两种酵母样真菌药敏培养基检测氟康唑药敏试验的比较

目的比较丹麦ROSCO公司改良的Shadomy琼脂与M-H琼脂两种酵母真菌药敏培养基对氟康唑药敏试验结果的一致性。方法以美国临床实验室标准化委员会(NCCLS)M44-P推荐的酵母样真菌药敏培养基为参照,采用改良的Shadomy琼脂真菌药敏培养基检测氟康唑对常见临床分离的86株酵母样真菌的药敏试验。并以标准菌株作质量控制。结果两种培养基对氟康唑的药敏结果一致率为95.3%。经配对χ2检验,没有显著性差异(P>0.05)。结论选用配方不同的酵母样真菌药敏培养基适用于氟康唑药物纸片扩散法的测试,可以代替NCCLS M44-P酵母菌纸片扩散法在临床实验室使用。     

Rosco纸片扩散法检测酵母样真菌对氟康唑药敏试验的评价

目的　Rosco纸片扩散法在酵母样真菌药敏试验中的临床应用评价。方法　应用Rosco纸片扩散法和美国临床实验室标准化委员会 (NCCLS)M 2 7 A大量肉汤稀释法测定临床分离的 76株不同酵母样真菌菌株对氟康唑的药敏状况 ,并以 3株标准菌株作质量控制。结果　两种方法完全符合率是 90 8%未出现一种方法测得的敏感或耐药菌株 ,在用另一种方法检测中为耐药或敏感的严重错误。结论　Rosco纸片扩散法可以代替NCCLSM 2 7 A大量肉汤稀释法在临床推广使用。     

恶性肿瘤患者念珠菌临床分离株的药物敏感性分析

目的 以美国临床实验室标准化委员会（NCCLS） M27-A方案的微量肉汤稀释法为金标准,评价丹麦ROSCO公司的纸片扩散法在检测念珠菌耐药性方面的应用价值,为临床实验室寻找一种简便的念珠菌药敏试验方法.方法 分别采用丹麦ROSCO公司抗真菌药敏纸片和法国生物梅里埃公司ATBFUNGUR2念珠菌药敏板条来检测78株常见念珠菌对5-氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑等4种抗真菌药物的敏感性,以NCCLS的微量稀释法作为金标准,评价纸片扩散法的灵敏度、特异度、阳性预测值、阴性预测值.结果 纸片扩散法检测5-氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑的药敏结果,其Kappa值达到了0.89,未出现一种方法敏感而另一种方法耐药的严重错误现象;对78株念珠菌的药敏结果进行分析,5-氟胞嘧啶、两性霉素B敏感性高,分别为88.20％和89.17％,氟康唑和伊曲康唑敏感性较低,分别为56.34％和52.12％.白色念珠菌和热带念珠菌对4种抗真菌药物的敏感性较高,分别为90.95％、85.71％,而光滑念珠菌和克柔念珠菌的敏感性低,分别为67.50％、41.67％.结论 纸片扩散法与微量稀释法一致性高,在临床实验室可以替代微量稀释法进行念珠菌的药敏分析.我院念珠菌对两性霉素B的敏感性最高,对伊曲康唑的敏感性最低;抗真菌药物对白色念珠菌的抑菌率最高,对克柔念珠菌的抑菌率最低.     

两种药敏培养基在酵母菌体外药敏试验中的临床应用评价

目的比较两种药敏琼脂在真菌药敏试验中的临床应用效果。方法应用ROSCO抗真菌药纸片分别在RPMI 1640琼脂和葡萄糖美兰M—H（GMBMH）琼脂上的纸片扩散法（K—B法）及美国临床实验室标准化委员会（NCCLs）M27-A宏量肉汤稀释法测定192株临床分离的酵母样真菌对两性霉素、氟康唑、氟胞嘧啶、伊曲康唑、酮康唑的药敏情况。结果RPMI 1640和GMBMH对宏量肉汤稀释法的相关系数分别为0．93、0．82。未出现一种方法测得的敏感或耐药菌株在另一种方法中表现为耐药或敏感的严重错误。结论RPMI1640和GMB-MH对NCCI。SM27-A宏量肉汤稀释法测定结果均表现出较好相关性。但对NCCLs M27-A宏量肉汤稀释法的相关程度,RPMI 1640优于GMB-MH。RPMI 1640纸片扩散法可以替代NCCLs M27-A在临床推广使用。     

122株标本中分离的临床常见酵母样真菌的分类和对氟康唑的敏感性分析

酵母样真菌感染是临床上最常见的真菌感染之一,常见于免疫缺陷患者、长期使用内置导管进行治疗的患者以及内分泌失常的患者.随着抗真菌药物的广泛应用,酵母样真菌对常见抗真菌药物的敏感性逐渐降低,依据NCCLS推荐的纸片法评估酵母菌对氟康唑的敏感性,来帮助指导临床用药.……     

应用氟康唑对丹麦ROSCO公司与美国临床实验室标准化委员会酵母菌纸片扩散法药敏试验比较分析

目的评价丹麦ROSCO酵母菌纸片扩散法药敏试验及其判定标准在我院应用的可行性。方法以美国临床实验室标准化委员会（NCCLS）酵母菌纸片扩散法药敏试验为参照，采用丹麦ROSCO纸片扩散法药敏试验检测氟康唑对80株临床分离酵母菌的药敏试验。结果ROSCO纸片扩散法药敏试验选取基本判读标准，氟康唑药敏结果与NCCLS结果差异无统计学意义（P〉0．05）；而ROSCO选取严格判读标准，白念珠菌的氟康唑药敏结果与NCCLS结果存在差异，热带念珠菌结果差异无统计学意义（P〉0．05）。结论丹麦ROSCO酵母菌纸片扩散法药敏试验基本判读标准结果与美国NCCLS判读结果相符合，适合我院临床菌株的结果判读；而对于严重系统感染菌株所选取的严格判读标准，不适合白念珠菌的结果判读。     

酵母菌对氟康唑的敏感性及三种药敏试验方法的比较

目的 了解母菌对氟康唑的敏感性，并地３种抗真菌药敏试验方法进行比较。方法 利用纸片扩散法，微量稀释法及浓度梯度法（Ｅｔｅｓｔ法）测定９２株临床分离酵母菌对氟康唑的敏感性。结果 Ｅｔｅｓｔ法和微量稀释法所测酵母菌对氟康唑的总敏感率分别是７３．９％和８１．５％。纸片扩散法所测敏感菌，微量稀释法全部敏感，Ｅｔｅｓｔ法敏感率普９１％；纸片扩散法所测而药菌，其余两种方法检测多数仍为敏感。从不同菌种来看，白念     

2种药敏试验方法检测酵母菌对氟康唑敏感性的比较

目的研究纸片扩散法和真菌耐药性分析试剂盒(CANDIFAST)检测酵母菌对氟康唑敏感性结果的差异.方法同时应用美国临床实验室标准化委员会(NCCLS)认可的酵母菌纸片扩散法和CANDIFAST检测113株临床分离酵母菌对氟康唑的敏感性,并与临床资料进行比较.结果纸片扩散法和CANDIFAST所测酵母菌对氟康唑的总敏感率分别是93.8%(106/113)和54.0%(61/113).2种方法检测的结果差异有显著性(P<0.01),与临床治疗结果相比较,纸片扩散法的符合率平均高达99.5%,明显高于CANDIFAST(符合率平均63.8%).结论 NCCLS认可的酵母菌纸片扩散法的测定准确性优于CANDIFAST.     

检测常见酵母菌耐药性的纸片法和微量稀释法比较

目的用纸片法和微量稀释法检测痰液标本分离酵母菌的耐药性并比较两者符合率,试找出一种适用于临床实验室的简便方法。方法采用丹麦ROSCO公司抗真菌药敏纸片法检测痰液标本分离的86株常见酵母菌对5种抗真菌药物的敏感性,同时用法国生物梅里埃公司ATBFUNGUS2念珠菌药敏板条测定4种抗真菌药物的最小抑菌浓度,对结果进行分析。结果两性霉素B、氟胞嘧啶、氟康唑2种方法完全符合率分别为95．3％、95．3％和88．4％,未出现一种方法敏感而另一种方法耐药的严重错误现象;对86株痰液标本分离常见酵母菌的耐药性进行分析,两性霉素B、制霉菌素和氟胞嘧啶的敏感率高,分别为95．4％、98．8％、95．3％,伊曲康唑的耐药率最高（15．1％）。结论纸片法和微量稀释法相比有很好的一致性,且抗真菌药敏纸片种类多,便于随时根据临床所需增加单个药敏结果,易于在各临床实验室开展。酵母菌唑类抗真菌药物的耐药现象日益严重,应加强酵母菌的耐药性监测。     

酵母菌对氟康唑的敏感性及三种药敏试验方法的比较

目的了解酵母菌对氟康唑的敏感性，并对３种抗真菌药敏试验方法进行比较。方法利用纸片扩散法，微量稀释法及浓度梯度法（Ｅｔｅｓｔ法）测定９２株临床分离酵母菌对氟康唑的敏感性。结果Ｅｔｅｓｔ法和微量稀释法所测酵母菌对氟康唑的总敏感率分别是７３．９％和８１．５％。纸片扩散法所测敏感菌，微量稀释法全部敏感，Ｅｔｅｓｔ法敏感率为９１％；纸片扩散法所测耐药菌，其余两种方法检测多数仍为敏感。从不同菌种来看，白念珠菌、热带念珠菌、假热带念珠菌、葡萄牙念珠菌、白吉利丝孢酵母菌及酿酒酵母菌对氟康唑全部敏感；氟康唑对克柔念珠菌及光滑念珠菌的最小抑菌浓度（ＭＩＣ）值均较高；近平滑念珠菌、季也蒙念珠菌、异常汉逊酵母菌及新型隐球菌中的部分菌株对氟康唑敏感，部分菌株为剂量依赖性敏感，未发现耐药菌株。结论氟康唑对除克柔念珠菌及光滑念珠菌的ＭＩＣ值较高外，其余酵母菌对氟康唑敏感性好。纸片扩散法虽简便，但仅可用于初筛敏感菌，对抑菌环直径≤１４ｍｍ者应进一步用微量稀释法或Ｅｔｅｓｔ法测定其ＭＩＣ值     

检测常见酵母菌耐药性的纸片法和微量稀释法比较

目的用纸片法和微量稀释法检测痰液标本分离酵母菌的耐药性并比较两者符合率,试找出一种适用于临床实验室的简便方法。方法采用丹麦ROSCO公司抗真菌药敏纸片法检测痰液标本分离的86株常见酵母菌对5种抗真菌药物的敏感性,同时用法国生物梅里埃公司ATB FUNGUS 2念珠菌药敏板条测定4种抗真菌药物的最小抑菌浓度,对结果进行分析。结果两性霉素B、氟胞嘧啶、氟康唑2种方法完全符合率分别为95.3%、95.3%和88.4%,未出现一种方法敏感而另一种方法耐药的严重错误现象;对86株痰液标本分离常见酵母菌的耐药性进行分析,两性霉素B、制霉菌素和氟胞嘧啶的敏感率高,分别为95.4%、98.8%、95.3%,伊曲康唑的耐药率最高(15.1%)。结论纸片法和微量稀释法相比有很好的一致性,且抗真菌药敏纸片种类多,便于随时根据临床所需增加单个药敏结果,易于在各临床实验室开展。酵母菌唑类抗真菌药物的耐药现象日益严重,应加强酵母菌的耐药性监测。     

Comparison of Etest and a tablet diffusion test with the NCCLS broth microdilution method for fluconazole and amphotericin B susceptibility testing of Candida isolates

Three methods were compared for the susceptibility testing of yeast isolates to fluconazole and amphotericin B: two fagar diffusion methods (Etest and a tablet diffusion test) and the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method. Given as MIC(50)s (range), fluconazole endpoints were: for the 24 h broth microdilution test, 0.25 mg/L (0.06-32 mg/L); for the Etest, 0.38 mg/L (0.064-24 mg/L); and for the NCCLS broth microdilution test, 2 mg/L (0.06->or=64 mg/L). With breakpoints of <3 mg/L for susceptible and >16 mg/L for resistant, the Etest and the 24 h microdilution test classified the isolates in agreement with the classification obtained by the NCCLS method. Results obtained by Etest were in closer NCCLS method than those obtained with the tablet test. Amphotericin B endpoints were lower for the 24 h microdilution and Etests than MICs obtained by the NCCLS broth microdilution method. Reproducibility was high for all tests; however, disadvantages of both diffusion tests were microcolonies in the inhibition zone and dependence on stringent standardization of inoculum.     

Correlation between E-test,disk diffusion,and microdilution methods for antifungal susceptibility testing of fluconazole and voriconazole

The activities of fluconazole and voriconazole against isolates of Candida spp. (n = 400) were tested by the E-test, disk diffusion, and the National Committee for Clinical Laboratory Standards (NCCLS) M27-A2 broth microdilution-based reference methods. More than 96% of isolates found to be susceptible to fluconazole by the reference method were identified as susceptible by the agar-based methods. Lesser degrees of correlation with the reference method were seen for isolates identified as resistant by the agar-based methods. Interpretive categories are not available for voriconazole, but results qualitatively similar to those for fluconazole were seen. The agar-based E-test and disk diffusion methods are reliable alternatives to the NCCLS M27-A2 reference microdilution method for isolates that test susceptible to fluconazole.     

氟康唑体外抗菌活性及五种体外敏感试验方法的比较

目的 对比研究五种酵母菌体外药敏试验方法在测定氟康唑对酵母菌的体外活性检测上的可靠性及实用性。方法 分别应用美国临床实验室标准化委员会（NCCLS）M27-A常量肉汤稀释法、微量肉汤稀释法，NCCLS及ROSCO纸片扩散法、浓度梯度法测定氟康唑对155株临床分离的酵母菌及4株质控菌株的体外活性。采用WHONET-5软件及SPSS软件对结果进行分析，比较各种药敏试验方法与M27-A常量肉汤稀释法的相关性。结果 4种体外药敏试验方法所得结果分别同M27-A常量肉汤稀释法进行比较，浓度梯度法一致率为83．9％，NCCLSM44-P纸片扩散法一致率83．1％，ROSCO纸片扩散法一致率为78．1％，微量肉汤稀释法一致率为93．5％。结论 5种酵母菌体外药敏试验方法在检测氟康唑对临床常见酵母菌的体外活性检测上存在一定的差异，本次试验结果表明，微量肉汤稀释法与NCCLSM27-A常量肉汤稀释法的一致性最佳，其方法结果准确可靠、重复件好．活用于常规下作。     

Clinical evaluation of a dried commercially-prepared microdilution panel for antifungal susceptibility testing

A commercially-prepared dried broth microdilution panel (Sensititre) was compared with a reference microdilution method for antifungal susceptibility testing of two reference yeast strains and 98 clinical isolates of Candida spp. The antifungal agents tested include 5-fluorocytosine (5FC), fluconazole, itraconazole, and D0870. Microdilution testing was performed according to National Committee for Clinical Laboratory Standards (NCCLS) recommendations. Minimum inhibitory concentration (MIC) endpoints were read visually after 48 hours of incubation and were assessed independently for each microdilution panel. The MICs for the reference strains were within published control limits for both reference and Sensititre microdilution panels. Discrepancies among MIC endpoints of no more than two dilutions (two wells) were used to calculate the percent agreement. An acceptable level of agreement between Sensititre and reference panels was observed for all antifungal agents when tested against the 98 clinical isolates. Agreement ranged from 83% for itraconazole to 93% for 5FC. The Sensititre dried microdilution panel appears to be a viable alternative to inhouse prepared microdilution panels and to the NCCLS macrodilution reference method.     

Protective effect of trovafloxacin, ciprofloxacin and ampicillin against Streptococcus pneumoniae in a murine sepsis model

The Etest strip is a promising tool of broad application in clinical microbiology. The method provides MIC readings and is easier to perform than broth microdilution. We carried out a study to compare the MICs of fluconazole and itraconazole obtained by the Etest with those obtained by broth microdilution, performed according to the guidelines of the NCCLS document M27-A, with 402 clinical isolates (360 Candida albicans, 17 Candida tropicalis, nine Candida krusei, nine Candida glabrata and seven Candida parapsilosis) and seven control isolates. The agreement between MICs by the two methods (at +/- 2 dilutions) was 74.5% for fluconazole and 61.4% for itraconazole. These results suggest that further development is necessary to standardize the medium and incubation conditions before introduction of the Etest as a routine method in the clinical microbiology laboratory for fluconazole and itraconazole susceptibility testing.     

Flow cytometry antifungal susceptibility testing of pathogenic yeasts other than Candida albicans and comparison with the NCCLS broth microdilution test

Candida species other than Candida albicans frequently cause nosocomial infections in immunocompromised patients. Some of these pathogens have either variable susceptibility patterns or intrinsic resistance against common azoles. The availability of a rapid and reproducible susceptibility-testing method is likely to help in the selection of an appropriate regimen for therapy. A flow cytometry (FC) method was used in the present study for susceptibility testing of Candida glabrata, Candida guilliermondii, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans based on accumulation of the DNA binding dye propidium iodide (PI). The results were compared with MIC results obtained for amphotericin B and fluconazole using the NCCLS broth microdilution method (M27-A). For FC, the yeast inoculum was prepared spectrophotometrically, the drugs were diluted in either RPMI 1640 or yeast nitrogen base containing 1% dextrose, and yeast samples and drug dilutions were incubated with amphotericin B and fluconazole, respectively, for 4 to 6 h. Sodium deoxycholate and PI were added at the end of incubation, and fluorescence was measured with a FACScan flow cytometer (Becton Dickinson). The lowest drug concentration that showed a 50% increase in mean channel fluorescence compared to that of the growth control was designated the MIC. All tests were repeated once. The MICs obtained by FC for all yeast isolates except C. lusitaniae were in very good agreement (within 1 dilution) of the results of the NCCLS broth microdilution method. Paired t test values were not statistically significant (P = 0.377 for amphotericin B; P = 0.383 for fluconazole). Exceptionally, C. lusitaniae isolates showed higher MICs (2 dilutions or more) than in the corresponding NCCLS broth microdilution method for amphotericin B. Overall, FC antifungal susceptibility testing provided rapid, reproducible results that were statistically comparable to those obtained with the NCCLS method.