Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Am J Clin Pathol. 2013 Aug;140(2):203-8. doi: 10.1309/AJCPNSU2SDZD9WPW.
To describe and discuss the utility and potential pitfalls of ribosomal RNA locus sequencing for direct identification of invasive fungi from fresh and formalin-fixed, paraffin-embedded specimens.
DNA was extracted from fresh and formalin-fixed, paraffin-embedded tissue and subjected to real-time polymerase chain reaction (PCR) targeting ITS2 and D2 regions of fungal ribosomal RNA locus. Cycle sequencing was performed on PCR products, and the identity of sequences was determined using a public database.
Four clinical cases of invasive fungal infection are presented to illustrate the utility of DNA sequencing for determining etiology when microbiological culture is negative, for shortening the time to identification of slow-growing fungi, for guiding antifungal therapy, and for shedding light on the pathogenesis of disseminated fungal infection.
Fungal ribosomal RNA locus sequencing from fresh or formalin-fixed, paraffin-embedded specimens is a powerful tool for rapid and accurate diagnosis of patients with culture-negative or uncultured invasive mycosis.
描述并讨论核糖体 RNA 基因测序在直接鉴定新鲜和福尔马林固定、石蜡包埋标本中的侵袭性真菌方面的实用性和潜在陷阱。
从新鲜和福尔马林固定、石蜡包埋的组织中提取 DNA，并进行靶向真菌核糖体 RNA 基因 ITS2 和 D2 区的实时聚合酶链反应（PCR）。对 PCR 产物进行循环测序，并使用公共数据库确定序列的身份。
呈现了 4 个侵袭性真菌感染的临床病例，以说明当微生物培养为阴性时，DNA 测序在确定病因、缩短慢生长真菌鉴定时间、指导抗真菌治疗以及阐明播散性真菌感染发病机制方面的实用性。
从新鲜或福尔马林固定、石蜡包埋的标本中进行真菌核糖体 RNA 基因测序是一种快速、准确诊断培养阴性或未培养侵袭性真菌感染患者的有力工具。
