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。     

花斑糠疹与马拉色菌毛囊炎相关危险因素和致病菌分析

目的 研究花斑糠疹和马拉色菌毛囊炎的相关危险因素和致病菌特点,并比较其差异。方法 收集我院临床花斑糠疹和马拉色菌毛囊炎患者,均以1：1设置病例对照组,调查发病相关因素,采用SPSS16.0软件进行单因素和多因素Logstic回归分析。培养阳性菌株采用形态学和生理生化学方法鉴定到菌种,比较菌种分布。结果 共收集到163例花斑糠疹和132马拉色菌毛囊炎患者。单因素回归分析显示与花斑糠疹发病相关的因素有多汗和油性皮肤,与马拉色菌毛囊炎发病相关的因素有男性、潮湿环境、多汗、油性皮肤、刺激性食物、甜腻食物。多因素回归分析显示花斑糠疹的危险因素有多汗和油性皮肤,马拉色菌毛囊炎的危险因素有多汗、油性皮肤、男性和甜腻性食物。两种疾病致病菌均以合轴马拉色菌为优势菌,但糠秕马拉色菌在毛囊炎中占较高比例。结论 多汗和油性皮肤是花斑糠疹和马拉色菌毛囊炎共同的危险因素,而男性、甜腻性食物为马拉色菌毛囊炎独有的危险因素,两种疾病致病分布也存在统计学差异。     

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。     

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。     

随机扩增多态性DNA结合微流芯片鉴定马拉色菌的初步研究

目的 探讨微流芯片在马拉色菌鉴定与分型中的应用优势.方法 收集马拉色菌标准菌株及直接镜检阳性的花斑糠疹患者皮损处皮屑及马拉色菌毛囊炎患者皮损处毛囊内容物培养出的马拉色菌菌株进行DNA测序,鉴定菌种,随机扩增多态性DNA聚合酶链反应(RAPD-PCR)电泳分析及微流芯片基因型定量分析并聚类分析树状图.结果 共分离83株马拉色菌临床菌株,其中72株分离自花斑糠疹,11株分离自马拉色菌毛囊炎.大多数菌株均可被2种随机引物(S22、S24)扩增而获得清晰条带,但以S22引物扩增的条带更为稳定、清晰,作为主要引物.不同种马拉色菌通过微流芯片基因型定量分析得到不同大小的固定阳性条带,所有菌株均可见种间和种内多态性.在DNA测序的基础上,使用RAPD结合微流芯片方法,基本可将8种马拉色菌(糠秕、合轴、球形、厚皮、斯洛菲、日本、大和及皮肤马拉色菌)区别.结论 RAPD结合微流芯片方法作为一种快速、高通量、高灵敏性的分析技术,在马拉色菌种间菌株遗传多样性、亲缘关系的分析及新种鉴定中显示出一定优越性.     

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。     

婴儿皮肤马拉色菌带菌及菌种调查

目的了解1~6个月婴儿皮肤马拉色菌带菌情况和菌种构成及其影响因素。方法采用胶带法粘取1~6个月婴儿额部皮肤处鳞屑,接种于含菜子油培养基进行真菌培养分离马拉色菌,并用生理生化及形态学方法鉴定菌种。结果①75例婴儿皮肤标本有54例培养出马拉色菌,培养阳性率为72%;②共分离出56株马拉色菌,以糠秕马拉色菌、球形马拉色菌、合轴马拉色菌为主;③按月龄、性别、生产方式等因素的不同进行分组,结果不同月龄、性别婴儿皮肤马拉色菌阳性率及菌种构成比较差异均无统计学意义;不同生产方式婴儿马拉色菌培养阳性率比较差异无统计学意义但菌种构成比较差异有统计学意义。结论马拉色菌是婴儿皮肤的常驻菌;婴儿皮肤马拉色菌以糠秕马拉色菌、球形马拉色菌、合轴马拉色菌为主。婴儿皮肤马拉色菌阳性率不受月龄、性别、生产方式等因素的影响,婴儿皮肤马拉色菌菌种构成不受月龄、性别影响。     

马拉色菌属的分类及其在几种皮肤病的分布情况

目的 研究马拉色菌属的分类及其在一些皮肤病的分布情况。方法 以标准株作对照,用生理生化学及形态学方法将１５０株来源于花斑癣（２９株）、马拉色菌毛囊炎（１６株）、脂溢性皮炎（４９株）、甲真菌病（３３株）及正常人皮肤（２３株）的马拉色菌进行分类及描述。并分析了各菌种在一些皮肤病的分布情况。结果 共分离出４个菌种：合轴马拉色菌６５株、康秕马拉色菌１３株、钝形马拉色菌１株、球形马拉色菌７１株。其中合抽马拉     

从包皮龟头炎患者皮损处分离鉴定马拉色菌

目的：了解马拉色菌属各菌种在包皮龟头炎皮损处的构成及其在发病中的作用。方法：从患处取材直接镜检查见马拉色菌孢子和(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。依据生理生化和形念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57例(70．37％)培养并鉴定出马拉色菌(共60株)，其中糠秕马拉色菌20株(33．33％），合轴马拉色菌18株(30．00％)，钝形马拉色菌17株(28．33％)，球形马拉色菌5株(8．33％)。有37例同时分离到念珠菌(其中72．97％为白念珠菌）。44例仅检出马拉色菌的患者中有23例接受抗真菌治疗。结论：糠秕马拉色菌、合轴马拉色菌、钝形马拉色菌是包皮龟头炎患者皮损处的主要菌种；马拉色菌可能单独或与念珠菌协同引起包皮龟头炎。     

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。     

Distribution of Malassezia species in pityriasis versicolor and seborrhoeic dermatitis in Greece. Typing of the major pityriasis versicolor isolate M. globosa

BACKGROUND: The expansion of the genus Malassezia has generated interest in the epidemiological investigation of the distribution of new species in a range of dermatoses, on which variable results have been reported from different geographical regions. No data are thus far available from South-east Europe (Greece). OBJECTIVES: To study the distribution of Malassezia species in pityriasis versicolor (PV) and seborrhoeic dermatitis (SD) and to investigate whether polymorphisms in the internal transcribed spacer (ITS) 1 region facilitate detection of M. globosa and M. sympodialis subtypes. METHODS: In total, 109 patients with PV and SD and positive Malassezia cultures were included in the study. Age, gender, primary/recurrent episode, disease extent and clinical form of PV were recorded. ITS 1 polymorphisms of M. globosa and M. sympodialis type and clinical strains were investigated by polymerase chain reaction (PCR)-single-strand conformational polymorphism (SSCP) analysis. RESULTS: Malassezia globosa was the prevalent species isolated from PV and SD either alone (77% and 39%, respectively) or in combination (13% and 18%, respectively) with other Malassezia species. The pigmented form of PV was strongly correlated with the female gender. PCR-SSCP differentiated five subgroups of M. globosa with one being associated with extensive clinical disease. All M. sympodialis isolates displayed a homogeneous ITS 1 PCR-SSCP profile. CONCLUSIONS: Malassezia species isolation rates were in agreement with those reported from South-west Europe. PCR-SSCP of the ITS 1 is useful for highlighting prospective clinical implications of M. globosa subtypes.     

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。     

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.     

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   

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.     

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。