摘要
【目的】建立一种快速、简便、可靠的尿粘多糖定量检测方法,用于临床筛查粘多糖病(mucopolysacchari-dosis,MPS)患者。【方法】采用1,9-二甲基亚甲蓝(1,9-dimethylmethylene blue,DMB)在碱性条件下与尿中粘多糖(glycosaminoglycan,GAG)反应生成粉红色产物及分光光度计比色原理,检测267例正常人、12例MPS患儿及30例非MPS患儿的尿GAG含量,并同时检测尿肌酐(creatinine,Cr),计算GAG/Cr比值。【结果】正常人尿GAG水平常低于70 mg/L,且与年龄无显著相关。而GAG/Cr比值随年龄的递增呈递减趋势。除1例IVA型MPS患者外,其余11例MPS患儿尿GAG浓度及GAG/Cr比值均显著高于同龄正常儿童参考值,而非MPS患儿的尿GAG及GAG/Cr比值均在正常范围内。GAG测定的批内变异系数为1.3%,批间变异系数为3.6%,硫酸类肝素的回收率为(102±6.0)%。【结论】DMB-Tris比色法测定尿GAG是一种快速、简便、标本用量少、特异性和敏感性均较高的定量检测方法,可作为MPS的筛查方法在临床上推广应用。
[Objective] To set up a quantitative screening procedure for mucopolysaccharidosis （MPS） by measuring urinary glycosaminoglycan（GAG） based on 1,9-dimethylmethylene blue（DMB）-Tris assay. [Methods] The urinary GAG were detected by spectrophotometry and compared with that of appropriate standard solutions of heparan sulfate in 267 normal controls aged from neonates to adults, 30 non-MPS patients and 12 patients with MPS diagnosed on the basis of clinical symptoms, abnormal urinary GAG electrophoresis and enzyme assay in leukocytes. The GAG was corrected for creatinine （Cr） content. [Results] The GAG concentrations were less than 70 mg/L in all normal controls and there was not significant relationship between the levels of GAG and ages. The ratios of GAG/Cr （mg/mmol） in normal controls were age-dependent from 39.1± 13.0 in neonates to 3.9 - 1.6 in adults. Urinary GAG concentrations and GAG/Cr in patients with MPS were increased significantly compared to agematched normal values except one patient with type IVA . Urinary GAGs excretions in all non-MPS patients were normal. The within-run CV and between-run CV were 1.3 % and 3.6 %, respectively. [Conclusions] This quantitative measurement of GAG based on DMB-Tris assay is a simple and reliable screening procedure for MPS. Normal reference values from a healthy population have been obtained and shown to be age dependent.
出处
《中国儿童保健杂志》
CAS
2010年第11期885-888,共4页
Chinese Journal of Child Health Care
基金
广东省自然科学基金资助项目(30307-2080032)
关键词
粘多糖病
粘多糖
1
9-二甲基亚甲蓝
比色法
mucopolysaccharidosis
glycosaminoglycan
1,9-dimethylmethylene-blue
spectrophotometry
