University at Buffalo School of Medicine and Biomedical Sciences, Buffalo, NY, USA.
Otolaryngol Head Neck Surg. 2012 Nov;147(5):943-9. doi: 10.1177/0194599812451010. Epub 2012 Jun 11.
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.
研究胰蛋白酶预处理对慢性鼻-鼻窦炎(CRS)患者黏液中真菌的组织学检测的改良作用,方法:采用组织学方法,对 12 例 CRS 手术患者的 34 个鼻窦标本进行研究,对其进行苏木精和伊红、Gomori 六胺银(GMS)染色、GMS 联合胰蛋白酶消化、几丁质酶免疫荧光染色和 Alternaria 免疫荧光染色,所有患者均接受皮肤试验、总 IgE 血清学和 23 种真菌特异性 IgE 抗体的放射过敏原吸附试验(RAST)检测。结果:常规 GMS 染色仅在 34 个标本中的 9 个(27%)中检测到真菌。用胰蛋白酶预处理标本可显著提高真菌的可视化效果(31/34,91%)。几丁质酶免疫荧光在 34 个标本中的 32 个(94%)中显示真菌,抗 Alternaria 在 34 个标本中显示 33 个(97%)。仅 12 例患者中的 8 例(75%)有可检测的过敏症。结论:本报告描述了一种改良的常规 GMS 染色方法,该方法可显著提高组织学中真菌的可视化效果,并解释了之前研究中检测不到真菌的原因。这些新颖的、更敏感的组织学方法揭示了真菌存在于变应性和非变应性 CRS 患者的嗜酸性黏蛋白中,进一步质疑了 IgE 介导的病理生理学的关键作用。