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  • Improving the detection of fungi in eosinophilic mucin: seeing what we could not see before.

Improving the detection of fungi in eosinophilic mucin: seeing what we could not see before.

Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery (2012-06-13)
Canting Guo, Saied Ghadersohi, Gail M Kephart, Roger A Laine, David A Sherris, Hirohita Kita, Jens U Ponikau
ABSTRACT

To investigate the improvement in histologic detection of fungi with Gomori methenamine silver (GMS) stain by trypsin predigestion in the mucus of patients with chronic rhinosinusitis (CRS). Prospective, single group, descriptive analysis. Multi-institutional. Thirty-four sinus specimens from 12 surgical patients with CRS were stained with hematoxylin and eosin, GMS stain, GMS with trypsin digestion, immunofluorescence stains for chitinase, and anti-Alternaria. All patients received skin testing, total IgE serology, and radioallergosorbent tests (RAST) for 23 fungal-specific IgE antibodies. The conventional GMS stain detected fungi in only 9 of 34 (27%) specimens. Predigesting the specimen with trypsin dramatically improved the visualization of fungi (31/34, 91%). The chitinase immunofluorescence visualized fungi in 32 of 34 (94%), and anti-Alternaria visualized 33 of 34 specimens (97%). Only 8 of 12 (75%) patients had detectable allergies. This report describes a simple modification of the conventional GMS stain that can significantly improve the visualization of fungi on histology and explains the lack of detection in previous studies. These novel, more sensitive histologic methods reveal the presence of fungi within the eosinophilic mucin in allergic and also nonallergic CRS patients, further questioning a crucial role of an IgE-mediated pathophysiology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), lyophilized powder
Sigma-Aldrich
Trypsin Inhibitor from Glycine max (soybean), Suitable for manufacturing of diagnostic kits and reagents, suitable for cell culture
Methenamine, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Hexamethylenetetramine, ReagentPlus®, 99%
Sigma-Aldrich
Hexamethylenetetramine, meets analytical specification of Ph. Eur., BP, 99-100.5% (calc. to the dried substance)
Sigma-Aldrich
Hexamethylenetetramine, puriss. p.a., reag. Ph. Eur., ≥99.5% (calc. to the dried substance)
Sigma-Aldrich
Hexamethylenetetramine, ACS reagent, ≥99.0%