Gosepath Jan, Brieger Juergen, Vlachtsis Konstantin, Mann Wolf J
Department of Otolaryngology, Head and Neck Surgery, University of Mainz, School of Medicine, Mainz, Germany.
Am J Rhinol. 2004 Jan-Feb;18(1):9-13.
It has been postulated that fungal organisms might represent the immunologic target initiating and maintaining the disease process in patients with chronic rhinosinusitis (CRS). The presence of fungi in nasal mucus has been established by different groups, but so far it has not been shown how the immune system could even recognize such extramucosal--extracorporal--fungal targets. The aim of this study was to determine whether fungal DNA is present in tissue specimens taken from patients with polypoid CRS.
Twenty-seven surgical specimens were collected from patients suffering from CRS. Fifteen surgical specimens from healthy ethmoidal mucosa served as controls. A second set of controls consisted of five surgical specimens of acoustic neuroma, which were included to rule out contamination within the protocol. All paranasal tissue samples were treated and rinsed carefully with a solution of Dithiothreitol to digest any nasal mucus and ensure that only tissue was examined. A highly sensitive two-step polymerase chain reaction (PCR) was applied to detect fungal DNA, using one universal primer for unspecific detection of fungal DNA and a second primer pair specific for Alternaria.
Fungal DNA was detected in all 27 CRS specimens equally with both PCR primers. Controls from healthy paranasal mucosa were positive using the panfungal primers in 10 of 15 cases but were all negative for Alternaria DNA. PCR was negative for fungal DNA in all five neuroma specimens.
Fungal DNA can be detected within sinonasal tissue specimens of patients suffering from CRS. These findings need to be discussed with respect to the proposed hypothesis of the immune system recognizing extramucosal organisms and initiating an immune response in sensitized patients.
据推测，真菌生物体可能是引发和维持慢性鼻-鼻窦炎（CRS）患者疾病进程的免疫靶点。不同研究团队已证实鼻黏液中存在真菌，但迄今为止，尚未明确免疫系统如何识别此类黏膜外——体外——真菌靶点。本研究旨在确定息肉样CRS患者的组织标本中是否存在真菌DNA。
收集了27例CRS患者的手术标本。15例健康筛窦黏膜的手术标本作为对照。第二组对照包括5例听神经瘤手术标本，纳入该组对照是为了排除实验方案中的污染因素。所有鼻旁组织样本均用二硫苏糖醇溶液仔细处理和冲洗，以消化任何鼻黏液，确保仅对组织进行检测。采用高灵敏度的两步聚合酶链反应（PCR）检测真菌DNA，使用一对通用引物非特异性检测真菌DNA，另一对引物对链格孢属特异性识别。
两种PCR引物在所有27例CRS标本中均检测到真菌DNA。健康鼻旁黏膜对照中，15例中有10例使用泛真菌引物呈阳性，但链格孢属DNA检测均为阴性。所有5例神经瘤标本的真菌DNA PCR检测均为阴性。
在CRS患者的鼻窦组织标本中可检测到真菌DNA。对于免疫系统识别黏膜外生物体并在致敏患者中引发免疫反应这一假说，需要结合这些发现进行讨论了。
