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免疫组织化学在犬巴西利什曼原虫(维安尼亚种)所致美洲皮肤利什曼病确诊中的更高敏感性:一种优化免疫组织化学方法的描述

Higher sensitivity of immunohistochemistry for bona fide diagnosis of dog Leishmania (Viannia) braziliensis-driven American tegumentary leishmaniasis: description of an optimized immunohistochemistry method.

作者信息

dos Santos Isabele Barbieri, Tortelly Rogerio, Quintella Leonardo Pereira, de Fátima Madeira Maria, Monteiro de Miranda Luisa Helena, Borges Figueiredo Fabiano, Carvalhaes de Oliveira Raquel de Vasconcellos, Pacheco Schubach Tânia Maria

机构信息

Programa de Epidemiologia e Vigilância em Saúde (PEPIVS), FIOCRUZ BRASILIA, Avenida L3 Norte, s-n, campus Universitario Darcy Ribeiro, Gleba A, CEP: 70910-000-Brasilia-DF

Serviço de Anatomia Patológica Doutor Jefferson Andrade dos Santos - Faculdade de Veterinária - Universidade Federal Fluminense, Niterói, RJ, Brazil.

出版信息

Trans R Soc Trop Med Hyg. 2015 Jul;109(7):469-76. doi: 10.1093/trstmh/trv034. Epub 2015 May 14.

Abstract

BACKGROUND

The in situ detection of parasite antigens in tissue sections by immunohistochemistry (IHC) is a diagnostic alternative for human American tegumentary leishmaniasis (ATL), but has not been used for the diagnosis of cutaneous lesions in dogs with ATL. This study describes the results of IHC for the detection of amastigote forms and other Leishmania sp. antigen-positive cells and compares the results of IHC, histopathology and cytopathology for the diagnosis of canine ATL. In addition, possible cross-reactivity with sporotrichosis is analyzed.

METHODS

Forty paraffin-embedded biopsies and 40 smears of cutaneous lesions from dogs with ATL, confirmed by isolation and characterization of Leishmania (Viannia) braziliensis, and 40 paraffin-embedded biopsies of cutaneous lesions from dogs with sporotrichosis, confirmed by isolation of Sporothrix schenckii in culture (control group), were studied.

RESULTS

Immunohistochemistry was more sensitive in detecting amastigote forms than cytopathology and histopathology, with a positivity rate of 70% (n=28) versus 37.5% and 22.5% for histopathology and cytopathology, respectively. Cytoplasmic staining of mononuclear and endothelial cells was detected by IHC, which was highly specific since no cytoplasmic staining of these cells or staining of fungal structures was observed in sporotrichosis fragments.

CONCLUSIONS

In view of the higher sensitivity of IHC in detecting Leishmania sp. antigen and patterns of positivity for Leishmania sp. antigen compared to histopathology or cytopathology and the absence of cross-reactions with sporotrichosis, we recommend this technique for the diagnosis of canine tegumentary leishmaniasis.

摘要

背景

通过免疫组织化学(IHC)在组织切片中对寄生虫抗原进行原位检测是人类美洲皮肤利什曼病(ATL)的一种诊断替代方法,但尚未用于诊断患有ATL的犬的皮肤病变。本研究描述了用于检测无鞭毛体形式和其他利什曼原虫属抗原阳性细胞的IHC结果,并比较了IHC、组织病理学和细胞病理学对犬ATL的诊断结果。此外,还分析了与孢子丝菌病可能的交叉反应。

方法

研究了40例经巴西利什曼原虫(维安尼亚利什曼原虫)分离和鉴定确诊患有ATL的犬的皮肤病变石蜡包埋活检组织和40份涂片,以及40例经申克孢子丝菌培养分离确诊患有孢子丝菌病的犬的皮肤病变石蜡包埋活检组织(对照组)。

结果

免疫组织化学在检测无鞭毛体形式方面比细胞病理学和组织病理学更敏感,阳性率分别为70%(n = 28),而组织病理学和细胞病理学的阳性率分别为37.5%和22.5%。通过免疫组织化学检测到单核细胞和内皮细胞的细胞质染色,这具有高度特异性,因为在孢子丝菌病切片中未观察到这些细胞的细胞质染色或真菌结构的染色。

结论

鉴于免疫组织化学在检测利什曼原虫属抗原方面比组织病理学或细胞病理学具有更高的敏感性,以及利什曼原虫属抗原的阳性模式,并且与孢子丝菌病无交叉反应,我们推荐该技术用于诊断犬皮肤利什曼病。

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