定量检测人黄体生成素的超顺磁性免疫层析法的初步建立  

Preliminary establishment of quantitative detection for luteinizing hormone based on superparamagnetic particles labeled immuno chromatography method

作　　者：王策[1] 梁臻龙[1] 管笛[3] 武会娟[3] 刘晓婷[2] 亢子佳[3] 张帆[2] 陈运霞[2] 刘萍[2] 杨梦回 刘雨桐[3] 王成彬[1] 

机构地区：[1]温州医科大学检验医学院生命科学学院,325035 [2]解放军总医院临床检验科 [3]北京市理化分析测试中心

出　　处：《中华医学杂志》2015年第44期3631-3634,共4页National Medical Journal of China

基　　金：北京市科学技术研究院青年骨干计划（201415）

摘　　要：目的应用超顺磁性磁珠标记的免疫层析技术建立用于黄体生成素（LH）快速定量检测的方法。方法采用NHS／EDC修饰磁珠，以1株LH单克隆抗体与磁珠偶联，另1株LH抗体包被于硝酸纤维素膜，建立双抗体夹心法与免疫层析相结合的定量检测方法，通过线性范围、精密度、准确度、特异性、稳定性等对所建方法进行性能评价。收集常规检测LH可信度较高的化学发光法（CLIA）检测后标本，再用所建方法重复检测，验证方法的可靠性。结果磁珠偶联LH抗体与LH结合后，与硝酸纤维素膜包被LH抗体反应磁信号检测最佳时间为20min；所建方法低值、中值、高值变异系数（Cv）为8％-12％；偏倚〈10％；回收率为90％～120％，灵敏度为0．63mlU／ml，与人绒毛膜促性腺激素（hCG）、促甲状腺激素（TSH）、促卵泡激素（FSH）无明显交叉反应，与CLIA比对检测结果，相关性良好（R2=0．96，P〈0．05）。结论超顺磁性磁珠标记免疫层析法检测血清LH简便快速、灵敏度高，有望成为即时检验（POCT）方法定量检测生物样品中微量成分发展的方向。Objective To develop a quick quantitative detecting method for luteinizing hormone （ LH ） based on superparamagnetic particles labeled immunochromatography technology. Methods Magnetic particles were catalyzed by EDC/NHS, LH monoclonal antibody were coupled with magnetic particles, another antibody were coated with the NC membrane, established a quantitative detecting method combined sandwish assay format with immunochromatography. The performance of this method was evaluated by linear range, precision, accuracy, specificity and stability. Detecting the serum sample that were tested by chemiluminescence immunoassay（CLIA） which was high credibility to verify the reliability. Results The reaction time of LH antibody coupled magnetic particles, LH and LH antibody coated in nitrocellulose membrane was 20 rain ; the coefficient of variation （CV） values for low, median, high were 8% - 12%, the bias was less than 10% , recovery rate was 90% - 120% ,the minimum detection limit was 0. 63 mlU/ml, no obvious cross reaction with human chorionic gonadotropin （HCG）, thyroid stimulating hormone （TSH）, follicle-stimulating hormone （FSH）. Test results of clinical sample had good correlation with CLIA （ R2 = 0. 96, P 〈 0. 05 ） . Conclusion The superparamagnetic particles labeled immuno-chromatography method is simple and rapid, and is expected to become a direction in the development for point-of-care test （POCT） quantitative detection of microcompnents in biological sample.

关 键 词：促黄体激素 超顺磁 免疫层析法 

分 类 号：R446.6[医药卫生—诊断学]