红色毛癣菌菌株的特异性PCR鉴定方法研究

目的建立一种快速的红色毛癣菌分子生物学鉴定方法。方法根据红色毛癣菌保守区域-真菌核糖体DNA（rDNA）的转录间隔区（ITS）设计特异性引物,采用上游：ITS19865＇GAC ACC AAG AAA AAA TTC TCT GAA GA3＇,下游：ITS24415＇GTC CTG AGG GCG CTG AA3＇为引物对45株红色毛癣菌、5株须癣毛癣菌和1株紫色毛癣菌菌株的DNA进行PCR扩增,观察产物电泳带型的差异。结果 45株红色毛癣菌均能扩增出目的片段,5株须癣毛癣菌和1株紫色毛癣菌均无目的片段扩增出。结论红色毛癣菌可用特异引物PCR方法快速鉴定。     

家庭内红色毛癣菌病患者间菌株差异性分析

目的用PCR技术比较分离自同一家庭红色毛癣菌病患者的菌株差异性,分析家庭内多发的红色毛癣菌病的致病菌株是家内相互感染,还是家外感染。方法以家庭内多发的皮肤癣菌病患者为研究对象,分离致病菌株并以传统方法鉴定菌种。再分别用随机扩增多态性DNA(RAPD)和巢式PCR特异扩增红色毛癣菌的串联重复亚元件(TRSS:TRS-1/TRS-2)产生的指纹图谱分析种内株间有无差异性。结果纳入实验的16株菌分离自8个家庭,用形态学等方法及种特异引物均鉴定为红色毛癣菌。RAPD显示4个家庭内的菌株间有差异性,TRS-1区PCR指纹图谱显示5个家庭内菌株有株间差异,TRS-2区能鉴定出2个家庭内菌株间有差异。综合各方法共区分出6个家庭内的菌株间有带型差异。结论该研究提示家庭内多发红色毛癣菌病从家外途径感染率高于家内感染。TRS-1区PCR指纹图谱对红色毛癣菌的菌株区分度高于RAPD,更适于红色毛癣菌株间分型。结合多种分子分型方法可最大限度发现不同菌株间的差异。     

我国代表地区须癣毛癣菌复合体的分子鉴定与分型研究

目的对我国代表地区的须癣毛癣菌菌株进行分子再鉴定和分型研究。方法选取我国南北方8个省市地区经表型鉴定的须癣毛癣菌菌株47株,通过再培养形态观察、生理试验;PCR扩增核糖体DNA(rDNA)的内转录间隔区(ITS)和核糖体大亚基(LSU)D1-D2区,测序后利用数据库进行序列比对,对须癣毛癣菌复合体进行再鉴定;PCR扩增rDNA非转录间隔区(NTS)的三个串联重复亚单位S0、S1和S2区,进行种内分型,并比较不同部位来源菌株型别的差异性。结果我国南北方8个省市地区47株须癣毛癣菌中3株鉴定为断发毛癣菌,6株鉴定为无性型苯海姆节皮菌,其余均鉴定为万博节皮菌中的亲人型趾间毛癣菌;三对不同引物扩增38株趾间型毛癣菌和2株苯海姆节皮菌NTS区,共产生28种特征性带型。带型和菌株来源及发生部位无相关性。结论我国分离自人类须癣毛癣菌复合体的主要组成菌种为趾间毛癣菌;ITS区结合LSU D1-D2区测序有助于鉴定须癣毛癣菌复合体至种水平;NTS区的三个串联重复亚单位所产生的特征性指纹图提供了一种快速、稳定的分子生物学种内分型方法,可应用于趾间毛癣菌感染的流行病学研究。     

须癣毛癣菌的形态学及分子生物学鉴定

目的对临床上分离自人(36株)及狐狸(5株)的须癣毛癣菌菌株进行新的分类系统鉴定,并检测传统的分类方法是否能满足临床鉴定需要。方法①观察原鉴定为须癣毛癣菌菌株在沙氏培养基、1%蛋白胨培养基、溴甲酚紫乳固体葡萄糖琼脂培养基(BCP-MSG)和显微镜下形态学及尿素酶、毛发穿孔等生理学试验表现。②通过ITS区段和LSU区段分子生物学序列分析进行新的分类系统的菌种分型,并对传统形态学和生理学鉴定方法进行检测。结果①41株须癣毛癣菌形态学及生理学试验符合须癣毛癣菌(38株)和红色毛癣菌(3株)菌落的特点。②ITS区段序列分析发现ITS区段能将须癣毛癣菌和红色毛癣菌准确的鉴定到种,但无法明确其种内分型;而LSU区段序列分析可对36株(36/38)须癣毛癣菌有性型做出明确的鉴定。结论传统实验室鉴定方法仍具有其有效性及可靠性。通过分子生物学鉴定,临床分离的须癣毛癣菌皆为指(趾)间毛癣菌(38/38),而LSU区段的序列分析鉴定狐狸源性菌株皆属于本海姆节皮菌,有别于大多数人源性菌株有性型为万博节皮菌,对于须癣毛癣菌的菌种鉴定更优于ITS区段,但分子生物学试验还需结合形态学的观察,才能够对菌种做出正确的鉴定。     

106株红色毛癣菌基因分型与药敏分析CSCD

目的探索海南地区的红色毛癣菌基因型与感染部位、药敏的关系。方法基于红色毛癣菌核糖体rDNA非转录区(NTS)的基因分型进行种内分型,依据“CLSI-M38-A2”方案进行药敏实验。结果106株红色毛癣菌TRS-1基因型有5种带型,其中TypeⅠ52株(49.06%),TypeⅡ14株(13.21%),TypeⅢ5株(4.72%),TypeⅣ型3株(2.83%),其他带型32株(30.18%)。TRS-2基因型有3种带型,其中TypeⅠ68株(64.15%),TypeⅡ6株(5.66%),其他带型32株(30.19%)。药敏结果MIC几何均数由低至高分别为特比萘芬(0.0092μg/mL)、伏立康唑(0.0181μg/mL)、伊曲康唑(0.1491μg/mL)、酮康唑(0.1630μg/mL)、氟康唑(2.3164μg/mL)。有42株菌表现出对抗真菌药物不敏感,不敏感菌株TRS-1和TRS-2分型均以TypeⅠ为主。结论海南地区流行的红色毛癣菌TRS-1和TRS-2基因型均以TypeⅠ为主。以对特比萘芬(0.0092μg/mL)、伏立康唑(0.0181μg/mL)MIC几何均数最低。本结果发现本地区的红色毛癣菌不敏感菌株与基因带型关系不大,也与感染部位无关,可能与来源有关。     

297例浅部真菌病临床及病原菌菌种分析

目的 探讨皮肤浅部真菌病致病真菌菌种的构成.方法 对297例真菌涂片阳性和培养阳性的浅部真菌病患者,取标本进行分离培养及菌种鉴定,培养阳性标本在形态学上不能准确鉴定的,进行梅里埃API 20C AUX酵母菌鉴定试剂盒或核糖体DNA （rDNA） ITS区序列测定,确切鉴定菌种.使用SPSS 17.0统计软件对于结果进行统计分析.结果 共分离培养出致病菌13种,其中红色毛癣菌86株（29.0％）,须癣毛癣菌68株（22.9％）,念珠菌属59株（19.9％）,暗色真菌属13株（4.4％）,曲霉菌属13株（4.4％）,红酵母菌12株（4.0％）,青霉菌属9株（3.0％）,毛霉菌9株（3.0％）,犬小孢子菌5株（1.7％）,浅白隐球菌3株（1.0％）,毛孢子菌属2株（0.7％）,絮状表皮癣菌1株（0.3％）,混合感染17株（5.7％）.结论 本地区浅部真菌病以甲癣为主,主要致病真菌是红色毛癣菌,但其他种类真菌感染尤其是念珠菌属有明显上升趋势.     

红色毛癣菌致脓癣1例

目的报告1例由红色毛癣菌引起的脓癣,通过病发分子鉴定明确病原菌,早期诊断并进行经验性治疗。方法提取病发及脓液中的真菌DNA,进行ITS区扩增、测序和比对;同时,对病发和脓液进行真菌镜检及培养,对培养阳性菌落进行形态学、分子生物学鉴定以及体外药敏检测。结果病发和脓液DNA提取及ITS区鉴定提示红色毛癣菌;真菌镜检阴性;真菌培养经形态学和分子生物学鉴定提示红色毛癣菌。体外药物敏感性检测显示其对特比萘芬高度敏感。诊断为红色毛癣菌所致头癣,予口服特比萘芬125 mg/d,治疗1个月后明显改善。结论直接提取病变组织DNA进行早期分子诊断能够能尽早明确致病菌种并指导用药。     

皮肤癣菌体外蛋白水解酶活性测定

目的观察皮肤癣菌的体外蛋白水解酶活性;比较分离自不同感染部位的红色毛癣菌的体外蛋白水解酶活性。方法实验菌株包括来自不同感染部位的红色毛癣菌22株、须癣毛癣菌3株、犬小孢子菌5株,进行体外培养,并利用9-羟基乙酚噻唑标识的酪蛋白和酶标仪检测真菌细胞外蛋白水解酶的活性。结果须癣毛癣菌的体外蛋白水解酶活性高于红色毛癣菌和犬小孢子菌（P〈0.05）,而红色毛癣菌和犬小孢子菌之间无差异（P〉0.05）。红色毛癣菌的细胞外蛋白水解酶活性在分离自浅部感染部位的菌株之间无差异（P〉0.05）,但高于引起毛癣菌肉芽肿的菌株（P〈0.05）。结论不同的皮肤癣菌体外蛋白水解酶活性可能不同;分离自不同感染部位的同一菌种的体外蛋白水解酶活性也有可能不同。     

85株皮肤癣菌感染的病原菌分析及标准化体外药物敏感性研究

目的分析深圳地区教学医院门诊皮肤癣菌感染的病原菌特点,测定7种抗真菌药物对85株皮肤癣菌的体外抗真菌活性。方法收集疑诊为皮肤癣菌感染的患者的皮屑(或甲板刮屑、毛发)进行KOH直接镜检和培养。并通过测定皮肤癣菌临床分离菌株的核糖体DNA内转录间隔区(ITS)区序列鉴定菌种。参考CLSI M38-A2方案,测定7种抗真菌药物对85株鉴定为皮肤癣菌的临床分离菌株的体外抗真菌活性。结果 161例疑诊为皮肤癣菌感染的患者中KOH直接镜检皮肤癣菌的阳性率为67.7%,真菌培养皮肤癣菌的阳性率为52.8%。菌种鉴定结果为红色毛癣菌68例、犬小孢子菌7例、石膏样小孢子菌3例、趾间毛癣菌5例、紫色毛癣菌1例、断发毛癣菌1例。体外药敏试验显示特比萘芬(GM MIC,0.032μg·mL~(-1), MIC范围, 0.001～0.125μg·mL~(-1))对6种皮肤癣菌表现出良好的体外抗真菌活性,而伏立康唑(GM MIC, 0.041μg·mL~(-1), MIC范围, 0.032～0.125μg·mL~(-1))在唑类药物中的体外MIC值相对较低。结论本研究中皮肤癣菌感染的致病菌以红色毛癣菌为主。标准化药敏试验证实特比萘芬和伏立康唑对分离的皮肤癣菌具有较强体外抗菌活力,适用于本地区皮肤癣菌感染的治疗。     

安徽省土壤绿僵菌分离株的分子鉴定

目的进一步验证ITS序列的系统发育分析可为绿僵菌属种的鉴定提供重要的参考依据。方法对分离自安徽土壤的13株绿僵菌菌株的内转录间隔区（ITS）片段进行PCR扩增和序列测定,采用Blast方法将测序结果在GenBank中进行同源搜索,依据邻接法构建获得与其相关菌株的ITS序列系统发育树。结果供试菌株分别位于系统发育树的3个分支上,分支I包括8个菌株和金龟子绿僵菌小孢变种,1个菌株和金龟子绿僵菌鳞鳃金龟变种形成分支III,另外4个菌株和黄绿绿僵菌棉蚜变种聚为分支X。结论结合同源比较的数据,将这8个、4个和1个绿僵菌菌株分别鉴定为金龟子绿僵菌小孢变种、黄绿绿僵菌棉蚜变种和金龟子绿僵菌鳞鳃金龟变种。     

Epidemiology of dermatophytoses: retrospective analysis from 2005 to 2010 and comparison with previous data from 1975

Dermatophyte infections are extremely frequent worldwide and their epidemiological features vary according to the geographical area and have changed in the last decades. We studied the spectrum of dermatophytoses by means of a retrospective analysis involving 6,133 patients referred to the Mycology Service of the Dermatology Clinic of Policlinico Hospital - University of Bari, Italy during the period 2005-2010. The most frequent clinical forms were tinea unguium (39.2% of the total dermatophytoses), tinea corporis (22.7%) and tinea pedis (20.4%). There was a predominance of women for tinea unguium and corporis and of men for tinea pedis and especially tinea cruris. T. rubrum was the prevalent causative agent, implicated in 64% of total cases, followed by M. canis (14%) and T. mentagrophytes (10%). The retrospective evaluation of epidemiological data collected at our Clinic since 1975 showed a gradual decrease in the frequency of tinea cruris, tinea corporis, and tinea capitis over time. On the contrary, during the past two decades, there has been a progressive increase in the frequency of tinea pedis and especially of tinea unguium. In parallel with this changing pattern, the frequency of isolation of T. rubrum has shown a continuous increase during the last 35 years, whereas a progressive decline of the etiological role of T. violaceum, M. canis and even more of E. floccosum has been noted.     

无锡地区293例儿童浅部真菌病临床分析

目的研究无锡地区儿童浅部真菌病的流行及病原菌特点。方法采用真菌镜检和含氯霉素(和含氯霉素及放线菌酮)的葡萄糖蛋白胨琼脂培养,根据菌落形态及其镜下形态鉴定菌种。结果 293例临床诊断患儿中,直接镜检阳性245例,阳性率83.6%,其中皮肤念珠菌病、足癣、头癣发病例数居前3位,分别为79例(32.2%)、50例(20.4%)和46例(18.8%),共175例,占总病例数的71.4%,其次分别为体癣、甲真菌病、手癣、股癣、花斑糠疹,分别为29例(11.8%)、13例(5.3%)、11例(4.5%)、10例(4.1%)、7例(2.9%)。随着年龄增长,患儿易患疾病有所差异,花斑糠疹0～2岁发病率最高;皮肤念珠菌病0～3岁发病率最高;头癣4～8岁发病率最高;体股癣、手、足癣9～15岁发病率最高。238例镜检阳性标本(7例花斑糠疹除外)共培养出致病菌145株,阳性率为60.9%,其中皮肤念珠菌病培养阳性率为70.9%,股癣70%,体癣51.7%,甲真菌病53.8%,头癣65.2%,足癣50%,手癣45.5%。在145株致病菌中,念珠菌属63例(43.45%),红色毛癣菌43例(29.7%),犬小孢子菌26例(17.93%),须癣毛癣菌4例(2.76%),紫色毛癣菌3例(2.07%),石膏样小孢子菌3例(2.07%),断发毛癣菌2例(1.38%),絮状表皮癣菌1例(0.69%)。结论本研究为无锡地区防治儿童浅部真菌病提供了依据。     

New data on the epidemiology of Tinea cruris in Israel

Among 250 patients with tinea cruris (160 males, 90 females) lesions of both tinea pedis and tinea unguium were found in 221 and lesions of only tinea pedis in another 25. The four children were free of associated dermatophytic lesions. The predominant causative agent was T. rubrum, which was also found to be responsible for the hairy area invasion of the beard and pubis in 12 patients and for the widespread lesions of tinea corporis in 10 patients with lymphoma. None of these patients had noticed the discrete nail and foot involvement. It is therefore recommended that all patients with tinea cruris be subjected to careful examination of all possible mycotic foci, particularly the feet and nails.     

Dermatophytosis due to Trichophyton rubrum. Ten-year period (1996-2006) data collection in a Dermatology Department in Mexico City

Dermatophytosis is the most common mycosis in the world up to 80% caused by Trichophyton rubrum. The aim of the present study was to describe the clinical characteristics of the dermatophytosis caused by T. rubrum in a dermatological outpatient clinic during a ten years period, from 1996 to 2005. We collected the data from patients with a dermatophytosis from which we have isolated T. rubrum. A total of 776 patients with dermatophytosis caused by T. rubrum were found. A slight predominance of female patients (56.2%) was observed. The most commonly affected age group was those in the third to the fifth decade of life, and house working women (33.5%) were predominant. Onychomycosis was found in 63% of the cases, tinea pedis in 22.7%, tinea corporis in 5.2% and tinea cruris in 2.8%. In onychomycosis, the first toe nail was the most commonly affected (58.9%) and the dystrophic type was seen in 50.7% of them.     

亚热带地区部队皮肤真菌病及其病原菌的调查研究

亚热带地区是皮肤真菌病的高发地区,据报道,云南边防某部皮肤真菌病发病率为57.8％,占皮肤病发病率的第一位,美军在越南战争期间,发生的所有皮肤病中,最常见的是皮肤真菌感染,而且造成了非战斗减员。战争后期,美军把皮肤病的防治作为其疾病防治的重点。因此,我们于1992年10月底对海南地区部队皮肤真菌病的发病情况进行了流行病学凋查,并对皮肤真菌病的病原菌进行了分离鉴定,现报告如下。     

Protein profiles of Malassezia pachydermatis isolated from dogs

Chronic dermatophytosis was observed in 2276 (10.02%) of 22 692 patients with dermatophytosis during a period of 5.5 years. Males were affected at least 3 times more frequently than females. The age group most commonly affected was between 20 and 40 years of age. Females were affected more between the ages of 30 to 40 years. Tinea cruris and tinea corporis were the most common clinical types and tinea pedis was the least common type observed. The most frequent isolate was Trichophyton rubrum followed by T. mentagrophytes and T. violaceum. Ichthyosis vulgaris was the most common cutaneous association whereas atopy and diabetes mellitus were the most common systemic associations. This revised version was published online in August 2006 with corrections to the Cover Date.     

2%硝酸舍他康唑乳膏治疗体股癣和足癣疗效和安全性评价

目的 评价2％硝酸舍他康唑乳膏治疗足癣和体、股癣的疗效及安全性,并与2％硝酸咪康唑乳膏对照。方法 采用多中心随机双盲对照试验。试验组及对照组分别外用2％硝酸舍他康唑乳膏和2％硝酸咪康唑乳膏,每天2次,疗程4周。在治疗开始及治疗2周、4周、6周时进行观察。结果 足癣试验组61例,对照组58例;体、股癣试验组56例,对照组55例完成了观察。足癣在4周时,临床有效率试验组98．36％,对照组93．10％,在6周时分别为100％和98．28％;在4周和6周时真菌学清除率试验组均为95．1％,对照组均为100％;在4周时总有效率试验组98．36％,对照组93．10％,在6周时分别为100％和98．28％。体、股癣在4周时,临床有效率试验组98．21％,对照组92．73％,在6周时分别为100％和98．18％;在4周和6周时真菌学清除率试验组和对照组均为100％;在4周时总有效率试验组98．21％,对照组92．73％,在6周时分别为100％和98．18％。药物不良反应发生率试验组1．7％,对照组0．9％,表现为局部刺激。结果 显示2％硝酸舍他康唑乳膏治疗足癣和体、股癣的疗效及安全性与2％硝酸咪康唑乳膏相似（P〉0．05）。结论 2％硝酸舍他康唑乳膏治疗足癣和体、股癣有效、安全。     

A study of 3891 cases of mycoses in the tropics.

4103 cases suspected of mycoses were analysed as to sex, age and site of disease and 3891 were proved cases. This group formed 50% of total mycoses or 13-93% of all dermatoses recorded in the Government General Hospital, Madras, during the period of study. There were 66-26% adult female, 27-6% adult male and 6-14% were below 13 years. Dermatophytoses were found in 73-5%; the other common diseases were tinea versicolor (17-68%) and candidiasis (12-43%). Multiple sites of involvement or more than 1 disease in the same individual were mostly observed. The incidence of piedra (0-1%) and deep mycoses (0-156%) was very low. Mycetoma was the common disease (5/6) in deep mycoses. In dermatophytoses, tinea corporis (49-71%) and tinea cruris (47-85%) commonest; tinea axillaris (3-42%), tinea capitis (1-72%) and tinea barbae (1-29%) were less common. The incidence of tinea manuum, tinea pedis and tinea unguium was similar (4-97%-6-38%). High temperature and humidity were related to the higher incidence of tinea corporis, tinea cruris and tinea versicolor. Mainly children suffered from tinea capitis. All other mycoses were commonly found in adults between 2nd and 3rd decades. In all mycoses but candidiasis, female predominated. Cutaneous candidiasis was mainly a problem of housewives. Among the dermatophytes Trichophyton violaceum was predominant (33-7%) followed by T. rubrum (32-6%). Trichophyton schoenleinii and M. gypseum were rarely isolated. From mycetoma, Madurella mycetomii, Nocardia braziliensis, N. asteroides and Actinomadura spp. were isolated. Demonstration of Cryptococcus laurentii in 1 case is reported in this area for the first time.     

822例浅部真菌病及其致病菌种分析

目的分析新疆乌鲁木齐市浅部真菌病病原菌的流行趋势。方法对2006年1月～2013年12月就诊于我院皮肤科门诊、有典型临床表现、真菌镜检及培养均阳性的822例患者进行致病病种及菌种分析。使用SPSS17．0统计软件对于结果进行统计分析。结果822例患者包括8个病种：体癣214例（26．0％）,足癣131例（15．9％）,头癣128例（15．6％）,甲真菌病125例（15．2％）,股癣119例（14．5％）,手癣54例（6．6％）,皮肤念珠菌病38例（4．6％）,花斑糠疹13例（1．6％）。菌种分布以须癣毛癣菌为首位,336株（40．9％）,其次为红色毛癣菌共239株（29．1％）,犬小孢子菌68株（8．3％）,铁锈色小孢子菌48株（5．8％）,念珠菌38株（4．6％）,断发毛癣菌33株（4．0％）,许兰黄癣菌23株（2．8％）,马拉色菌13株（1．6％）,疣状毛癣菌9株（1．1％）,紫色毛癣菌7株（0．9％）,支顶孢霉菌4株（0．5％）,曲霉、絮状表皮癣菌各2株（各0．2％）。统计学分析显示：不同性别足癣、股癣、甲真菌病及花斑糠疹构成比有统计学意义（P〈0．05）,除体癣及花斑糠疹外,本组其他浅部真菌病的分布在年龄方面有着明显的差别（P〈O．05）。结论乌鲁木齐市浅部真菌病发病率排在前3位的依次为体癣、足癣、头癣,致病病原菌以毛癣菌为主,须癣毛癣菌为首位致病菌,其次为红色毛癣菌。足癣及甲真菌病好发于女性,而股癣及花斑糠疹好发于男性。除体癣及花斑糠疹外,不同年龄段浅部真菌病分布有明显差异,其中头癣主要好发于青少年。     

Trichophyton simii infection in man and animals

Nine human infections due toT. simii comprising of tinea corporis (6), tinea cruris (2) and tinea capitis (1) have been reported. Human cases were having lesions typically of zoophilic contracted infections. Lesions in dogs were on nose and upper lip and were circular. All the strains showed typical and identical macro and microscopie morphology. Three isolates studied by Stockdale were negative (2) and positive (1) strains. One studied here was negative. Possible epidemiology is discussed.