临床分离真菌耐药性分析

目的监测分析临床分离真菌对抗菌药物的耐药现状,以加强抗真菌药物的合理应用。方法对我院2007年1月至2007年6月分离鉴定出的479株真菌用目前常用的5种抗真菌药物进行药物敏感性试验和分析。结果479株真菌中念珠菌和其他真菌分别占98.3%和1.7%,其中前3位依次是白念珠菌（69.5%）、光滑念珠菌（16.9%）和热带念珠菌（8.8%）。在5种抗真菌药物中,耐药率由高到低依次为伊曲康唑（12.7%）、氟康唑（9.8%）、伏立康唑（7.5%）、氟胞嘧啶（1.8%）和两性霉素B（1.0%）。结论临床分离的真菌对当前常用抗真菌药物的耐药菌株增多,应引起重视。     

肿瘤患者临床分离真菌的分布及耐药性分析

目的监测分析临床分离真菌的分布以及抗真菌药物的耐药现状,加强抗真菌药物的合理应用。方法对2009年1～12月分离鉴定出的210株真菌用目前常用的5种抗真菌药物进行药敏试验和分析。结果 210株真菌中念珠菌和其他真菌分别占96.7%和3.3%,其中居前4位的念珠菌依次是白念珠菌(70.5%)、光滑念珠菌(12.9%)、热带念珠菌(8.2%)和克柔念珠菌(3.8%)。呼吸道标本检出率最高,肺癌患者检出率最高。两性霉素B耐药率最低,氟康唑和伊曲康唑对光滑念珠菌和克柔念珠菌耐药率高。结论肿瘤患者临床分离的耐氟康唑光滑念珠菌和克柔念珠菌有升高趋势,微生物室应将念珠菌鉴定到种并对其进行个体化药敏试验,加强耐药性监测。     

281株临床分离念珠菌的药敏结果分析

目的 对临床送检标本分离鉴定出的念珠菌进行细菌构成及药敏结果分析,促进临床合理使用抗真菌药物.方法 对2009年1月至2010年12月临床送检标本进行 5种常用抗真菌药(5-氟胞嘧啶、氟康唑、伊曲康唑、伏立康唑、两性霉素B)的体外敏感性检测及耐药分析.结果 281株念珠菌中白色念珠菌184株,占65.5%,其次是光滑念珠菌50株(17.8%),热带念珠菌19株(6.8%).结论 该院患者念珠菌感染以白色念珠菌为主,药敏结果显示两性霉素B、5-氟胞嘧啶、伏立康唑、氟康唑、伊曲康唑敏感性高,对送检标本及时进行真菌培养和药敏试验,可及时、合理地使用抗真菌药物,减少多重耐药和深部真菌感染的发生.     

2019—2021年某教学医院无菌体液样本分离真菌临床分布及耐药性变迁

目的 分析合肥市第一人民医院临床送检无菌体液样本分离真菌的临床分布及其耐药性变迁,为临床合理使用抗真菌药物提供参考。方法 收集2019—2021年合肥市第一人民医院无菌体液样本非重复真菌临床分离株200株。采用显色培养基分离、培养后,经基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)进行菌种鉴定,并用真菌药物敏感性试验板条分析其耐药性。比较3年中200株真菌临床分布及其对常用抗真菌药物的耐药性。结果 2019、2020、2021年分离自无菌体液样本的真菌分别占分离自所有临床样本的真菌的10.7%、26.7%、17.0%;主要分离自引流液、腹腔积液、支气管灌洗液;科室分布主要为重症监护病房、外科、呼吸科;主要分离自60岁以上老年人群。菌种类型居前3位的依次为白念珠菌、热带念珠菌和光滑念珠菌。2019、2020、2021年,白念珠菌对伊曲康唑的耐药率分别为10.5%、8.9%、10.6%,对氟康唑的耐药率分别为5.2%、2.9%、6.1%;光滑念珠菌对伊曲康唑的耐药率分别为25.0%、33.3%、28.6%,对氟康唑的耐药率分别为0、16.7%和0;热带念珠菌对伊曲康唑的耐药率...     

临床分离念珠菌对5种常用抗真菌药物的体外敏感试验结果分析

目的了解临床分离的念珠菌对氟康唑、两性霉素B、氟胞嘧啶、伊曲康唑及酮康唑体外敏感性.方法采用Sensititre YeastOne试验板以微量稀释法测定上述5种抗真菌药物对临床分离的108株念珠菌最低抑菌浓度(MIC).结果 108株念珠菌中达到氟康唑、伊曲康唑、氟胞嘧啶耐药标准的分别有8株(7.4%)、15株(13.9%)、2株(1.9%),念珠菌属MIC值分布种间差异较大.白色念珠菌对5种药物的MIC90值最低,60株白色念珠菌中仅2株耐氟康唑,3株耐伊曲康唑,对氟胞嘧啶无耐药株;光滑念珠菌对氟康唑、伊曲康唑、酮康唑的MIC值分布呈高值,10株光滑念珠菌中4株耐氟康唑,3株剂量依赖性敏感,7株耐伊曲康唑,且吡咯类之间有交叉耐药.其他菌株,除季也蒙念珠菌对伊曲康唑有一定的耐药(2/6)外,对5种抗真菌药物的MIC分布均较低.结论不同念珠菌对常用抗真菌药物敏感性存在差异,准确分离鉴定和药敏试验,对于指导临床合理选药有重要意义.     

临床分离念珠菌对5种常用抗真菌药物的体外敏感试验结果分析

目的　了解临床分离的念珠菌对氟康唑、两性霉素B、氟胞嘧啶、伊曲康唑及酮康唑体外敏感性。方法　采用SensititreYeastOne试验板以微量稀释法测定上述 5种抗真菌药物对临床分离的 10 8株念珠菌最低抑菌浓度 (MIC)。结果　 10 8株念珠菌中达到氟康唑、伊曲康唑、氟胞嘧啶耐药标准的分别有 8株 (7.4%)、15株(13.9%)、2株 (1.9%) ,念珠菌属MIC值分布种间差异较大。白色念珠菌对 5种药物的MIC90 值最低 ,6 0株白色念珠菌中仅 2株耐氟康唑 ,3株耐伊曲康唑 ,对氟胞嘧啶无耐药株 ;光滑念珠菌对氟康唑、伊曲康唑、酮康唑的MIC值分布呈高值 ,10株光滑念珠菌中 4株耐氟康唑 ,3株剂量依赖性敏感 ,7株耐伊曲康唑 ,且吡咯类之间有交叉耐药。其他菌株 ,除季也蒙念珠菌对伊曲康唑有一定的耐药 (2 /6 )外 ,对 5种抗真菌药物的MIC分布均较低。结论　不同念珠菌对常用抗真菌药物敏感性存在差异 ,准确分离鉴定和药敏试验 ,对于指导临床合理选药有重要意义。     

念珠菌420株临床分布及Rosco纸片法药敏结果分析

目的了解本院念珠菌的分布及对常用抗真菌药物的耐药情况,以更好地配合临床医生合理用药。方法收集2008-01-2008-12临床分离标本420株,应用Rosco纸片扩散法检测7种抗真菌药物对临床常见酵母样真菌的耐药性。结果 420株念珠菌菌中,白色念珠菌、热带念珠菌、光滑念珠菌、克柔念珠菌和近平滑念珠菌分别占61.9%、17.0%、14.5%、3.5%和2.9%;分离株对两性霉素B、伏立康唑、伊曲康唑、氟康唑、酮康唑、5-氟胞嘧啶和益康唑的敏感率依次为100%、100%、97.8%、94.3%、89.5%、88.1%和84.5%。结论临床微生物实验室要加强对念珠菌的分离培养和药敏试验,指导临床合理用药,有效控制和减少真菌感染。     

118株酵母样真菌药敏结果分析

目的分析酵母样真菌对目前常用的5种抗真菌药物的敏感性,为临床合理选用抗真菌药物提供依据。方法对云南省第一人民医院2009年1～6月分离的118株酵母样真菌的药敏试验结果进行回顾性分析。结果酵母样真菌感染以白色念珠菌为主,酵母样真菌对5-氟胞嘧啶、两性霉素B敏感率较高,在98%以上,对唑类药物敏感性稍低,为70%～80%。结论酵母样真菌感染以白色念珠菌、热带念珠菌和光滑念珠菌较为常见,而且耐药菌株的检出率呈上升趋势,临床上应慎用抗菌药物,减少耐药菌株的产生。     

宫颈癌患者腔内放疗合并局部真菌阴道感染分析

目的：了解我院宫颈癌患者腔内放疗合并局部真菌阴道感染菌种分布及其耐药特征,以便指导临床合理用药。方法对本院2009年1月-2014年12月妇科宫颈癌患者实施腔内后装放疗期间出现局部感染的病例695例,采集阴道分泌物进行真菌培养和药敏试验并进行统计分析。结果共分离获得真菌89株,真菌分离率为12.8％。以白念珠菌为主(62株,69.7％),其次是光滑念珠菌(12株,13.5％)及热带念珠菌(9株,10.1％)。分离的真菌对5-氟胞嘧啶、两性霉素B、制霉菌素耐药率较低,而对咪康唑、益康唑、氟康唑、伊曲康唑均有不同程度的耐药。结论宫颈癌患者腔内放疗后阴道分泌物真菌分离率较高,应引起临床医务人员高度重视;主要分离真菌对临床常用抗真菌药物有一定的耐药性,临床应依据药敏试验合理使用抗真菌药物。     

上海地区临床分离念珠菌的耐药性监测

目的了解上海地区临床分离念珠菌的菌株分布以及对常用抗真菌药物的敏感性和耐药性。方法对上海地区27所医院临床分离念珠菌采用微量液基稀释法按统一方案进行抗真菌药物敏感性试验。按CLSI M60(2017年)判断结果。结果收集2017年8月1日-2018年4月30日上述医院临床分离念珠菌共354株,其中白念珠菌占41.81%(148/354),近平滑念珠菌占23.73%(84/354),热带念珠菌占15.54%(55/354),光滑念珠菌占9.32%(33/354),其他念珠菌占9.60%(34/354)。白念珠菌、近平滑念珠菌、光滑念珠菌耐药率与以往报道比较偏低或基本持平,无增长趋势,主要是对唑类抗真菌药物具有一定的耐药性。白念珠菌对氟康唑耐药率为1.45%、剂量依赖性敏感(SDD)率为4.35%,对伏立康唑耐药率和中介率均为2.17%;近平滑念珠菌对氟康唑呈现1.20%的SDD率,对伏立康唑呈现1.20%的中介率;光滑念珠菌对氟康唑表现出6.06%的耐药率和93.94%的SDD率;热带念珠菌耐药率呈增长趋势,对氟康唑表现出41.82%的耐药率和12.73%的SDD率,对伏立康唑表现出40.00%的耐药率和12.73%的中介率。白念珠菌、近平滑念珠菌、热带念珠菌对棘白菌素类药物尚未出现耐药菌株,仅1株光滑念珠菌对卡泊芬净耐药。结论上海地区临床念珠菌的分离率及菌株分布与国内外报道基本一致,临床分离菌株对常用抗菌药物的耐药率基本稳定,热带念珠菌对唑类抗真菌药物的耐药率呈上升趋势,应该引起临床医师重视。同时应进一步加强医院感染防控措施,继续做好真菌耐药性监测工作。     

深部酵母菌感染的病原学特征及其药物敏感性

目的　了解深部酵母菌感染的病原学特征及其对常用抗真菌药物的敏感性。方法　使用常规方法或用商品试剂盒包括API 2 0CAUX和VITEKYBC生化卡鉴定菌种 ;使用E test检测部分菌株对氟康唑、酮康唑、伊曲康唑和两性霉素B等 4种抗真菌药的敏感性。结果　 1998年 5月～ 2 0 0 2年 5月 4年间从深部标本中共分离出酵母菌 30 7株 ,其中念珠菌占 94 .79% ;检出率居前 4位的菌种为 :白色念珠菌 (4 7.88% )、热带念珠菌(2 6 .72 % )、光滑念珠菌 (8.4 7% )、近平滑念珠菌 (7.4 9% )。念珠菌对氟康唑和伊曲康唑的耐药率分别为 12 .8%和 13.9%。结论　不同标本中各菌种所占比例有所不同 ,不同菌种对不同药物的敏感性也不同。     

Reliability of the WIDERYST susceptibility testing system for detection of in vitro antifungal resistance in yeasts

This study evaluated the WIDERYST system, a commercially available computer-assisted image-processing device for the antifungal susceptibility testing of yeasts. A collection of 90 clinical isolates selected to represent ranges of susceptibilities in vitro as broad as possible was tested. An evaluation compared the results obtained by the new system with those achieved by both the Clinical and Laboratory Standards Institute (CLSI) microdilution reference procedure and the antifungal susceptibility standard of the European Committee for Antimicrobial Susceptibility Testing (EUCAST). Overall, the agreement and the correlation index between results obtained by the EUCAST method and the WIDERYST system were 89% and 0.84 (P < 0.01), respectively, and agreement and correlation index between data obtained by the CLSI procedure and the WIDERYST system were 90% and 0.86 (P < 0.01), respectively. The system was able to detect amphotericin B-resistant isolates. All Candida sp. isolates with resistance in vitro to azole agents were detected as well. The system misclassified some isolates belonging to the slowly growing genera Dipodascus and Pichia. A total of 2.7% very major errors were detected for fluconazole. The WIDERYST system is an alternative to reference procedures for antifungal susceptibility testing of clinical isolates of yeasts, particularly for Candida and Cryptococcus species.     

Antifungal susceptibility testing of yeasts: evaluation of technical variables for test automation.

The technical parameters for antifungal susceptibility testing with Candida species were reexamined to determine the optimal conditions for testing with semiautomated preparations of broth microdilution cultures, automated spectrophotometric readings of the cultures, and dose-response and endpoint determinations by means of a computer spreadsheet. Tests were based on proposed standard method M27P of the National Committee for Clinical Laboratory Standards for antifungal agents. RPMI 1640 broth with extra glucose to a final concentration of 2% gave higher and more reproducible drug-free control readings without affecting susceptibility endpoint readings. An inoculum of 8 x 10(4) yeasts per ml prepared from a carbon-limiting broth culture without further standardization was found to give optimal control readings after 48 h of incubation at 37 degrees C. For flucytosine, fluconazole, itraconazole, and ketoconazole, endpoints based on 50% growth inhibition (50% inhibitory concentration) gave the minimum variation with inoculum size and the fewest endpoint differences with RPMI 1640 medium obtained from two different suppliers. The 50% inhibitory concentration was also the optimal endpoint for fluconazole and ketoconazole susceptibilities in comparison with broth macrodilution MICs determined by the method of the National Committee for Clinical Laboratory Standards. Intralaboratory reproducibility was determined by retrospective analysis of replicate results for isolates retested at random over a 2-year period. This approach showed less favorable reproducibility than has been reported from purpose-designed, prospective antifungal susceptibility studies, but it may better reflect real-life test reproducibility. Susceptibility data for 616 clinical isolates of yeasts, representing 16 Candida and Saccharomyces spp., confirmed the tendency of Candida lusitaniae isolates to show relatively low susceptibilities to amphotericin B, the tendency of Candida krusei isolates to show low flucytosine and fluconazole susceptibilities, and the presence of some isolates in the species Candida albicans, Candida glabrata, and Candida tropicalis with low susceptibilities to azole derivative antifungal agents. The study demonstrates the value of automation and standardization in all stages of yeast susceptibility testing, from plate preparation to data analysis.     

E试验测定78株念珠菌属真菌对4种抗真菌药物的敏感性

目的 检测4种抗真菌药物对念珠菌属真菌的最低抑菌浓度（MIC）并分析其耐药性。方法 用E试验检测两性霉素B（AP）、酮康唑（KE）、氟康唑（FL）、伊曲康唑（IT）对78株临床分离的念珠菌属真菌的MIC值。结果 78株念珠菌属真菌对AP、KE、FL、IT的总敏感率分别是89．7％、92．3％、85．9％、85．9％；白念珠菌对AP、KE、FL和IT的敏感率分别是96．9％、96．9％、100％和87．5％。结论 4种药物对白念珠菌的体外抗菌活性均较好，但对非白念珠菌的抗菌活性则较前者为差。     

Head-to-head comparison of the activities of currently available antifungal agents against 3,378 Spanish clinical isolates of yeasts and filamentous fungi

We have compared the activities of posaconazole and other currently available antifungal agents against a collection of 3,378 clinical isolates of yeasts and filamentous fungi. A total of 1,997 clinical isolates of Candida spp., 359 of other yeast species, 697 strains of Aspergillus spp., and 325 nondermatophyte non-Aspergillus spp. were included. The average geometric means of the MICs of agents that were tested against Candida spp. were 0.23 microg/ml for amphotericin B, 0.29 microg/ml for flucytosine, 0.97 microg/ml for fluconazole, 0.07 microg/ml for itraconazole, 0.04 microg/ml for voriconazole, 0.15 microg/ml for caspofungin, and 0.03 microg/ml for posaconazole. Voriconazole and posaconazole were active in vitro against the majority of isolates, with resistance to fluconazole and itraconazole, and against Cryptococcus neoformans and other Basidiomycota yeasts. Posaconazole was the most active of antifungal agents tested against Aspergillus spp., with an average geometric mean of 0.10 microg/ml. It was active against Paecilomyces spp., Penicillium spp., Scedosporium apiospermum, and some black fungi, such as Alternaria spp. Multiresistant filamentous fungi, such as Scedosporium prolificans, Scopulariopsis brevicaulis, and Fusarium solani, were also resistant to voriconazole, caspofungin, and posaconazole. Amphotericin B and posaconazole were found to be active against most of the Mucorales strains tested. Posaconazole and currently available antifungal agents exhibit a potent activity in vitro against the majority of pathogenic fungal species.     

In vitro susceptibility of Candida dubliniensis to current and new antifungal agents

BACKGROUND: Candida dubliniensis is a recently described Candida species closely related to Candida albicans, which has been associated with oral candidiasis in HIV-infected patients. Fluconazole-resistant strains of C. dubliniensis are easily obtained in vitro and this fact could be a complication if this resistance develops during treatment with this drug. METHODS: In the present study, the in vitro antifungal susceptibilities of 36 C. dubliniensis clinical isolates and culture strains to current and new antifungal agents, such as amphotericin B (AMB), amphotericin B lipid complex (ABLC), amphotericin B colloidal dispersion (ABCD), 5-fluorocytosine (5FC), fluconazole (FLC), itraconazole (ITC), ketoconazole (KTC), liposomal amphoteri- cin B (LAMB), liposomal nystatin (LNYT), LY303366 (LY), SCH56592 (SCH), and voriconazole (VRC), were determined according to the National Committee for Clinical Laboratory Standards M27-A broth microdilution method for yeasts. RESULTS: Most isolates of C. dubliniensis were susceptible to both new and current antifungal drugs, with 75.9% isolates susceptible to KTC, 86.2% to FLC and to ITC, and approximately 100% to the other antifungal agents tested. The cross-resistance phenotypes are detailed. Four isolates were resistant (MIC > or =64 microg/ml) to FLC. These 4 isolates were also resistant to KTC, and 3 of them were also resistant to ITC (MIC > or =1 microg/ml for both agents). However, these isolates were highly susceptible to 5FC and all polyene formulations (AMB, ABLC, ABCD, LAMB, and LNYT), triazole (SCH and VRC) and echinocandin (LY) antifungal agents. CONCLUSION: The new liposomal and lipidic formulations of AMB, LNYT, and the new triazoles and echinocandins may provide new alternatives to FLC for the treatment of infections by C. dubliniensis.     

Isolation of Candida species on media with and without added fluconazole reveals high variability in relative growth susceptibility phenotypes.

Mouthwashes from human immunodeficiency virus-positive individuals were sampled for yeasts by direct plating on a differential agar medium with and without added fluconazole and via enrichment broths with and without added fluconazole. The colonies of the yeasts isolated were tested for relative growth in the presence of single concentrations of itraconazole and fluconazole. Among 258 culture plates containing yeasts obtained via different isolation routes from 86 yeast-positive samples, 33 (12.7%) of the plates showed unexpectedly high colony-to-colony variation in relative growth. Intercolony variation was seen in 41 (47.7%) of the 86 isolates when relative growth data were analyzed for all colonies of an isolate tested, regardless of the medium used for isolation. The prevalence of relative growth variability with the azoles was highest for Candida glabrata (100% of 13 isolates), followed by Candida krusei (60% of 5 isolates) and Candida albicans (40% of 53 isolates), and the visual patterns of variability seen in scatter plots of the data showed species specificity. Relative growth phenotypes generally tended to be stable for each yeast colony in subcultures, whether or not the medium used for subculture contained antifungal agents. DNA fingerprinting of stable and variable C. albicans isolates showed changes in band patterns detected with the probe Ca3, suggesting that the variability may have resulted from selection of different subtypes of the yeasts during the isolation procedure. These findings suggest that the yeasts isolated from single clinical samples were often not clonal in nature. The relative growth test revealed colony variability more readily than conventional susceptibility testing.     

In vitro susceptibility of Candida species to five antifungal agents in a German university hospital assessed by the reference broth microdilution method and Etest

OBJECTIVES: The aim of this study is to evaluate the susceptibilities of Candida spp. to the common antifungal agents in a German university hospital. Since quick results of in vitro testing are desirable, Etest and the CLSI broth microdilution (BMD) method (reference method) were compared, focusing on the validity of early readings. METHODS: A total of 512 Candida spp. isolates, including 174 from primarily sterile sites, were collected in the clinical routine. The yeasts were differentiated by CHROMagar and verified by API 20C AUX if necessary. In vitro susceptibilities to amphotericin B, flucytosine, fluconazole, voriconazole and caspofungin were determined using the BMD method described in the CLSI (formerly NCCLS) M27-A2 document and Etest. MICs were noted after 24 and 48 h of incubation. RESULTS: The most frequently isolated species was Candida albicans. Among the non-albicans species, Candida glabrata was the most prevalent, followed by Candida tropicalis, Candida parapsilosis and Candida krusei. MICs (mg/L) at which 90% of the strains were inhibited were 1 for amphotericin B, 32 for flucytosine, 8 for fluconazole, 0.25 for voriconazole and 1 for caspofungin. Susceptibility to fluconazole was 85.0% for C. glabrata and 5.3% for C. krusei, almost all other isolates were susceptible in over 90% except very rare species. The 48 h MIC values of Etest and BMD were in agreement (no more than 2 log(2) dilutions) in 88.7% to 98.1% with categorical agreement rates of 91.6% to 98.2%, depending on the antifungal agent. Comparison of the 24 h MICs of both BMD and Etest with the 48 h MICs of the reference method showed categorical agreement in 94.9% to 99.2%. For caspofungin, however, a comparison of the categorical agreement was not possible due to the lack of interpretive breakpoints. The order of frequency and the resistance patterns of the isolates from primarily sterile sites and those of isolates from non-sterile sites did not differ. CONCLUSIONS: No alarming resistances against the agents tested were found; however, owing to the relatively high frequency of C. glabrata with elevated fluconazole MICs, this species and, to a certain extent, C. krusei must be taken into consideration when choosing antifungal agents for calculated therapy. Etest is a reliable method for the susceptibility testing of Candida spp. and the 24 h readings of both Etest and BMD can serve as helpful preliminary results in most cases.