免疫组织化学作为中美洲巴拿马皮肤利什曼病诊断有用工具的性能

Performance of immunohistochemistry as a useful tool for the diagnosis of cutaneous leishmaniasis in Panama, Central America.

作者信息

Gonzalez K, Calzada J E, Díaz R, Paz H, García V, Miranda A, Tomokane T, Puga S, Saldaña A, Laurenti M

机构信息

Instituto Conmemorativo Gorgas de los Estudios de la Salud (ICGES), Panama; Laboratório de Patologia de Moléstias Infecciosas, Faculdade de Medicina, Universidade de Sao Paulo, São Paulo, SP, Brazil.

Instituto Conmemorativo Gorgas de los Estudios de la Salud (ICGES), Panama.

出版信息

Parasitol Int. 2019 Aug;71:46-52. doi: 10.1016/j.parint.2019.03.007. Epub 2019 Mar 16.

DOI:10.1016/j.parint.2019.03.007

PMID:30885795

Abstract

Cutaneous leishmaniasis (CL) is one of the most frequent parasitic zoonoses in Panama. Currently, conventional, molecular and histopathological tests are performed to diagnose CL. Immunohistochemistry (IHC) has proven to be a valuable tool to facilitate the diagnosis of leishmaniasis and to study the cellular immune response developed during the infection. Therefore, considering the absence of IHC in the diagnostic routine in Panama, the objective of this study is to demonstrate the usefulness of this test as a complementary diagnostic tool for improving the sensitivity of histopathology (HP) and helping to study the cellular immune response of patients. Samples from patients with suspected CL were analysed by intradermal reaction of Montenegro (IDRM), smears, culture, PCR (Viannia, Hsp-70), HP and IHC. According to the diagnostic criteria, 95.8% of patients were positive for Leishmania sp., that was characterized as Leishmania (V.) panamensis by PCR-HSP70/RFLP. From positive samples, Leishmania was detected by the tested diagnostic methods in the following degrees: 100% by IDRM, 60% by smears, 93.3% by culture, 100% by kDNA PCR, 78.3% by PCR Hsp-70, 50% by HP and 73.9% by IHC. Although IHC had a poor correlation (k = 0.191) with the diagnostic criteria, the sensitivities of both HP (76.1%) and smears (89.1%) were improved by combining them with IHC. IHC considerably improved the detection of the Leishmania parasites in the histopathological sections, supporting the need to implement this diagnostic tool in Panama. In addition, immunohistochemistry allows evaluation of the patient's immune response and thus provides new guidelines for the treatment and control of CL in Panama.

摘要

皮肤利什曼病（CL）是巴拿马最常见的寄生虫人畜共患病之一。目前，采用传统、分子和组织病理学检测方法来诊断CL。免疫组织化学（IHC）已被证明是一种有价值的工具，有助于利什曼病的诊断，并研究感染期间产生的细胞免疫反应。因此，鉴于巴拿马诊断常规中缺乏IHC，本研究的目的是证明该检测作为一种辅助诊断工具的有用性，以提高组织病理学（HP）的敏感性，并有助于研究患者的细胞免疫反应。对疑似CL患者的样本进行了蒙氏皮内反应（IDRM）、涂片、培养、PCR（维氏亚属、热休克蛋白70）、HP和IHC分析。根据诊断标准，95.8%的患者利什曼原虫属阳性，通过PCR-HSP70/RFLP鉴定为巴拿马利什曼原虫（V.）。在阳性样本中，通过以下检测诊断方法检测到利什曼原虫的比例如下：IDRM为100%，涂片为60%，培养为93.3%，kDNA PCR为100%，PCR Hsp-70为78.3%，HP为50%，IHC为73.9%。尽管IHC与诊断标准的相关性较差（k = 0.191），但将HP（76.1%）和涂片（89.1%）与IHC联合使用可提高其敏感性。IHC显著提高了组织病理学切片中利什曼原虫的检测率，支持在巴拿马应用这种诊断工具。此外，免疫组织化学可以评估患者的免疫反应，从而为巴拿马CL的治疗和控制提供新的指导方针。