改良荧光法测定血清涎酸及临床应用  

A modified fluorescent method and its clinical application for measurement of sialic acid in sera

作　　者：吴平江 黎明[1] 

机构地区：[1]广东医学院附属医院中心实验室

出　　处：《广东医学院学报》1998年第1期35-38,共4页Journal of Guangdong Medical College

摘　　要：目的：建立一种快速而灵敏的荧光分析法用于临床上血清涎酸分析测定。方法：首先用０．０５ｍｏｌ／Ｌ硫酸加热水解血清中脂结合态的涎酸（ＳＡ），再加入新的荧光反应试剂丙二腈，在０．１５ｍｏｌ／Ｌ、ｐＨ９．５的硼酸盐缓冲液中与ＳＡ结合反应，生成稳定的荧光产物，然后采用荧光分光光度计进行分析测定。结果：生成的荧光产物最大激发光波长为３８２ｎｍ，最大发射光波长为４４１ｎｍ，ＳＡ浓度在１．６２×１０－３～７．９×１０－７ｍｏｌ／Ｌ范围内与荧光强度呈良好线性关系（ｒ＝０．９９９５），最低检测限为２．４×１０－８ｍｏｌ／Ｌ，批内（ｎ＝２０）、批间（ｎ＝１０）测定变异系数分别为１．６５％和２．０５％。结论：本法与其它ＳＡ测定方法相比，具有更快速、简便、灵敏和特异性强等特点。Objective:To establish a rapid and highly sensitive fluorescent method suitably used for clinical detection of sialic acid in serum samples.Method:Firstly,serum sialic acid with lipid combinative state was released by heat hydrolysis in 0.05mol/L sulfate acid;secondly,sialic acid released was converted into a kind of highly fluorescent compounds in 0.15 mol/L borate buffer,pH9.5,with malononitrile;and finally,the fluorescent intensity of the reaction mixture was measured with a fluorospectrophotometer.Results:The excitation λ mam and emission λ mam of the stable fluorescent compound were 382nm and 441nm,respectively;the sialic acid concentration in the range of 1.62×10 3 to 7.9×10 7 mol/L was well linear related with the fluorescent intensity( r =0.9995);the minimum of detection is 2.4×10 8 mol/L;and within group( n =20) and between groups( n =10) coefficients of variation for measuring of sialic acid at the 7.9×10 7 mol/L level were 1.65% and 2.05%,respectively.Conclusion:The method for sialic acid measurement was simple,rapid,highly sensitive and adapted for clinical application.

关 键 词：涎酸 N-乙酰神经氨酸 丙二腈 荧光分析法 

分 类 号：R446.112[医药卫生—诊断学] Q592.1[医药卫生—临床医学]