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1、EFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for internal learning or discussion , forbidden for any other purpose EFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for inte
2、rnal learning or discussion , forbidden for any other purpose 2014 欧洲感染学会曲霉菌病治疗指南-诊断部分2014 ESCMID Aspergillus Guideline- Diagnostic GroupKatrien Lagrou, Dieter Buchheidt, Malcolm Richardson, Manuel Cuenca-Estrella, Chris Kibbler, Claus Peter Heussel, Markus Ruhnke, Mauritio Sanguinetti, Jrgen Lffler
3、, Catherine Beigelman- Aubry, Cornelia Lass-FlrlPresent by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in BarcelonaEFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for internal learning or discussion , forbidden for any other purpo
4、se Present by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in Barcelona治疗前临床分析标本采集 Pre-analytical clinical sample treatment患者人群 Population目的 Intention干预手段 InterventionSoRQoE备注 Comment所有患者 Any获取符合要求的 单纯粘液标本， 如痰液To achieve a homogenous sample of viscous samples such as sputum通过真菌溶解液使其液 化，如：胰腺液或淀
5、粉 酶Liquifaction using a mycolytic agent, eg. Pancreatin, sputolysinAIII必要检查；大量 的痰液培养（完 整标本）表现出 更高的阳性率通过肺泡灌洗液 或沉淀物的分析 以获得曲霉培养 的最佳标本 To achieve optimal recovery of Aspergillus from BAL by Centrifugation and investigation of the sediment肺泡灌洗液/支气 管吸引（最少10 分钟吸引超过 1000克标本） Centrifugation of BAL/bronchial
6、aspirates (1000g for at least 10 minutes)AIII联合PCR，能够 显著增加培养的 敏感率所有患者 Any通过真菌溶解液使其液 化，如：超声手段或二 硫代苏糖醇 Liquifaction using sonication And dithiothreitol必要检查；曲霉 分离取决于培养 的标本量；真菌 浓缩Fraczek et al. 2013, Baxter et al. 2011EFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal In
7、fectious Study GroupOnly for internal learning or discussion , forbidden for any other purpose Present by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in Barcelona直接镜检：荧光标定 Direct microscopy: Fluorescence brighteners患者人群 Population目的 Intention干预手段 InterventionSoRQoE备注 Comment所有患者 Any通过新鲜的临床 标本
8、进行真菌鉴 定To identify fungal elements in fresh clinical specimens (e.g. BALs)荧光染色剂的应用： Calcofluor white, Uvitex 2B,BlancophorApplication of fluorescent dyes: Calcofluor white, Uvitex 2B, BlancophorAIII必要检测；没有 对曲霉特异的检 测手段；高敏感 率；结果快速反 馈；广泛应用； 无法与其他酵母 菌进行区分；标 本的质量影响最 后的结果获得Lass-Flrl C. 2007, Rchel R. 1999
9、, Chander J. 1993EFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for internal learning or discussion , forbidden for any other purpose Present by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in Barcelona诊断工具：组织病理 Diagnostic tools H
10、istopathology患者人群 Population目的 Intention干预手段 InterventionSoRQoE备注 Comment所有患者 Any确定组织切片和 病变组织中的真 菌To identify fungal elements in histological sections and stains组织病理检查； HE 染色 GMS染色 PAS染色Histological examination Haematoxylin and eosin (HE); Gomoris methenamine silver stain (GMS); Periodic acid-Schiff
11、 (PAS);AIII组织学检查是基础检测；无法与其他酵母菌进行区 分； HE染色：困难； GMS染色：移除基地细胞；对丝状真菌更为敏感 PAS染色：有利于细胞计数和细胞细节检查；对曲 霉非特异但具有较高的敏感性；形态学可以确定真 菌的种类（毛霉菌，菌株分枝间的夹角呈90度，芽 孢）;快速检测；应用广泛；在130个中枢感染案例 中73例涉及曲霉菌病（56%）；在组织检查阳性的 案例中只有25%的培养阳性率Denning D. 2003, Vyzantiadis TA. 2012, Sundaram et al. 2006, Rchel R. 1999, Lass-Flrl C. 2007, C
12、hai JK. 2004, Kaufman L. 1997, Verweij PE. 1996, Hayden RT. 2003荧光染色： Calcofluor white, Uvitex 2B,Blancophor免疫组化 单克隆抗体WF-AF-1 或EB-A1 原位杂交 Immunohistochemistry Monoclonal antibody WFAF- 1 or EB-A1 In situ hybridisationAIIBII可应用于冷冻标本或石蜡包裹的组织具有潜在的价值-提供菌株的特异性数据和基因数据 ； 可获得的商品化单克隆抗体：WF-AF-1是对烟曲霉 ，黄曲霉和黑曲霉特
13、异性的抗体EFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for internal learning or discussion , forbidden for any other purpose Present by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in Barcelona临床标本培养 Culture of clinical samples患者人群 P
14、opulation目的 Intention干预手段 InterventionSoRQoE备注 Comment所有患者 Any从深部感染部位 分离菌株（活检 组织，血液，脑 脊液等）Primary isolation from deep sites (biopsies, blood, CSF) AIII必要检测；血液 抑制孢子的繁殖 ；BHI能够帮助分 离孢子再生；从 标本中重复分离 获得一些菌落或 同一真菌具有更 大的临床意义。定量培养不能有 效区分感染和定 植。无菌部位分离真 菌：确定种属Cuenca-Estrella 2011, Richardson HM 2000从非无菌标本中 分离菌株
15、（痰液 ，呼吸道吸引物 ，皮肤等）Primary isolation from non-sterile samples (sputum, respiratory aspirates, skin)在沙保弱葡萄糖琼脂、 马铃薯琼脂、脑心浸膏 琼脂中以at 30C 和 37C 培养72小时（特 殊培养基） Culture on SDA, BHI agar, PDA at 30C and 37C for 72 (specific media)在沙保弱葡萄糖琼脂、 马铃薯琼脂、脑心浸膏 琼脂中+ 庆大霉素和 氯霉素以at 30C 和 37C 培养72小时（特 殊培养基）Culture on SDA, B
16、HI agar, PDA with gentamicin PLUS chloramphenicol at 30C and 37C for 72 hEFISGEuropean Society of Clinical Microbiology and Infection Diseases ESCMID Fungal Infectious Study GroupOnly for internal learning or discussion , forbidden for any other purpose Present by Cornelia Lass-Florl Australia ECCMID 10th May 2015 in Barcelona人群/检测：培养阳性- 质谱鉴定MALDI-TOF Population/Test: Positive culture - MALDI-TOF患者人群 Population目的 Intention干预手段 Interven
