Fluorescence In Situ Hybridization for Diagnosis of Whipple's Disease in Formalin-Fixed Paraffin-Embedded Tissue.

Peter Braubach,Mark Philipp Kühnel,Hans-Heinrich Kreipe,Jean-Christophe Lagier,Didier Raoult,Florian Peter Länger,Danny Jonigk,Ioannis Anagnostopoulos,Steffen Zender,Torsten Lippmann

doi:10.3389/fmed.2017.00087

Abstract

Whipple’s disease (WD) is a rare chronic systemic infection with a wide range of clinical symptoms, routinely diagnosed in biopsies from the small intestine and other tissues by periodic acid–Schiff (PAS) diastase staining and immunohistological analysis with specific antibodies. The aim of our study was to improve the pathological diagnosis of WD. Therefore, we analyzed the potential of fluorescence in situ hybridization (FISH) for diagnosing WD, using a Tropheryma (T.) whipplei-specific probe. 19 formalin-fixed paraffin-embedded (FFPE) duodenal biopsy specimens of 12 patients with treated (6/12) and untreated (6/12) WD were retrospectively examined using PAS diastase staining, immunohistochemistry, and FISH. 20 biopsy specimens with normal intestinal mucosa, Helicobacter pylori, or mycobacterial infection, respectively, served as controls. We successfully detected T. whipplei in tissue biopsies with a sensitivity of 83% in untreated (5/6) and 40% in treated (4/10) cases of WD. In our study, we show that FISH-based diagnosis of individual vital T. whipplei in FFPE specimens is feasible and can be considered as ancillary diagnostic tool for the diagnosis of WD in FFPE material. We show that FISH not only detect active WD but also be helpful as an indicator for the efficiency of antibiotic treatment and for detection of recurrence of disease when the signal of PAS diastase and immunohistochemistry lags behind the recurrence of disease, especially if the clinical course of the patient and antimicrobial treatment is considered.

Highlights

Epidemiology, Clinical Presentation, and TherapyWhipple’s disease (WD) is a rare disease with an approximate prevalence of less than three cases per million people investigated in the general European population [1, 2]
Classification of macrophages: macrophages were classified with 1–3 regarding to the morphological characterization of periodic acid–Schiff diastase-positive macrophages described by von Herbay et al [16]. (Previous) therapy of Whipple’s disease (WD): “no” indicates that no specific WD therapy has started at time of biopsy, “yes” indicates that a specific antibiotic WD therapy has already been started at time of biopsy
Specimens of all 16 cases with WD were positive for T. whipplei in immunohistochemical analysis, using the polyclonal anti-T. whipplei antibody described by Lepidi et al [30] (Table 2)

Summary

Introduction

Whipple’s disease (WD) is a rare disease with an approximate prevalence of less than three cases per million people investigated in the general European population (data from Italy) [1, 2]. Related infectious organisms include Mycobacterium and Corynebacterium species [3]. T. whipplei are non-motile and rod-shaped, about 1.4 μm in length and surrounded by a trilaminar cell wall containing glycans at the external membrane [4]. WD is a multisystemic disease with manifestation in various extra-intestinal organs including the central nervous system (CNS) (10–50% of WD patients), the heart (endocarditis), joints (isolated arthritis, spondylodiscitis), the eye (uveitis), and lung (pneumonia) [7]

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