gms | German Medical Science

102. Jahrestagung der DOG

Deutsche Ophthalmologische Gesellschaft e. V.

23. bis 26.09.2004, Berlin

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Assessment of the use of H.O.P.E. fixation in vitrectomy specimens in patients with chronic uveitis (Masquerade syndrome)

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  • corresponding author S.E. Coupland - Department of Pathology, University Medicin, Campus Benjamin Franklin
  • H. Heimann - Department of Ophthalmology, University Medicine, Campus Benjamin Franklin
  • N. Bechrakis - Department of Ophthalmology, University Medicine, Campus Benjamin Franklin
  • A. Perez-Canto - Department of Pathology, University Medicin, Campus Benjamin Franklin
  • M. Hummel - Department of Pathology, University Medicin, Campus Benjamin Franklin
  • M.H. Foerster - Department of Ophthalmology, University Medicine, Campus Benjamin Franklin
  • H. Stein - Department of Pathology, University Medicin, Campus Benjamin Franklin

Evidenzbasierte Medizin - Anspruch und Wirklichkeit. 102. Jahrestagung der Deutschen Ophthalmologischen Gesellschaft. Berlin, 23.-26.09.2004. Düsseldorf, Köln: German Medical Science; 2004. Doc04dogFR.16.07

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/dog2004/04dog308.shtml

Published: September 22, 2004

© 2004 Coupland et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.

Outline

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Objective

Cytological examination of rapidly-transported, unfixed vitreal specimens is considered the gold standard in exclusion of primary intraocular lymphoma (PIOL) in patients with idiopathic steroid resistant chronic uveitis. These specimens are difficult, and reports of "false negatives" or "false positives" are common. Fixation using H.O.P.E. technique (Herpes-Glutamic acid buffer mediated Organic solvent Protection Effect) has been successfully applied in the investigation of cytospin preparations using immunohistochemistry, in situ hybridisation and polymerase chain reaction (PCR). H.O.P.E. fixation of vitrectomy specimens in patients with clinical reactive vitritis and PIOL was performed, to determine its usefulness with these biopsies.

Methods

Pars plana vitrectomy was performed in 12 cases of patients with chronic uveitis. The vitreous samples were halved: one half was fixed in H.O.P.E. solution, the other half remained unfixed. The specimens were assessed blindly by two pathologists, and the corresponding key catalogued by the operating surgeon. Furthermore, immunocytology and, when possible, PCR for the detection of immunoglobulin heavy chain gene rearrangement (IgH-PCR) and GeneScan was performed.

Results

11/12 vitreous samples were diagnosed as reactive vitritis; one sample, a primary intraocular lymphoma of B-cell type. Compared to the unfixed vitreal specimens, the quality of the cytomorphology and immunocytology improved in the H.O.P.E.-fixed specimens. IgH-PCR and GeneScan analysis sequencing demonstrated polyclonal amplification products in the reactive cases, and a monoclonal B-cell population in the B-PIOL.

Conclusions

Our results demonstrate that cytomorphology and immunoreactivity of vitreous specimens are well preserved following H.O.P.E.-fixation. DNA of sufficient quality could be extracted from H.O.P.E.-fixed vitreous specimens, in order to perform clonality analyses. H.O.P.E. fixation appears to be promising in improving the diagnostic reliability of vitreous specimens in patients with masquerade syndrome.