E-test法检测申克氏孢子丝菌对抗真菌药物敏感性的研究

为探讨申克氏孢子丝菌的体外抗真菌药敏试验方法的重复性及准确性,观察申克氏孢子丝菌对两种抗真菌药的敏感性,我们采用美国临床实验室标准委员会（NCCLS）颁布的M27-P方案中的E-test法检测申克氏孢子丝菌分别在菌丝相和酵母相时对伊曲康唑、氟康唑的MIC值。结果：25℃菌丝相时氟康唑、伊曲康唑的MIC值范围分别为：2～256μg/mL、0.012～8.0μg/mL。37℃酵母相时氟康唑、伊曲康唑的MIC值范围分别为：4～256μg/mL、0.125～32μg/mL。申克氏孢子丝菌菌丝相的MIC值较低,酵母相的MIC值较高。申克氏孢子丝菌25℃菌丝相和37℃酵母相时对伊曲康唑、氟康唑的敏感性存在差异。对伊曲康唑的敏感性高于氟康唑。     

61株甲源性近平滑念珠菌药敏试验报告

根据美国国家临床实验室标准化委员会M27-A微量稀释法对近平滑念珠菌作了伊曲康唑、特比萘芬和氟康唑3种抗真菌药物敏感性测定。结果:伊曲康唑、特比萘芬、氟康唑对61株近平滑念珠菌的平均MIC值分别为:3.20μg/mL、0.42μg/mL、2.33μg/mL。体外特比萘芬对近平滑念珠菌敏感性高于伊曲康唑和氟康唑。     

E-test法和Rosco纸片法检测申克孢子丝菌对伊曲康唑和氟康唑的敏感性试验

目的探讨申克孢子丝菌的体外抗真菌药敏试验E-test法和Rosco纸片法的可比性,观察申克孢子丝菌对伊曲康唑和氟康唑的敏感性。方法采用临床实验室标准化协会(CLSI)颁布的M27-P方案中的E-test法检测和Rosco纸片法检测申克孢子丝菌在菌丝相时对伊曲康唑、氟康唑的MIC值。结果采用E-test法和Rosco纸片法检测20株临床分离株25℃菌丝相时,伊曲康唑的MIC值范围分别为:0.012～2.0μg/mL,14～29mm,氟康唑的MIC值范围分别为:256μg/mL,0mm。结论体外抗真菌药敏试验E-test法和Rosco纸片法有较好的一致性和重复性。申克孢子丝菌25℃菌丝相时对伊曲康唑、氟康唑的敏感性存在差异,对伊曲康唑的敏感性高于氟康唑。     

氟康唑治疗孢子丝菌病的临床疗效及体外抗真菌药敏试验研究

目的观察氟康唑对孢子丝菌病的临床疗效,检测孢子丝菌对氟康唑、伊曲康唑的体外敏感性,探讨孢子丝菌病治疗的新方法。方法对60例孢子丝菌病患者给予氟康唑片口服治疗[成人首次100mg,以后50mg,儿童3～5mg/(kg.d)],30例患者给予10%碘化钾溶液10mL口服治疗,并采用E-test法检测其中10株孢子丝菌对氟康唑和伊曲康唑的最小抑菌浓度(MIC)。结果氟康唑组有效率为95.00%,碘化钾组为86.67%。采用E-test法检测10株孢子丝菌对氟康唑和伊曲康唑的MIC值,范围分别为>256μg/mL,0.012～8.0μg/mL。结论氟康唑对孢子丝菌病有较好的临床疗效,但申克氏孢子丝菌对对氟康唑的体外敏感性较低。E-test法可以作为一种孢子丝菌的体外抗真菌药敏试验方法。     

51株甲真菌病白念珠菌体外药物敏感性试验

参照美国国家临床实验室标准化委员会M27-A棋盘微量稀释法,检测了分离自2005年1月至2007年10月,于北京大学第三医院第二门诊部就诊的甲真菌病患者的51株白念珠菌对伊曲康唑、特比萘芬和氟康唑3种抗真菌药物的体外敏感性.伊曲康唑、特比萘芬、氟康唑对51株甲源性白念珠菌的平均最小抑菌浓度(MIC)分别为:0.23 μg/mL、3.84 μg/mL、6.52 μg/mL.伊曲康唑在体外对白念珠菌敏感性高于特比萘芬和氟康唑.     

申克孢子丝菌双相体外药物敏感试验研究

目的：研究双相真菌申克孢子丝菌分别在菌丝相和酵母相两种形态下对常用抗真菌药物的敏感性。方法：分别采用Roseo纸片扩散法及微量液基稀释法对临床分离的4株申克孢子丝菌的两种形态（菌丝相及酵母相）进行体外药敏试验。测定的抗真菌药物为：两性霉素B、伊曲康唑、氟康唑、特比萘芬。结果：除AMB二者一致外，其余各抗真菌药的MIC，菌丝相30℃孵育比酵母相35℃孵育高。结论：申克孢子丝菌的生长形态（菌丝相或酵母相）及孵育温度（30℃或35℃）对其药敏试验MIC值有明显影响。     

动物源性皮肤癣菌病32例患者病原菌体外药敏试验

目的了解动物源性皮肤癣菌病病原菌对常用抗真菌药物的敏感情况。方法收集2013年10月—2014年11月就诊于川北医学院附属医院皮肤科、确诊为动物源性皮肤癣菌病的32例患者的菌株。参照美国临床实验室标准化研究所(CLSI)的M38-A2方案,测定收集的临床菌株对灰黄霉素、伊曲康唑、特比萘芬及氟康唑的最低抑菌浓度(MIC)值。结果灰黄霉素、伊曲康唑、特比萘芬和氟康唑对犬小孢子菌的药敏试验MIC值范围分别为0.125 0～4.000 0μg/ml、 0.031 3～1.000 0μg/ml、0.002 0～0.031 3μg/ml和2.000 0～32.000 0μg/ml,对须癣毛癣菌复合体的药敏试验MIC值范围分别为0.062 5～1.000 0μg/ml、0.0313～0.500 0μg/ml、0.001 0～0.062 5μg/ml和4.000 0～32.000 0μg/ml,对石膏样小孢子菌的药敏试验MIC值范围分别为0.031 3～2.000 0μg/ml、0.062 5～1.000 0μg/ml、0.003 9～0.0625μg/ml和0.500 0～16.000 0μg/ml。结论灰黄霉素、伊曲康唑、特比萘芬和氟康唑对犬小孢子菌、须癣毛癣菌复合体和石膏样小孢子菌的体外药敏试验提示,特比萘芬的抗真菌效果最好,其次为伊曲康唑和灰黄霉素,氟康唑抗真菌效果较差。     

几种抗真菌药物对马尔尼菲青霉的体外药敏试验

目的 观察马尔尼菲青霉（PM）广西野生银星竹鼠寄生株与临床人分离株对伏立康唑和几种常用抗真菌药物的敏感性。方法 采用美国临床实验室标准委员会（CLSI）M27-A2和M38-A方案中的微量稀释法,测定伏立康唑、伊曲康唑、特比萘芬、两性霉素B 及氟康唑对14株广西野生银星竹鼠PM寄生株与25株临床人PM分离株25 ℃菌丝相及37 ℃酵母相的最小抑菌浓度（MIC）。用两样本均数比较t检验比较PM寄生株与临床人PM分离株MIC的差异性,用配对t检验比较同一株菌在两种不同温度相下的MIC差异性。结果 伏立康唑、伊曲康唑、特比萘芬、两性霉素B、氟康唑对PM寄生株菌丝相的MIC分别为：0.0313 ～ 0.1250、0.1250 ～ 1.0000、0.0313 ～ 0.5000、0.2500 ～ 4.0000、2.0000 ～ 8.0000 mg/L;对PM寄生株酵母相的MIC分别为：0.0078 ～ 0.2500、0.0313 ～ 0.5000、0.0313 ～ 1.0000、0.2500 ～ 2.0000、1.0000 ～ 8.0000 mg/L;对PM临床人分离株菌丝相的MIC分别为：0.0313 ～ 0.2500、0.0625 ～ 1.0000、0.0313 ～ 1.0000、0.2500 ～ 4.0000、2.0000 ～ 32.0000 mg/L;对PM临床人分离株酵母相的MIC分别为：0.0039 ～ 0.2500、0.0313 ～ 0.5000、0.0313 ～ 2.0000、0.1250 ～ 2.0000、2.0000 ～ 16.0000 mg/L。5种抗真菌药物对广西野生银星竹鼠PM寄生株与临床人PM分离株菌丝相和酵母相均敏感。同一温度下伏立康唑对两种不同来源PM的MIC最低,其他药物的MIC依次为伊曲康唑、特比萘芬 < 两性霉素B < 氟康唑。同一药物在同一温度下对广西野生银星竹鼠PM寄生株与PM临床分离株的MIC无明显差异;伊曲康唑、两性霉素B、特比萘芬对同一菌株在菌丝相和酵母相下的MIC存在差异。结论 PM对伏立康唑的敏感性最高;来自于广西野生银星竹鼠的PM寄生株与临床人PM分离株对伏立康唑、伊曲康唑、特比萘芬、两性霉素B及氟康唑的MIC类似;菌相的改变可影响PM对伊曲康唑、两性霉素B、特比萘芬的敏感性。     

伊曲康唑治疗孢子丝菌病的疗效及抗真菌药敏试验

目的:观察伊曲康唑对孢子丝菌病的临床疗效,检测孢子丝菌对伊曲康唑的体外敏感性.方法:对48例孢子丝菌病患者给予伊曲康唑胶囊口服治疗;另外30例患者给予碘化钾溶液口服治疗,并采用Etest法检测其中10株孢子丝菌对伊曲康唑的最小抑菌浓度(MIC).结果:伊曲康唑对孢子丝菌病的总有效率91.67％,碘化钾溶液的总有效率86.67％,两组总有效率比较无显著性差异,(P＞0.05).采用Etest法检测10株孢子丝菌对伊曲康唑MIC值,范围为0.012-8μg/mL.结论:虽然伊曲康唑对孢子丝菌病的疗效与碘化钾的疗效相当,但副作用小.Etest法可以作为一种孢子丝菌的体外抗真菌药敏试验方法.     

氟康唑、特比萘芬和伊曲康唑对白念珠菌的体外敏感性试验

目的比较氟康唑、特比萘芬及伊曲康唑对白念珠菌的体外敏感性。方法采用NCCLS公布的M27-A方案微量稀释法测定氟康唑、特比萘芬及伊曲康唑对22株临床分离的白念珠菌的体外敏感性。结果22株白念珠菌对氟康唑耐药9株,敏感12株;对伊曲康唑耐药7株,敏感8株;对特比萘芬耐药12株,敏感3株。结论3种药物中氟康唑的敏感性相对较高,但仍有耐药现象,氟康唑与伊曲康唑存在交叉耐药。     

耐氟康唑的白念珠菌对常见抗真菌药物的敏感性检测

目的测定耐氟康唑白念珠菌对常见抗真菌药物的敏感性,并探讨pH值对微量液基稀释法的影响。方法参照美国临床实验室标准研究所(CLSI)颁布的酵母菌抗真菌药物敏感性试验方案M27-A2,测定氟康唑(FLC)、伏立康唑(VOR)、伊曲康唑(ITC)、两性霉素B(AMB)和米卡芬净(MCFG)的最小抑菌浓度(MIC),并调整RPMI-1640液体培养基的pH值至4.5,测定FLC、VOR和ITC的MIC值。结果FLC的MIC值均>64μg/mL;VOR的MIC值,6株菌>16μg/mL,1株为2μg/mL,3株<1μg/mL;ITC的MIC值,7株≥1μg/mL,3株为0.25～0.5μg/mL;MCFG的MIC值为0.125～0.25μg/mL,AMB的MIC值为1～2μg/mL;使用pH值为4.5的RPMI-1640液体培养基,5种抗真菌药物的MIC值变化不明显。结论10株耐氟康唑的白念珠菌对MCFG均敏感,部分菌株对VOR和(或)ITC交叉耐药;部分耐FLC的白念珠菌对AMB的敏感度有所降低。降低RPMI-1640的pH值,可明显减少拖尾现象,但不影响药敏结果的判定。     

孢子丝菌病药敏和治疗进展

孢子丝菌病是一种深部真菌病,其致病菌申克孢子丝菌为一种双相真菌,双相真菌的体外药敏试验至今仍无统一标准.饱和碘化钾溶液、伊曲康唑、特比萘芬为治疗孢子丝菌病的常用药物,临床中选择合适的抗真菌药物对治疗此病具有重要意义.体外药敏试验是评价抗真菌药物活性的重要方法,也是选择药物的依据之一.     

Sporotrichosis refractory to conventional treatment: therapeutic success with potassium iodide

Sporotrichosis is a subcutaneous mycosis caused by dimorphic fungi of the genus Sporothrix. The authors report a case of fixed cutaneous sporotrichosis with therapeutic failure after 18 months of itraconazole and terbinafine associated with cryosurgery. The patient was cured after the introduction of saturated potassium iodide solution. Sporothrix brasiliensis was the identified species, presenting a susceptibility profile to itraconazole and terbinafine. This fact suggests that therapeutic failure is probably related to the host-fungus interaction rather than drug resistance. It is possible that the immunomodulatory action of the saturated potassium iodide solution may have played an important role in curing this patient.     

In vitro activity of itraconazole in combination with terbinafine against clinical strains of itraconazole-insensitive Sporothrix schenckii

Four strains of Sporothrix schenckii were isolated from patients with cutaneous sporotrichosis who failed itraconazole (ITC) treatment. To investigate the susceptibility of these strains to ITC and terbinafine (TRB) alone and in combination in 2 growth phases in vitro, a checkerboard microdilution method was used in accordance with the recommendations of the Clinical Laboratory Standards Institute (CLSI) was used in our study. The drug interaction was evaluated by assessing the fractional inhibitory concentration index (FICI). The geometric means (GMs) of the minimum inhibitory concentrations (MICs) of 4 insensitive strains were obviously higher than those of the sensitive isolates used for comparison. The FICI analysis revealed that only 2 isolates (25%) were synergistic in yeast form. Our results indicate the failure of clinical treatments might be caused by the insensitivity to ITC of these strains.     

葡萄糖消耗试验用于体外抗真菌药物敏感性试验

目的：评价葡萄糖消耗试验应用于体外抗真菌药物敏感性试验的可行性。方法：分析8种抗真菌药物与40株临床分离菌株在仅含葡萄糖为碳源的酵母氮培养基(YNBG)上孵育后葡萄糖的浓度变化，反映真菌细胞的受抑制程度，同时与NCCLS M27-A液基微量稀释法对比。结果：葡萄糖消耗试验在念珠菌和隐球菌的药敏测定中至少可以节省20小时，对须癣毛癣菌则至少可节省40小时；除两种方法测得的联苯苄唑MIC符合程度较低外，两种方法测得的两性霉素B、特比萘芬、伊曲康唑、酮康唑、咪康唑、氟康唑、环吡酮胺的MIC符合程度均较高。在评价由较低或较高浓度药物诱导的真菌细胞受抑制程度时，葡萄糖消耗试验更精确。结论：葡萄糖消耗试验具有客观量化、快捷、精确的优点，具有推广和应用价值。     

In vitro susceptibility testing of ciclopirox,terbinafine, ketoconazole and itraconazole against dermatophytes and nondermatophytes,and in vitro evaluation of combination antifungal activity

BACKGROUND: With the development of newer antifungal agents with activity against both yeasts and filamentous fungi, there is an increased need to develop and standardize in vitro assays that will evaluate the activity of antimycotics against filamentous fungi. In vitro analysis of antifungal activity of these agents would also allow for the comparison between different antimycotics, which in turn may clarify the reasons for lack of clinical response or serve as an effective therapy for patients with chronic infection. OBJECTIVES: To determine the in vitro susceptibility of fungal organisms to ciclopirox, terbinafine, ketoconazole and itraconazole and to evaluate the in vitro activity and mode of interaction of ciclopirox in combination with either terbinafine or itraconazole. MATERIALS AND METHODS: In the minimum inhibitory concentration (MIC) study 133 strains were evaluated, including dermatophytes (110 strains; 98 from Trichophyton spp.), Candida spp. (14 strains) and nondermatophyte moulds (nine strains). In vitro susceptibility testing was conducted in microbroth dilutions based on the National Committee for Clinical Laboratory Standards (NCCLS) M27-A proposed standard. The testing MIC ranges were 0.003-2 microg mL-1 for ciclopirox and terbinafine, and 0.06-32 microg mL-1 for itraconazole and ketoconazole. For inoculum preparation, dermatophytes were grown on Heinz oatmeal cereal agar slants. Inoculum suspensions of dermatophytes were diluted in RPMI 1640 (Sigma-Aldrich) with the desired final concentration being 2-5 x 103 c.f.u. mL-1. Once inoculated, the microdilution plates were set up according to the NCCLS M27-A method, incubated at 35 degrees C, and read visually following 7 days of incubation. For azole agents, the MIC was the lowest concentration showing 80% growth inhibition; for terbinafine and ciclopirox, the MIC was the lowest concentration showing 100% growth inhibition. In the synergy studies, 29 strains from nondermatophyte species were evaluated using a checkerboard microdilution method. The concentrations tested were: 0 and 0.06-32 microg mL-1 for itraconazole, and 0 and 0.003-4 microg mL-1 for both terbinafine and ciclopirox. Modes of interaction between drugs were classified as synergism, additivism, antagonism or indifference based on fractional inhibitory concentration index values (FIC index). Synergism was defined as an FIC index of < or = 0.50, additivity as an FIC index of < or = 1.0, and antagonism as an FIC index of > or = 2.0. The drug combination was interpreted as indifferent if neither of the drugs had any visible effect on the presence of the other drug. RESULTS: In the MIC study, the dermatophyte MIC values (microg mL-1) (mean +/- SEM) were: ciclopirox (0.04 +/- 0.02), terbinafine (0.04 +/- 0.23), itraconazole (2.28 +/- 7.42) and ketoconazole (0.83 +/- 1.99). The yeast MIC values (microg mL-1) (mean +/- SEM) were: ciclopirox (0.05 +/- 0.02), terbinafine (1.77 +/- 0.58), itraconazole (0.18 +/- 0.27) and ketoconazole (0.56 +/- 0.60). The non-dermatophyte fungi MIC values (microg mL-1) (mean +/- SEM) were: ciclopirox (1.04 +/- 2.62), terbinafine (1.04 +/- 0.95), itraconazole (17.87 +/- 16.75) and ketoconazole (10.69 +/- 13.09). In the synergy study, with ciclopirox in combination with terbinafine, mainly a synergistic or additive reaction was observed; there were no cases of antagonism. For ciclopirox in combination with itraconazole, there were some instances of additivism or synergism, with indifference in the majority of instances; there were no cases of antagonism. CONCLUSIONS: In vitro susceptibility testing indicates that ciclopirox may have a broad antimicrobial profile including dermatophytes, yeasts and other nondermatophytes. Terbinafine is extremely potent against dermatophytes. In vitro evaluation of activity of ciclopirox and terbinafine suggests many instances of synergy or additivism; for ciclopirox and itraconazole there may be indifference, synergy or additivism.     

微量液基稀释法与琼脂稀释法测定马拉色菌体外抗真菌药物敏感性

目的 比较微量液基稀释法与琼脂稀释法检测马拉色菌体外对氟康唑、酮康唑及伊曲康唑敏感性的差异。方法 对临床分离的5种27株马拉色菌,分别用微量液基稀释法与琼脂稀释法测定氟康唑、酮康唑及伊曲康唑对这些马拉色菌的最低抑菌浓度（MIC）。结果 微量液基稀释法显示氟康唑MIC值范围0.25 ～ ≥ 64 mg/L,酮康唑≤0.03 ～ 0.5 mg/L,伊曲康唑≤0.03 ～ 0.125 mg/L;琼脂稀释法显示氟康唑MIC值范围2 ～ ≥ 64 mg/L,酮康唑≤0.03 ～ 0.5 mg/L,伊曲康唑≤0.03 ～ 0.25 mg/L。两种方法测定3种唑类抗真菌药对马拉色菌的活性由高到低为：伊曲康唑、酮康唑、氟康唑。两种方法测得氟康唑、酮康唑及伊曲康唑MIC值符合率分别为78.8%、85.2%、88.9%,组内相关系数分别为0.88、0.80、0.76。结论 氟康唑、酮康唑及伊曲康唑对马拉色菌均有较好的抗菌活性,以伊曲康唑对各菌种的抗菌活性最强。微量液基稀释法与琼脂稀释法相比具有良好的一致性,均适用于马拉色菌体外药敏试验。     

耐伊曲康唑烟曲霉临床分离株对四种抗真菌药物的敏感性测定

目的 测定耐伊曲康唑烟曲霉临床株对4种抗真菌药物的敏感性。方法 自一例对伊曲康唑无效的肺曲霉球患者体内系列分离6株烟曲霉。利用美国临床实验室标准化研究所（CLSI）的微量液基稀释法M38-A方案和E-test法测定6株烟曲霉临床分离株对两性霉素B、卡泊芬净、米卡芬净、伏立康唑和伊曲康唑的敏感性。结果 6株烟曲霉中，2株对伊曲康唑的最低抑菌浓度（MIC）为 0.5 μg/mL，其他4株的MIC均 ＞ 16 μg/mL；两性霉素B和伏立康唑对6株烟曲霉的MIC分别为1 μg/mL和0.25 ~ 1 μg/mL，卡泊芬净和米卡芬净对6株烟曲霉的最低有效浓度均 ≤0.03 μg/mL。E-test法测定结果也显示，卡泊芬净和伏立康唑对6株烟曲霉有良好的抑制活性。结论 耐伊曲康唑烟曲霉临床株对两性霉素B、卡泊芬净、米卡芬净和伏立康唑敏感。