量为0.5μl;所建立体系的空白检出限(LOB)为0.29 mIU/ml,在1.16~8365.62 mIU/ml范围内线性良好,线性相关系数大于0.995(R^2=0.9980),回收率为97.53%~102.01%,高浓度样本与低浓度样本的批内和批间变异系数均小于10%,与促甲状腺激素(TSH)、黄体生成素(LH)、卵泡刺激素(FSH)无明显的交叉反应,不同浓度干扰物质引起的干扰偏倚均小于10%;所建立检测体系的检测结果与ECLIA的检测结果进行比对,相关性良好(R2=0.9600)。结论:元素标记免疫分析联合ICP-MS技术的免疫分析方法检测HCG具有精密度高、特异性强、准确度好等优点,其方法确认和性能验证结果满足临床要求,为该方法的临床应用提供了实验依据。Objective To establish an inductively coupled plasma mass spectrometry(ICP-MS)based immunoassay method for the quantitative detection of human chorionic gonadotropin(HCG),and evaluate the clinical applicability of this method.Methods Sm was selected as element tags,and the HCG quantitative detection system was established by double antibody sandwich method.The dosage of biotinylated antibody and reaction time were optimized.According to EP documents of Clinical and Laboratory Standards Institute(CLSI)and related standards,the analytical performance was evaluated after the establishment of the assay,including the limit of blank(LOB),linearity,precision,recovery,cross reactivity and interference test.And 88 clinical samples were measured using the new method compared to the electrochemical immunoassay(ECLIA)method.Results Total process completed within 30 min after optimization,and the optimal biotinylated antibody dosage was 0.5μl.The LOB was 0.29 mIU/ml.The linearity was good within the range of 1.16-8365.62 mIU/ml with the linear correlation coefficient greater than 0.995(R^2=0.9980),the recovery was 97.53%-102.01%.Both intra-and inter-assay coefficients of variation of the high-value sample and the low-value sample were less than 10%.And there was no significant cross-reaction with Luteinizing hormone(LH),follicular stimulating hormone(FSH),and thyroid-stimulating hormone(TSH).The interference bias caused by different concentrations of interference substances was less than 10%.When compared with the ECLIA method for clinical sample detection,the proposed method showed a significant correlation(R2=0.9600).Conclusion The proposed ICP-MS base immunoassay for HCG detection has good accuracy,high sensitivity and specificity,and the results of analytical performance verification meet the clinical requirements,which provides experimental basis for the clinical application of this method.

关 键 词：绒毛膜促性腺激素 免疫测定 分光光度法 原子 质谱分析法 

分 类 号：R446[医药卫生—诊断学] O65[医药卫生—临床医学]