血清层粘连蛋白荧光免疫层析检测方法的建立及性能评价

建立一种荧光定量层粘连蛋白(LN)检测方法,定量检测人血清LN水平。将鼠抗LN抗体用划线机喷到硝酸纤维素膜表面作为检测线(T),将链霉素亲和素用划线机喷到硝酸纤维素膜表面作为质控线(C);用生物素荧光微球标记鼠抗LN抗体作为缓冲液。利用双抗体夹心法原理定量检测LN水平,对方法的线性、精密度、空白限、回收率、相关性等性能指标进行验证,并使用市售化学发光LN试剂盒比对。荧光免疫分辨检测LN方法的回收率为86.03%~97.50%,空白限为4.22 ng/mL,线性范围为20~800 ng/mL,相关系数(r)为0.9989;批内精密度为6.51%,批间精密度为6.23%。与市售LN试剂平行检测100份血清标本,有良好的相关性(Y=1.0219X-1.1720,r=0.9871)。本研究建立了LN荧光免疫检测方法,各项性能指标均达到临床检验要求,操作便捷、结果准确,可用于临床血清LN水平的检测。

Establishment and performance evaluation of a fluorescence immunochromatographic assay for detection of serum laminin

To establish a quantitative detection method for Laminin(LN)based on Time-Resolved Fluorescence Immunoassay.The mouse anti-LN antibody was sprayed onto the surface of the nitrocellulose membrane with a marking machine as the detection line(T).Streptomycin avidin was sprayed onto the surface of cellulose nitrate membrane by marking machine as the quality control line(C).Mouse anti LN antibody was labeled with biotin fluorescent microspheres as buffer.The LN was quantitatively detected by the double antibody sandwich method,the linearity,precision,limit of blank,recovery and correlation of the method were verified,and the commercial chemiluminescence LN kit was used for comparison.The recovery of the fluorescence immunoassay for LN was 86.03%-97.50%.The limit of blank of LN was 4.22 ng/mL,the linear range was 20-800 ng/mL,the correlation coefficient r was 0.9989,the within-run precision was 6.51%,and the between-run precision was 6.23%.Compared with the commercially available product(100 serum samples),LN showed the high consistency(Y=1.0219X-1.1720,r=0.9871).In this study,the fluorescence immunoassay detection method of LN was established.All the performance indexes met the requirements of clinical examination.It was easy to operate and accurate,and could be used for the detection of serum LN level in clinic.