Myoken Yoshinari, Sugata Tatsumi, Mikami Yuzuru, Murayama Somay Yamagata, Fujita Yoshinori
Department of Oral Surgery, Hiroshima Red Cross and Atomic Bomb Survivors Hospital, Hiroshima, Japan.
J Oral Maxillofac Surg. 2008 Sep;66(9):1905-12. doi: 10.1016/j.joms.2008.01.053.
A definitive diagnosis of invasive oral aspergillosis can be difficult because the culturing of tissue samples frequently fails to isolate Aspergillus species. In addition, the mycelial elements of Aspergillus species seen in tissue sections are histopathologically indistinguishable from those of non-Aspergillus species. We analyzed the usefulness of a DNA probe directed against the alkaline proteinase (ALP) gene of Aspergillus fumigatus for the identification of Aspergillus species by the in situ hybridization (ISH) technique in patients with oral mycosis.
The ALP probe was tested on tissue specimens from 16 patients with hematologic malignancies who had invasive, orofacial fungal infections and a positive culture for one of the following organisms: Aspergillus species in 13 patients (A. flavus in 10, A. terreus in 2, and A. fumigatus in 1), and Exophiala dermatitis, Trichoderma longibrachiatum, and Candida albicans in 1 patient each. In situ hybridization with the ALP probe was performed using formalin-fixed, paraffin-embedded tissue samples.
The ALP probe showed a strong reaction with specimens from all 13 patients who had culture-proven aspergillosis specimens attributable to A. flavus, A. terreus, and A. fumigatus. On the other hand, the ALP probe showed no cross-reactivity with other fungi (Exophiala dermatitis, Trichoderma longibrachiatum, and Candida albicans).
These findings indicate that ISH using an ALP probe may increase the accuracy of diagnosing invasive oral aspergillosis in immunocompromised patients, and facilitate the provision of adequate antifungal treatment.
侵袭性口腔曲霉病的明确诊断可能存在困难，因为组织样本培养常常无法分离出曲霉菌种。此外，在组织切片中所见的曲霉菌种的菌丝成分在组织病理学上与非曲霉菌种的菌丝成分无法区分。我们分析了一种针对烟曲霉碱性蛋白酶（ALP）基因的DNA探针，通过原位杂交（ISH）技术在口腔真菌病患者中鉴定曲霉菌种的实用性。
对16例血液系统恶性肿瘤患者的组织标本进行了ALP探针检测，这些患者患有侵袭性口腔真菌感染，且以下其中一种微生物培养呈阳性：13例患者为曲霉菌种（10例为黄曲霉，2例为土曲霉，1例为烟曲霉），1例患者为皮炎外瓶霉，1例患者为长枝木霉，1例患者为白色念珠菌。使用福尔马林固定、石蜡包埋的组织样本进行ALP探针原位杂交。
ALP探针与所有13例经培养证实为黄曲霉、土曲霉和烟曲霉所致曲霉病标本的患者标本均呈现强烈反应。另一方面，ALP探针与其他真菌（皮炎外瓶霉、长枝木霉和白色念珠菌）无交叉反应。
这些发现表明，使用ALP探针进行ISH可能会提高免疫功能低下患者侵袭性口腔曲霉病的诊断准确性，并有助于提供适当的抗真菌治疗。
