真菌病

20 0 4 0 0 91　从包皮龟头炎患者皮损处分离鉴定马拉色菌 /郭晓莉 (四川大学华西医院皮肤科 )…∥临床皮肤科杂志 .- 2 0 0 3,32 (7) .- 385～ 387从患处取材直接镜检见马拉色菌孢子和 (或 )菌丝的包皮龟头炎患者作为研究对象。用胶带法取材后分别接种在含菜子油的培养基及无放线菌酮的沙堡培养基分离菌种。依据生理生化和形态学特点及转种到科玛嘉显色培养基和米粉吐温80琼脂培养结果鉴定出马拉色菌和 (或 )念珠菌。结果 :81例患者中有 5 7例培养并鉴定出马拉色菌 (共 6 0株 ) ,其中糠秕马拉色菌 2 0株 ,合轴马拉色菌 18株 ,钝形马拉色菌 …     

球形马拉色菌是马拉色菌毛囊炎患者皮损毛囊内的主要菌种

目的 确定引起马拉色菌毛囊炎的主要菌种,比较皮损毛囊内和毛周表面皮肤菌种是否一致,患者性别、年龄、病情与分离菌种的关系。方法 菜子油培养基培养,根据形态及生理生化特点进行鉴定。结果 从毛周表面皮肤分离的319株菌中鉴定出合轴马拉色菌247株(77.43%)、糠秕马拉色菌40株(12.54%)、球形马拉色菌27株(8.46%)、钝形马拉色菌5株(1.57%);从毛囊内分离的314株菌中鉴定出球形马拉色菌252株(80.25%)、合轴马拉色菌57株(18.15%)、糠秕马拉色菌4株(1.27%)、钝形马拉色菌1株(0.32%),菌种构成差异有显著性(P<0.01)。毛囊内菌种构成与患者的性别和年龄有关,与病情无关。毛囊内和毛周表面皮肤同时阳性的279株中菌种不一致204株,菌种一致75株。结论 马拉色菌毛囊炎毛囊内的主要菌种为球形马拉色菌,其表面皮肤则以合轴马拉色菌为主。     

南通和南京马拉色菌毛囊炎临床调查及致病菌种分析

目的 探讨南通和南京马拉色菌毛囊炎的易感因素及致病菌种在不同地区、不同部位马拉色菌毛囊炎中的菌种分布情况。方法 对符合病例收集纳入标准的患者进行问卷调查,取毛囊内容物进行真菌涂片、培养;并根据形态学和生理生化特征进行菌种鉴定。结果 241例临床诊断为马拉色菌毛囊炎的患者,真菌涂片204例阳性,涂片阳性率84.65%;收集标本259份,共得阳性株213株,其中马拉色菌209株,念珠菌4株（占1.54%）,真菌培养阳性率82.24%。菌种鉴定：209株马拉色菌活化菌种后,可供鉴定的马拉色菌菌株186株,共检测到6个菌种的马拉色菌,其中糠秕马拉色菌111株（59.68%）、斯洛菲马拉色菌43株（23.12%）、合轴马拉色菌17株（9.14%）、球形马拉色菌9株（4.84%）、厚皮马拉色菌4株（2.15%）、钝形马拉色菌2株（1.08%）。不同个体、不同部位的菌种分布：胸部、后背、腹部和面颈部以糠秕马拉色菌为主,上肢、肩部和头顶以斯洛菲马拉色菌为主,下肢均为球形马拉色菌。同一个体、不同发病部位存在不同的菌种,主要为糠秕马拉色菌合并合轴或斯洛菲马拉色菌。 结论 南通和南京马拉色菌毛囊炎存在6种马拉色菌致病菌种,糠秕和斯洛菲马拉色菌是主要的致病菌种。     

从花斑癣患者的皮损区及非皮损区分离和鉴定马拉色菌

目的 研究花斑癣患者皮损区及非皮损区马拉色菌菌种构成;不同解剖部位、皮损颜色及各菌种的分布情况;患者病情和年龄与菌种构成的关系。方法 用无菌胶带粘取113例花斑癣患者皮损区及非皮损区共629个部位的皮屑,分别接种于含菜子油培养基中分离马拉色菌,用生理生化及形态学方法鉴定菌种。结果 皮损区与相对应的非皮损区马拉色菌分离阳性率无差别,非皮损区额部和胸背部分离阳性率高于上、下肢。共分离到565株马拉色菌,鉴定出合轴马拉色菌(44.78%)、糠秕马拉色菌(32.94%)、球形马拉色菌(11.68%)、钝形马拉色菌(5.84%)及限制马拉色菌(4.76%)共5个种,有27处(5.01%)同时分离到两种菌。皮损区与非皮损区菌种构成无明显差别,限制马拉色菌主要从额部分离出。菌种构成与皮损面积无关,但与皮损颜色和患者年龄有关。皮损颜色与病程无关。结论 花斑癣患者皮损区与非皮损区马拉色菌分离阳性率和菌种构成基本一致,与病情无关,而不同解剖部位、皮损类型及年龄患者的菌种构成有一定差异。     

花斑糠疹和马拉色菌毛囊炎菌种分布特点分析

目的探讨花斑糠疹和马拉色菌毛囊炎的菌种分布特点。方法从临床诊断为花斑糠疹和马拉色菌毛囊炎的患者分离培养菌种,通过形态学和生理生化学方法鉴定菌种,并比较两组患者菌种分布情况。结果共收集花斑糠疹病例161例,培养阳性121株,其中合轴马拉色菌46株,糠秕马拉色菌13株,球形马拉色菌29株,钝性马拉色菌33株;马拉色菌毛囊炎135例,培养阳性114株,其中合轴马拉色菌52株,糠秕马拉色菌43株,球形马拉色菌13株,钝性马拉色菌6株。两种疾病菌种分布差异有显著性(P<0.005)。结论花斑糠疹和马拉色菌毛囊炎菌种分布存在差异,可能为两种疾病不同临床表现的原因之一,但结果尚需慎重解释,需要分子水平的进一步研究。     

无锡和南京地区汗斑病原菌种的分布

探讨汗斑患者马拉色菌的菌学特征和分布取临床标本做镜检和培养，观察菌的形态学特征，并采用一系列牛化鉴定方法鉴定菌种。各菌种菌落形态不同，特别是钝形马拉色菌和球形马拉色菌菌落形态特征明显，孢子形态具有特征性，容易鉴别，可作为鉴定指标之一；致病优势菌依次为糠秕马拉色菌、合轴马拉色菌，球形马拉色菌等。准确鉴定屿拉色菌需结合形态学和生化特征；7个致病菌种在汗斑皮损均有分布。     

花斑糠疹和马拉色菌毛囊炎致病菌菌种分析

目的使用限制性内切酶方法鉴定临床花斑糠疹和马拉色菌毛囊炎致病菌菌种分布并比较其差异。方法收集临床花斑糠疹和马拉色菌毛囊炎标本,植于Leeming-Notman培养基。培养阳性菌株提取DNA,扩增其26srDNA片段,用CfoⅠ酶和BstF51酶切。结果共鉴定花斑糠疹阳性菌株128份,其中糠秕马拉色菌49份,合轴马拉色菌23份,球形马拉色菌26份,钝性马拉色菌6份,M.japponica 1份,斯洛菲马拉色菌1份,混合感染标本22份。共鉴定马拉色菌毛囊炎阳性标本70份,其中糠秕马拉色菌43株,合轴马拉色菌5株,球形马拉色菌9株,钝性马拉色菌1株,混合感染标本12份。两种疾病菌种分布差异存在统计学意义(P=0.009)。结论花斑糠疹和马拉色菌毛囊炎致病菌菌种存在一定差异。限制性酶切方法为一种准确、快速的马拉色菌菌种鉴定方法。     

尿素液基微量稀释法用于马拉色菌的敏感性试验

马拉色菌属最近被重新分为 7个种 ,糠秕马拉色菌、厚皮马拉色菌、合轴马拉色菌、球形马拉色菌、钝形马拉色菌、限制性马拉色菌、斯洛菲马拉色菌。1992年 ,ＮＣＣＬＳ推荐对酵母菌敏感试验的液体微量稀释法。然而 ,这种方法对马拉色菌属 (厚皮马拉色菌除外 )并不适用 ,因为马拉色菌在无脂质的培养基上不生长。马拉色菌被证实和新生隐球菌一样 ,尿素酶阳性。作者试图建立一个实用的敏感性试验方法去获得多种药物对具有尿素酶活性的 7种马拉色菌抗菌活性的有用资料。试验菌株共 4 2株 (13株糠秕马拉色菌、1株球形马拉色菌、6株钝形马拉色菌、1…     

从花斑糠疹皮损分离的马拉色菌随机扩增多态性DNA研究

目的 用随机扩增多态性DNA方法研究分离自花斑糠疹的马拉色菌种间和株间差异,了解随机扩增多态性DNA分析（RAPD）与生理生化方法在菌种分型上的差异及菌株DNA型别和菌种间的关系。方法 用氯化苄法提取马拉色菌标准株（10株7个种）和临床分离株（47株）的基因组DNA,其中临床株分离自34例花斑糠疹患者,经形态学和生理生化方法鉴定为5个种（合轴马拉色菌、糠秕马拉色菌、钝形马拉色菌、球形马拉色菌、限制马拉色菌）,用4种随机引物（S22、S24、S25、S33）对菌株DNA做PCR随机扩增,NTSYS软件自动生成树状分支图。结果 绝大多数标本均可被4种引物扩增而获得清晰条带,其中2种引物（S22、S24）的条带更为稳定、清晰。共82条DNA片段被扩增,所有菌株均可见种间和株间多态性。有4例患者皮损同时分离出不同种的菌株显示遗传相似性高,在树状图中归入一类。结论 来自同一宿主的不同菌株遗传趋同现象提示马拉色菌的种特异性、菌种演化与宿主间存在密切关系。     

从夫妻双方的头皮屑中分离和鉴定马拉色菌

目的 探讨夫妻双方的头皮屑中马拉色菌的带菌情况，菌种构成及夫妻间共用梳子与马拉色菌菌种一致性的关系，方法 采用Leeming和Notman培养基，Dixon培养基和含菜子油培养基分别同时培养114对夫妻的头皮屑，观察培养阳性率与头皮屑严重程度的关系。根据生理生化学及形态学特点鉴定菌种，并比较在这3种培养基中菌落初长时间及对菌种有无选择性。结果 从114对夫妻（共228人0中153人的头皮屑中分离到马拉色菌，阳性率为67.11%。培养阳性率与头皮屑的严重程度有正相关关系，3种培养基共分离到马拉色菌，阳性率为67.11%。培养阳性率与头皮屑的严重程度有正相关关系。3种培养基共分离到459株马拉色菌，从中鉴定出合轴马拉色菌（33.33%)，球形马拉色菌（25.05%)，限制马拉色菌（15.47%)，糠秕马拉色菌（13.73%)和钝形马拉色菌（12.42%)共5个种，3种培养基对菌种无选择性，但在含菜子油的培养基中菌落初长时间最短，同一头皮屑在3种培养基上分离到同一菌种有129人（56.58%)，分离到2种菌的有24人（10.53%)，共用梳子的夫妻双方的头皮屑中菌种相同者（35.96%)显著多于菌种不同者（13.16%)；夫妻间共用梳子者头皮屑中菌种相同者（50.67%)显著多于非共用梳子者（7.69%)。结论 头皮屑中的优势菌种主要为合轴和球形马拉色菌，夫妻双方的头皮屑中马拉色菌菌种一致率较高，而夫妻间共用梳子组的菌种一致率更高，提示马拉色菌可在夫妻之间传播，共用梳子可能是重要的传播媒介。     

Effects of topical vehicles on growth of the lipophilic Malassezia species

In the present study, the abilities of major Malassezia species, M. sympodialis, M. globosa and M. furfur, to assimilate topical agents, which have been widely used as a material of ointment for skin diseases, were tested. Obvious growth of M. furfur on GYEP agar plate was noted in the presence of white petrolatum, purified white petrolatum, hydrophilic ointment and heparinoid in hydrophilic ointment, and also M. sympodialis showed similar growth when they were cultured with hydrophilic or heparinoid in hydrophilic ointment. In contrast, M. globosa did not grow on GYEP in the presence of the any topical agents tested. These results suggest that Malassezia species, especially M. furfur and M. sympodialis, assimilate several topical agents and showed the drug-depended cell growth.     

Differentiation of Malassezia species: selectivity of Cremophor EL, castor oil and ricinoleic acid for M. furfur

In recent years, the genus Malassezia has been reclassified based on molecular data. In addition to M. furfur , M. pachydermatis and M. sympodialis , four new species, M. globosa , M. obtusa , M. restricta and M. slooffiiae , have been described. However, apart from their lipid dependence, little is known about the metabolism and nutritional requirements of all the seven species. Further to recent studies, 10 hydrophilic emulsifiers (HLB > 10) were examined in an agar diffusion test to determine their growth-promoting effect on reference strains of the different Malassezia species. Polyethylene glycol (PEG) 7 glyceryl monoalcanoate (Cetiol HE), PEG–glyceryl stearate (Tagat S2) and macrogol-50 stearate (Myrj 53) were metabolized by all strains, while PEG-35 castor oil (Cremophor EL) was metabolized only by M. furfur . The latter observation is due to a different metabolism of castor oil and its main component, ricinoleic acid (12-hydroxy oleic acid), which may also give an insight into the pathogenesis of diseases that are associated with Malassezia spp. As hydroxy fatty acids are important in maintaining the epidermal structure and function, their metabolism specifically by M. furfur might clarify some clinical aspects of pityriasis versicolor. Apart from this speculation, use of Cremophor EL, with splitting of esculin as an additional key character, improves the distinction of the species M. furfur , M. slooffiae and M. sympodialis .     

In vitro susceptibility of the seven Malassezia species to ketoconazole, voriconazole, itraconazole and terbinafine

Fifty-five strains, either authentic or ex-type, of seven Malassezia species were investigated for in vitro susceptibility to various concentrations (0.03-64.0 microg/mL) of three azole drugs, ketoconazole, voriconazole and itraconazole, as well as the allylamine terbinafine, using the agar dilution method. All strains of the seven Malassezia species were susceptible to the three azole drugs at low concentrations. M. furfur, M. sympodialis, M. slooffiae, M. pachydermatis, M. globosa, M. obtusa and M. restricta were most sensitive to ketoconazole and itraconazole, with minimum inhibitory concentrations (MICs) ranging from < or = 0.03 to 0.125 microg/mL. The recently introduced antifungal, voriconazole, was also very effective, with MIC80 values < or = 0.03 microg/mL for 80% of strains. MICs of terbinafine against the seven Malassezia species ranged from 相似文献   

IgE antibodies to Malassezia furfur, M. sympodialis and Pityrosporum orbiculare in patients with atopic dermatitis, seborrheic eczema or pityriasis versicolor, and identification of respective allergens

Malassezia yeasts may be a trigger factor for atopic dermatitis. Following the recent reclassification of the genus, the presence of specific IgE antibodies was examined in the sera of patients with atopic dermatitis (n = 223), pityriasis versicolor (n = 83), seborrheic eczema (n = 50) and hymenoptera allergy (n = 39) and in controls without skin diseases (n = 50). In addition to using the commercially available radioallergosorbent test (RAST) for Pityrosporum orbiculare couplings were also made against the reference strains for M. furfur and M. sympodialis. To characterize the specificity and molecular weight of corresponding epitopes identical material was used for production of an immunoblot. Despite high total levels of IgE, controls and patients with pityriasis versicolor showed no specific IgE antibodies. Six patients (12%) with seborrheic eczema were positive while 78 patients (35%) with atopic dermatitis had specific IgE antibodies in higher RAST classes that differed between the Malassezia species. The molecular weights of the main antigens of M. sympodialis and M. furfur were determined to be 15, 22, 30, 37, 40, 58, 79, 92, 99 and 124 kDa and 15, 25, 27, 43, 58, 92, 99 and 107 kDa, respectively. Evaluated according to the location of their disease, patients with head and neck lesions most frequently showed Malassezia-specific IgE antibodies. However, there were differences between the Malassezia species tested, the previously used strain P. orbiculare being assignable to the species M. sympodialis.     

AhR ligands, malassezin, and indolo[3,2-b]carbazole are selectively produced by Malassezia furfur strains isolated from seborrheic dermatitis

Malassezia yeasts are connected with seborrheic dermatitis (SD) whereas M. furfur pathogenicity is associated with the production of bioactive indoles. In this study, the production of indoles by M. furfur isolates from healthy and diseased skin was compared, the respective HPLC patterns were analyzed, and substances that are preferentially synthesized by strains isolated from SD lesions were isolated and characterized. Malassezin, pityriacitrin, indole-3-carbaldehyde, and indolo[3,2-b]carbazole (ICZ) were isolated by HPLC from extracts of M. furfur grown in L-tryptophan agar, and identified by nuclear magnetic resonance and mass spectroscopy. Of these, ICZ, a potent ligand of the aryl hydrocarbon receptor (AhR), is described for the first time to our knowledge as a M. furfur metabolite. HPLC-photodiode array detection analysis of strain extracts from 7 healthy subjects and 10 SD patients showed that M. furfur isolates from only SD patients consistently produce malassezin and ICZ. This discriminatory production of AhR agonists provides initial evidence for a previously unreported mechanism triggering development of SD and indicates that the variable pathogenicity patterns recorded for M. furfur-associated SD conditions may be attributed to selective production (P<0.001) of measurable bioactive indoles.     

Antigenic components of Malassezia species for immunoglobulin E antibodies in sera of patients with atopic dermatitis

Antigenic components of Malassezia furfur, M. globosa, M. restricta, M. slooffiae, and M. sympodialis were studied for immunoglobulin E antibodies in sera of patients with atopic dermatitis (AD). Antigenic components were extracted from Malassezia cells by treatment with beta-mercaptoethanol, referred to as 2-ME extract. CBB staining and lectin blots using Con A, LCA, PHA-E4, PNA or RCA120 showed that the 2-ME extracts contained several species-dependent components that differed quantitatively and qualitatively among the Malassezia species at the protein level. In the Western blot with the 2-ME extracts, of 54 sera of the patients with AD (54 patients), the patients' IgE antibodies most frequently recognized components showing molecular weights of 43-46 kDa for M. slooffiae, 12-22 kDa for M. sympodialis, 35-40 kDa for M. restricta, 45-50 kDa for M. globosa, and of 67-72 kDa for M. furfur, respectively. In the correlative study, in which the total band intensities generated for each extract in Western blot were compared among the Malassezia species, the intensity for M. globosa was well correlated with that for M. sympodialis (r=0.756). In the Western blot inhibition test, the 2-ME extract of M. globosa partially inhibited the reaction of the antigenic components of other Malassezia species with the patient's IgE antibodies. These results indicated that Malassezia species contained both species-specific and common antigenic components at the IgE antibody level.