不同方法测定PKU随访患者干血滤纸片Phe浓度的比较  

Comparison of different methods for measuring the phenylalanine concentration of phenylketonuria patients in dried blood spots

作　　者：占霞 邱文娟[1] 汪培承 徐烽[1] 鲍培忠[1] 高晓岚[1] 叶军[1] 顾学范[1] Zhan Xia , Qiu Wenjuan, Wang Peicheng, Xu Feng, Bao Peizhong, Gao Xiaolan, Ye Jun, Gu Xuefan(Department of Pediatric Endocrinology and Genetics, Shanghai Institute for Pediatric Research, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, Chin)

机构地区：[1]上海交通大学医学院附属新华医院上海市儿科医学研究所小儿内分泌遗传科,200092 [2]珀金埃尔默医学诊断产品(上海)有限公司

出　　处：《中华检验医学杂志》2018年第5期361-365,共5页Chinese Journal of Laboratory Medicine

基　　金：国家重点研发计划精准医学研究专项（2016YFC0905100）;国家自然科学基金（30973216,81170811）;上海市卫生局基金（20134005,20164Y0171）

摘　　要：目的 比较串联质谱（MS/MS）,包括非衍生标准曲线定量法（简称＂标曲法＂）和衍生法、非衍生法及荧光法在苯丙酮尿症（PKU）随访患者血苯丙氨酸（Phe）浓度测定中的应用.方法 收集上海新华医院随访的204例PKU患者干血滤纸片（DBS）样本;MS/MS标曲法、衍生法、非衍生法及荧光法测定Phe浓度,分为低、中、高,即〈360、360-600和〉600 μmol/L 3组,对Phe检测值进行一致性检验及非参数检验.结果 MS/MS标曲法批内和批间精密度为4.0%-7.2%,回收率为93.2%-97.3%.一致性检验显示各组Phe检测值在95%一致性界限外的值不超过8.1%.低、中浓度组,荧光法、MS/MS标曲法和衍生法Phe检测值四分位数分别为176（118,251）μmol/L、174（94, 273）μmol/L、153（94,242）μmol/L,和540（478,578）μmol/L、485（414,529）μmol/L、466（402,513） μmol/L;荧光法较MS/MS标曲法高4.8%-9.0%,（Z=-3.787--2.674,P〈0.01）;MS/MS非衍生法较标曲法低3.9%-5.2%,（Z=-7.474--5.747,P〈0.01）;高浓度组,MS/MS标曲法、衍生法、非衍生法和荧光法检测值为807（695,924）μmol/L,700（575,785）μmol/L,680（623,771）μmol/L和711（674,794）μmol/L,MS/MS标曲法较其他方法高10.9%-16.2%,（Z=-4.458--4.356,P〈0.01）.结论 MS/MS标曲法测定DBS中Phe浓度特异性和准确度较好,4种方法测定血Phe浓度一致性较好,虽检测值存在差异,但均可用于多数PKU患者的Phe浓度监测.Objective To compare the phenylalanine（Phe）concentration in the sample of dried blood spot,measured by four different methods：fluorescence assay,tandem mass spectrometry（MS/MS）, including MS/MS Derivatized,MS/MS Non-Derivatized and MS/MS-Standard Curve.Methods A total of 204 dried blood spot（DBS）samples of phenylketonuria（PKU）patients from Shanghai Xinhua Hospital were collected in this study.Phe concentration in DBS was measured by fluorescence assay and MS /MS assay,including MS/MS Derivatized, MS/MS Non-Derivatized and MS/MS-Standard Curve.The samples were divided into low, middle and high concentration groups according to Phe concentration, which were under the 360 μmol/L,between 360 and 600 μmol/L,and over 600 μmol/L respectively.The differences among the groups were analyzed by consistency check and non-parametric test.Results The within-day and between-day precisions of MS/MS-Standard Curve assay were 4.0%-7.2%,the recoveries were 93.2%-97.3%.Consistency check showed that less than 8.1%of Phe value were out of the 95%limit of agreement among these four methods.In the low and middle concentration groups,the quartile of Phe value measured by fluorescence assay,MS/MS Standard Curve and MS/MS Derivatized were 176（118,251）μmol/L,174 （94,273）μmol/L,153（94,242）μmol/L; and 540（478,578）μmol/L,485（414,529）μmol/L,466（402,513）μmol/L,respectively.Phe value measured by fluorescence assay was significantly higher than that by MS/MS Standard Curve,the difference was 4.8%-9.0%,（Z=-3.787 to -2.674,P〈0.01）. Phe value obtained from MS/MS Non-Derivatized was less than that by MS/MS-Standard Curve, the difference was 3.9%-5.2%,（Z=-7.474 to -5.747,P〈0.01）.In the high concentration group,the quartile of Phe value measured by MS/MS-Standard Curve, MS/MS Derivatized, MS/MS Non-Derivatized and fluorescence assay were 807（695,924）μmol/L,700（575,785）μmol/L,680（623,771）μmol/L and 711（674, 794）μmol/L, respectively.Phe value obtained from MS/MS Standard Cu

关 键 词：苯丙酮尿症 苯丙氨酸 干燥血斑检测 串联质谱法 

分 类 号：R725.8[医药卫生—儿科]