目的 建立一种能够定量检测人降钙素原的荧光微球免疫层析方法,用于诊断细菌感染,并对试纸条进行性能评估.方法 采用荧光微球标记技术与免疫层析技术相结合的方式,依据双抗体夹心原理,制备降钙素原荧光微球免疫层析试纸条;并从灵敏度、特异性、抗干扰性、精密度、回收率、稳定性以及与Roche试剂盒的符合率等方面进行全面的方法学评价.结果 试纸条的灵敏度为0.05 ng/mL;具有高的特异性和抗干扰性;批间和批内变异系数均<10.93%;回收率在95%~105%;试剂盒的有效期>12个月;且与Roche降钙素原试剂盒(电化学发光法)的相关性良好,回归方程为Y=0.8709X+0.8549,r=0.9519.结论 PCT荧光微球免疫层析试纸条能够实现定量、灵敏、准确检测,满足临床检测PCT的需求.
Objective This study was designed to establish a quantitative detection method for procalcitonin based on fluorescent microspheres immunochromatography,which can be used in the diagnosis of bacterial infection.Methods Based on the double antibody sandwich method,fluorescence microspheres labeling technology and immunochromatographic technology were combined together to prepare PCT fluorescent microsphere immunochromatographic strips.Besides,the performance of the strip was roundly evaluated by the sensitivity,anti-interference performance,precision,recovery rate,stability,and its comparison with Roche detecting kits.Results The sensitivity of the strip was 0.05 ng/mL.The specificity and anti-interference performance were both well.The inter-assay and intra-assay variations were both less than 10.93%.The recovery rate ranged from 95% to 105%.The strip was valid for more than 12 months.Furthermore, compared to Roche detecting kits,the result displayed a high correlation.The regression equation was Y =0.8709X+0.8549,r =0.9519.Conclusion The PCT fluorescent micro-spheres immunochromato graphic strip could achieve highly quantitative,accurate,and rapid detection,which could satisfy the needs of the clinical diagnostic.