快速檢驗法Fluconazole disc diffusion method 應用在臨床血流感染念珠菌鑑定之探討

Fluconazole disk diffusion method application on primary identification of species from candemia isolate

蕭舜鴻(Shun-Hung Hsiao)

指導教授 ： 蘇慶華

臺北醫學大學/醫學院/醫學科學研究所/碩士(2012年)

https://doi.org/10.6831/TMU.2012.00177

摘要

念珠菌屬（Candida）為最常見的真菌病原菌，由於不同種的念珠菌對於藥物的敏感性不同，臨床上在第一時間鑑定出念珠菌種是很重要的課題，目前臨床主要利用CHROMagarＴＭ-candida鑑定，CHROMagarＴＭ-candida利用各種念珠菌生長菌落的顏色差異，當作鑑定依據，當CHROMagar的顏色結果混淆時，會再進一步做API32C生化鑑定，但是API32C生化鑑定成本較高，需要病患自行負擔費用。先前研究發現白色念珠菌在抗真菌藥物fluconazole所產生的抑制圈中，可以觀察到豆莢（Peapod）細胞型態以及經由Phloxine B染成紅色之菌落，本實驗目的是想了解在其他臨床常見的念珠菌屬中是否也有上述的現象，並進一步當作鑑別的依據。本實驗收集目前臨床常見的前五種血液感染之念珠菌菌株Candida albicans、C. glabrata、C. parapsilosis、C. tropicalis、C. Krusei，利用含有Phloxine B的培養基搭配7 ?慊/per、35?n?慊/per兩種不同濃度的fluconazole disc來觀察抑制圈內細胞以及菌落的型態特徵，實驗結果發現除了C. albican以外其他菌種皆無發現豆莢（Peapod）狀的細胞型態。在7 ?慊/per 及35?n?慊/per fluconazole disc 下C. albicans、C. parapsilosis、C. tropicalis有抑制圈，在抑制圈內皆可觀察到phloxine B染色之菌落，其中C. albicans、C. parapsilosis有clear zone；C. tropicalis則沒有。C. glabrata、C. krusei只有在35?n?慊/per fluconazole disc下才有抑制圈的產生，其中只有C. glabrata在抑制圈的菌落可以被Phloxine B染成紅色。結果顯示此方法可以初步鑑別出C. albicans、C. parapsilosis、C. tropicalis、C. glabrata與C. kresei。本研究可以解決在使用CHROMagarＴＭ-candida時所出現顏色混淆的問題，提高臨床鑑定之準確性；在成本方面，本實驗方法的成本遠低於目前臨床鑑定念珠菌的使用方法。未來可能應用在臨床鑑定念珠菌屬上。

關鍵字

念珠菌 ； 氟康唑

並列摘要

Candida species were known being most of the causative fungi of invasive fungal infections. In clinical laboratory work, there were mainly two methods to check whether it was Candida albicans or non-albicans candida species. We can use germ tube assay or CHROMagarTM-Candida. In our previous study, we found that the frequency of white and opaque colony switch had raised to 1.7x10-1(in nature, below 10-4) in the cloudy zone (inhibitory zone) of the treatment with fluconazole and terbinafine. Opaque colony will change color from white to red(or pink) when it grows on the phloxine B containing medium. In the present study, we used 5 most commom clinical species: Candida albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei. After 7 ?慊/per and 35?n?慊/per two different concentration of fluconazole disc treatment, we found out that only C. albicans, C. parapsilosis, C. tropicalis were inhibited by 7 ?慊/per fluconazole disc and produced red(or pink) ring along the inhibitory zone. C. glabrata and C. krusei were only inhibited by 35?n?慊/per fluconazole disc, with only C. glabrata produced red(or pink) ring along the inhibitory zone. In addition, we also found that only C. albicans formed Peapod-like structure in the cloudy zone. These findings suggest the possibility of creating a new convenient method for primary identification of Candida by using fluconazole disk on Phloxine B-containing
YEPD mediumagar.

並列關鍵字

Candida ； Fluconazole disc diffusion

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