title: Rapid and reliable diagnosis of Wilson disease using X-ray fluorescence.
abstract: Wilson's disease (WD) is a rare autosomal recessive disease due to mutations of the gene encoding the copper-transporter ATP7B. The diagnosis is hampered by the variability of symptoms induced by copper accumulation, the inconstancy of the pathognomonic signs and the absence of a reliable diagnostic test. We investigated the diagnostic potential of X-ray fluorescence (XRF) that allows quantitative analysis of multiple elements. Studies were performed on animal models using Wistar rats (n = 10) and Long Evans Cinnamon (LEC) rats (n = 11), and on human samples including normal livers (n = 10), alcohol cirrhosis (n = 8), haemochromatosis (n = 10), cholestasis (n = 6) and WD (n = 22). XRF experiments were first performed using synchrotron radiation to address the elemental composition at the cellular level. High-resolution mapping of tissue sections allowed measurement of the intensity and the distribution of copper, iron and zinc while preserving the morphology. Investigations were further conducted using a laboratory X-ray source for irradiating whole pieces of tissue. The sensitivity of XRF was highlighted by the discrimination of LEC rats from wild type even under a regimen using copper deficient food. XRF on whole formalin-fixed paraffin embedded needle biopsies allowed profiling of the elements in a few minutes. The intensity of copper related to iron and zinc significantly discriminated WD from other genetic or chronic liver diseases with 97.6% specificity and 100% sensitivity. This study established a definite diagnosis of Wilson's disease based on XRF. This rapid and versatile method can be easily implemented in a clinical setting.
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France.
authors: Slávka
authors: S
authors: Kaščáková
authors: Ligne de lumière NANOSCOPIUM, Synchrotron SOLEIL Gif sur Yvette, F-91192 France.
authors: Cameron M
authors: CM
authors: Kewish
authors: Laboratoire de Physique des Solides UMR CNRS 8502, Univ Paris-Sud, Bâtiment 510 Orsay Cedex 91405 France.
authors: Stéphan
authors: S
authors: Rouzière
authors: INSERM, Unité 1064, Centre de Recherche en Transplantation & Immunologie, ITUN, CHU Hôtel DieuNantesFrance; Univ d'AngersHIFIH, UPRES 3859AngersFrance.
authors: Françoise
authors: F
authors: Schmitt
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France; AP-HP Hôpital Paul Brousse, Centre Hépato-BiliaireVillejuifF-94800France; Centre de Référence National de la Maladie de Wilson, AP-HPFrance.
authors: Rodolphe
authors: R
authors: Sobesky
authors: Centre de Référence National de la Maladie de Wilson, AP-HPFrance; AP-HP Hôpital Lariboisière, Laboratoire de toxicologie biologiqueParis Cedex 1075475France.
authors: Joël
authors: J
authors: Poupon
authors: Ligne de lumière SMIS, Synchrotron SOLEIL Gif sur Yvette, F-91192 France.
authors: Christophe
authors: C
authors: Sandt
authors: Service de Génétique Moléculaire Pharmacogénétique et Hormonologie Hôpital Bicêtre, Le Kremlin-Bicêtre Cedex F-94276 France.
authors: Bruno
authors: B
authors: Francou
authors: Ligne de lumière NANOSCOPIUM, Synchrotron SOLEIL Gif sur Yvette, F-91192 France.
authors: Andrea
authors: A
authors: Somogyi
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France; AP-HP Hôpital Paul Brousse, Centre Hépato-BiliaireVillejuifF-94800France; Centre de Référence National de la Maladie de Wilson, AP-HPFrance.
authors: Didier
authors: D
authors: Samuel
authors: DHU HepatinovVillejuif, F-94800France; Centre de Référence National de la Maladie de Wilson, AP-HPFrance; Service d'Hépatologie et de Transplantation Hépatique Pédiatriques, Hôpital Bicêtre, AP-HP, Université Paris SudLe Kremlin Bicêtre Cedex, F-94275France; Univ Paris-SudUMR-S 1174OrsayF-91400France.
authors: Emmanuel
authors: E
authors: Jacquemin
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France.
authors: Anne
authors: A
authors: Dubart-Kupperschmitt
authors: INSERM, Unité 1064, Centre de Recherche en Transplantation & Immunologie, ITUN, CHU Hôtel Dieu Nantes France.
authors: Tuan Huy
authors: TH
authors: Nguyen
authors: Univ Paris 6, Sorbonne Universités, UPMC, Collège de France, Laboratoire de Chimie de la Matière Condensée de Paris (LCMCP) Paris F-75005 France.
authors: Dominique
authors: D
authors: Bazin
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France; AP-HP Hôpital Paul Brousse, Centre Hépato-BiliaireVillejuifF-94800France; Centre de Référence National de la Maladie de Wilson, AP-HPFrance.
authors: Jean-Charles
authors: JC
authors: Duclos-Vallée
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France; AP-HP Hôpital Paul Brousse, Service d'Anatomo-PathologieVillejuifF-94807France.
authors: Catherine
authors: C
authors: Guettier
authors: INSERMUnité 1193, Villejuif, F-94800France; Univ Paris-SudUMR-S 1193, Villejuif, F-94800France; DHU HepatinovVillejuif, F-94800France.
authors: François
authors: F
authors: Le Naour
keywords: Wilson disease; X‐ray fluorescence spectroscopy; copper; diagnosis