Validation of amino-acids measurement in dried blood spot by FIA-MS/MS for PKU management

被引：9

作者：

Bruno, C. ^{[1
,2
]}

Dufour-Rainfray, D. ^{[2
,3
]}

Patin, F. ^{[1
,2
]}

Vourch, P. ^{[1
,2
]}

Guilloteau, D. ^{[2
,3
]}

Maillot, F. ^{[4
,5
,6
]}

Labarthe, F. ^{[5
,6
,7
]}

Tardieu, M. ^{[5
,6
,7
]}

Andres, C. R. ^{[1
,2
,7
]}

Emond, P. ^{[2
,3
]}

Blasco, H. ^{[1
,2
,7
]}

机构：

[1] CHRU Tours, Lab Biochim & Biol Mol, Tours, France

[2] Univ Tours, UMR INSERM U930, F-37041 Tours, France

[3] CHRU Tours, Lab Med Nucl In Vitro, Tours, France

[4] CHRU Tours, Serv Med Interne, Tours, France

[5] Univ Tours, UMR INSERM U1069, F-37041 Tours, France

[6] Ctr Competences Malad Hereditaires Metab Enfant &, Tours, France

[7] CHRU Tours, Serv Med Pediat, Tours, France

来源：

CLINICAL BIOCHEMISTRY | 2016年 / 49卷 / 13-14期

关键词：

Mass spectrometry; Method validation; Phenylketonuria; Phenylalanine; TANDEM MASS-SPECTROMETRY; PHENYLKETONURIA; PHENYLALANINE; PATHOPHYSIOLOGY;

D O I：

10.1016/j.clinbiochem.2016.07.008

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Objectives: Phenylketonuria (PKU) is a metabolic disorder leading to high concentrations of phenylalanine (Phe) and low concentrations of tyrosine (Tyr) in blood and brain that may be neurotoxic. This disease requires a regular monitoring of plasma Phe and Tyr as well as branched-chain amino-acids concentrations to adapt the Phe-restricted diet and other therapy that may be prescribed in PKU. We validated a Flow Injection Analysis tandem Mass Spectrometry (FIA-MS/MS) to replace the enzymatic method routinely used for neonatal screening in order to monitor in parallel to Phe, Tyr and branched-chain amino-acids not detected by the enzymatic method. Design and methods: We ascertained the performances of the method: linearity, detection and quantification limits, contamination index, accuracy. We cross validated the FIA-MS/MS and enzymatic methods and we evaluated our own reference ranges to monitor Phe, Tyr, Leu, Val on 59 dried blood spots of normal controls. We also evaluated Tyr, Leu and Val concentrations in PKU patients to detect some potential abnormalities, not evaluated by the enzymatic method. Results: We developed a rapid method with excellent performances including precision and accuracy <15%. We noted an excellent correlation of Phe concentrations between FIA-MS/MS and enzymatic methods (p < 0.0001) based on our database which are similar to references ranges published. We observed that 50% of PKU patients had lower concentrations of Tyr, Leu and/or Val that could not be detected by the enzymatic method. Conclusion: Based on laboratory accreditation recommendations, we validated a robust, rapid and reliable FIA-MS/MS method to monitor plasma Phe concentrations but also Tyr, Leu and Val concentrations, suitable for PKU management. We evaluated our own reference ranges of concentration for a routine application of this method. (C) 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

引用

页码：1047 / 1050

页数：4

共 18 条

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