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[用于对从血培养中分离出的念珠菌属菌株进行抗真菌药敏试验的肉汤微量稀释法与E-test法的比较]

[Comparison of broth microdilution and E-test methods for the antifungal susceptibility testing of Candida spp. strains isolated from blood cultures].

作者信息

Ozcan Sema Keçeli, Mutlu Birsen, Dündar Devrim, Willke Ayşe

机构信息

Kocaeli Universitesi Tip Fakültesi, Tibbi Mikrobiyoloji Anabilim Dali, Kocaeli.

出版信息

Mikrobiyol Bul. 2010 Apr;44(2):263-71.

Abstract

The incidence of serious fungal infections, particularly invasive Candida infections exhibit an increasing trend in the last decades since the number of patients receiving immunosuppressive treatment is increasing. This situation eventually results in an increment in resistance to antifungal agents. The aim of this study was to compare the standard broth microdilution (BMD) and E-test methods for antifungal susceptibility testing of Candida species isolated from blood cultures in our hospital, against fluconazole, voriconazole, caspofungin and amphotericin B. A total of 46 Candida strains isolated from the blood cultures by BACTEC 9000 (Becton Dickinson, USA) and identified by conventional techniques and API 20C AUX (BioMerieux, France) during January 2006-December 2007, were included into this study. The identification results of the isolates were as follows: C. albicans (23), C. parapsilosis (10), C. tropicalis (5), C. krusei (3), C. famata (2), C. glabrata (1), C. guilliermondii (1), C. kefyr (1). The antifungal susceptibilities were determined by BMD method described in Clinical and Laboratory Standards Institute M27-A3 document and E-test. Only two isolates (C. albicans and C. globrata) were found to be resistant to fluconazole with E-test but susceptible with BMD. The minimal inhibitory concentration (MIC) values of caspofungin were higher (MIC = 1-2 microg/ml) in C. parapsilosis compared to other Candida species using E-test. Only one C. albicans was resistant to voriconazole by E-test (MIC = 4 microg/ml), but it was susceptible by BMD (MIC = 0.08 microg/ml). Since definite resistance breakpoints do not yet exist for amphotericin B, MIC values were considered for amphotericin B and it was found that all strains had identical low MIC values (< 0.002-0.5). When E-test results were compared with the standard BMD results, MIC values were in agreement 80.4% for fluconazole, 84.7% for amphotericin B, 95.6% for voriconazole and 93.4% for caspofungin. These results indicated that the most frequently isolated Candida species among blood cultures was C. albicans, followed by C. parapsilosis and these isolates had low antifungal resistance rates. When voriconazol and caspofungin susceptibilities were considered, both E-test and BMD susceptibility results were in good aggreement in comparison to fluconazol and amphotericin B. E-test can be considered as a compatible method for the antifungal susceptibility testing of Candida species compared to standard broth microdilution method.

摘要

在过去几十年中,由于接受免疫抑制治疗的患者数量不断增加,严重真菌感染的发生率,尤其是侵袭性念珠菌感染呈上升趋势。这种情况最终导致对抗真菌药物的耐药性增加。本研究的目的是比较标准肉汤微量稀释法(BMD)和E-test法对我院血培养分离出的念珠菌属进行抗真菌药敏试验的结果,检测药物为氟康唑、伏立康唑、卡泊芬净和两性霉素B。2006年1月至2007年12月期间,通过BACTEC 9000(美国BD公司)从血培养中分离出并经传统技术和API 20C AUX(法国生物梅里埃公司)鉴定的46株念珠菌纳入本研究。分离株的鉴定结果如下:白色念珠菌(23株)、近平滑念珠菌(10株)、热带念珠菌(5株)、克柔念珠菌(3株)、法塔念珠菌(2株)、光滑念珠菌(1株)、季也蒙念珠菌(1株)、凯菲念珠菌(1株)。采用临床和实验室标准协会M27-A3文件中描述的BMD法和E-test法测定抗真菌药敏性。仅发现两株分离株(白色念珠菌和光滑念珠菌)E-test法检测对氟康唑耐药,但BMD法检测敏感。使用E-test法时,近平滑念珠菌中卡泊芬净的最低抑菌浓度(MIC)值较高(MIC = 1-2μg/ml),高于其他念珠菌属。E-test法检测仅1株白色念珠菌对伏立康唑耐药(MIC = 4μg/ml),但BMD法检测敏感(MIC = 0.08μg/ml)。由于两性霉素B尚无明确的耐药断点,因此对两性霉素B考虑MIC值,发现所有菌株的MIC值均较低(< 0.002-0.5)。将E-test结果与标准BMD结果进行比较时,氟康唑的MIC值一致性为80.4%,两性霉素B为84.7%,伏立康唑为95.6%,卡泊芬净为93.4%。这些结果表明,血培养中最常分离出的念珠菌属为白色念珠菌,其次是近平滑念珠菌,这些分离株的抗真菌耐药率较低。考虑伏立康唑和卡泊芬净的药敏情况时,与氟康唑和两性霉素B相比,E-test法和BMD法的药敏结果一致性良好。与标准肉汤微量稀释法相比,E-test法可被视为一种用于念珠菌属抗真菌药敏试验的兼容方法。

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