摘要目的 侵袭性真菌病病原学诊断的经典培养方法耗时长,常常导致临床确诊延误.本研究通过荧光染色技术对侵袭性真菌病尤其是少见真菌病进行快速诊断,探讨其临床应用价值.方法 回顾性分析我院2017年9月—2018年9月通过荧光染色法进行侵袭性真菌病菌种鉴定及病原学诊断的病例,对其中3例少见侵袭性真菌病进行病例总结报道.对临床标本进行预处理,直接涂片并滴上荧光染液,捻磨混匀后在荧光显微镜下直接观察,根据镜下菌体形态进行菌种鉴定.预处理后标本同时进行常规细菌及真菌培养,根据典型菌体形态初步鉴定菌种,运用PCR技术扩增及测序分析明确具体病原.结果 运用荧光染色检验技术检出3例临床少见病例.根据典型菌体荧光染色形态初步诊断分别为马尔尼菲蓝状菌(Talaromyces marneffei)、毛霉目(Mucorales)、无绿藻菌(Prototheca),测序技术结果提示为马尔尼菲蓝状菌、少根根霉菌、小型无绿藻菌.结论 荧光染色技术是一种快速、经济和直观的真菌诊断方法,经荧光染色后真菌轮廓清晰、易于识别,能对临床高度怀疑侵袭性真菌病进行提示性诊断,是少见或培养难以获得真菌病原学实验室诊断的重要补充.

abstractsObjective To evaluate the performance of an enhanced fluorescent staining for the rapid diagnosis of invasive mycosis, especially rare cases, considering the traditional culture method always leads to delays in clinical diagnosis for its time consuming. Methods Cases of invasive mycosis identified by fluorescent staining in our hospital from September, 2017 to September, 2018 were retrospectively analyzed. Three rare in-vasive infections were reported in this study. Clinical specimens were pretreated using standard procedures and then smeared on slides along with the enhanced fluorescent dye. Species of the pathogens were identified accord-ing to their morphology under fluorescent microscope. The traditional culture method was used as a standard method to identify the pathogenic species based on their colony morphology, followed by PCR and sequencing analysis for further confirmation. Results Three cases of invasive mycosis caused by rare pathogens of Talaro-myces marneffei, Mucorales and Prototheca were rapidly diagnosed with the fluorescent staining method. Sequen-cing results indicated the species were Talaromyces marneffei, Rhizopus arrhizus and Prototheca wickerhamii. Conclusions Fluorescent staining is a rapid, economic and direct method for the diagnosis of invasive mycosis. The morphology of fungi is clear and easy to identify after fluorescence staining, which could be used for indica-tive diagnosis of highly suspected invasive mycosis and serve as an important complement to the traditional cul-ture method, especially for the diagnosis of rare or uncultured fungal pathogens.