中国南北方地区临床孢子丝菌菌种鉴定

目的对分离自我国南、北方地区50株临床孢子丝菌进行菌种鉴定。方法分离菌株分别进行25℃恒温培养和玻片小培养,肉眼和镜下观察形态特征;同时提取菌丝相基因组DNA,用PCR分别扩增部分微管蛋白(β-tubulin)基因和核糖体内部转录间隔区(Ribosomal Internal Transcribed Spacer,ITS),扩增产物进行测序,并采用最大似然法(maximum likelihood,ML)和邻接法(neighbor-joining,NJ)构建联合β-tubulin基因和ITS区域的系统发育树。结果结合形态学及系统发育分析,50株菌均鉴定为球形孢子丝菌(Sporothrix globosa,S.globosa)。结论球形孢子丝菌是目前我国南、北方地区孢子丝菌病的主要致病菌种,为进一步明确我国孢子丝菌病致病菌种的分布提供了依据。     

1株致体股癣的颗粒型红色毛癣菌变种的形态学观察与基因鉴定

目的：探讨1株致体股癣的颗粒型红色毛癣菌变种的形态学特点及核糖体基因序列变化。方法1例体股癣患者皮损中分离出1株红色毛癣菌,观察形态学特点,并用PCR测定核糖体转录间区（ITS）、D1/D2区、非转录区基因序列（NTS区）内串联重复亚单位（TRS）。结果该菌株可产生鹿角形菌丝,大分生孢子有中空现象及单侧出芽产孢特征,可在42℃生长;ITS和D1/D2区基因测序鉴定为红色毛癣菌,但TRS-1和TRS-2基因序列分别有2个、1个碱基差异。结论该菌株可能是颗粒型红色毛癣菌的1个变种。     

申克孢子丝菌基因分型的研究现状与展望

申克孢子丝菌以往被认为单一菌种,呈全球性分布。然而不同区域来源的菌株,在药物敏感性、致病力等方面差异较大。RAPD、RFLP等多种分子生物学技术曾被用于该菌基因分型的探讨。近年来,以CAL基因结合菌株表型的分类方法,已成为申克孢子丝菌复合体公认的划分方法。本文将对申克孢子丝菌的基因分型方法进行综述,并着重阐述CAL基因分型的研究进展。     

孢子丝菌相关复合种需加强生物安全检控

近年来,随着分子生物学及真菌分类研究的快速发展,包括检疫性真菌在内的物种分类及命名已发生翻天覆地的变化。孢子丝菌病的病原菌长期被认为有且仅有申克孢子丝菌1种,现经多基因系统发育学分析发现孢子丝菌病的病原菌并非仅为申克孢子丝菌,而由6个种构成。我国孢子丝菌菌株的分子生物学鉴定,目前只发现了球形孢子丝菌和申克孢子丝菌,其他4种孢子丝菌尚未发现传入我国。由于这些物种之间形态差异小,但致病力、致病反应、药物反应各不形同,应加强生物安全管控和监测,防止这些有害真菌传入我国。     

免疫抑制沙鼠源肺孢子菌的分类地位

肺孢子菌能够感染多种哺乳动物的肺脏, 并在免疫受抑制的个体引起肺孢子菌肺炎. 研究表明, 肺孢子菌分离株在系统进化树上分为两大枝, 二者所含菌株分别来源于灵长类和啮齿类. 基于宿主特异性以及DNA序列和形态学差异, 每一分枝内的菌株又可分为不同的种. 本研究采用地塞米松免疫抑制法, 导致沙鼠(28/35)肺组织感染. 对18S, 5.8S rRNA和rRNA内转录间隔区基因序列, 以及线粒体核糖体大亚基基因的部分序列分析表明, 本实验沙鼠源肺孢子菌与啮齿类动物来源的同属一枝, 但明显区别于其他啮齿类动物来源的菌株, 应考虑是一个独立的菌种.     

孢子丝菌复合体分子分型研究进展

孢子丝菌复合体属于双相真菌,全球分布,可引起人类及动物的慢性深部感染。不同地域的菌株在致病力、传播途径及药物敏感性等方面均存在差异。孢子丝菌病作为一种人兽共患病,其发病率逐年上升,出现多次暴发流行。分子分型不仅是明确感染源和传播途径、预防和控制疾病流行的有力手段,同时有助于了解孢子丝菌基因型与表型的相关性,在研究其致病机制以及临床诊治过程中都具有十分重要的意义。本文对孢子丝菌的分子分型方法的研究进行综述。     

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。     

来自中国北方部分地区33株孢子丝菌菌株的表型和基因型鉴定

孢子丝菌病是一种临床常见的真菌感染疾病,近期有研究认为其致病菌是由多个同形种构成的复合体。为明确我国孢子丝菌病致病菌的生理学以及分子生物学方面的特点,对33株分离自我国孢子丝菌病患者组织的病原菌进行了研究,首先检测其在37℃是否生长以及PDA培养基上生长21d的菌落直径,其次检测其糖同化特点,最后对其钙调蛋白(Calmodulin,CAL)基因进行PCR扩增、序列测定以及系统进化分析。结果显示,这些菌株全部为球形孢子丝菌Sporothrix globosa,说明我国孢子丝菌病的病原菌可能以球形孢子丝菌为主。     

1例皮肤播散型孢子丝菌病的分离菌株的基因鉴定

孢子丝菌病是由申克孢子丝菌及其卢里变种感染引起的皮肤、皮下组织和附近淋巴系统的亚急性和慢性感染,偶可播散至全身引起多系统损害。该病遍布全世界,我国孢子丝菌病主要是由申克孢子丝菌引起。孢子丝菌一般生存在土壤和植物上,人的皮肤接触带菌植物或土壤后可引起感染。临床上主要表现为固定型和淋巴管型孢子丝菌病,皮肤播散型孢子丝菌病少见。现认为机体感染申克孢子丝菌引起的不同的临床型别与机体免疫状态、申克孢子丝菌的致病力及其基因差异相关[1-2]。我     

实时荧光PCR检测巴西孢子丝菌的实验方法的建立

目的建立一种快速、特异、灵敏的荧光PCR方法检测巴西孢子丝菌。方法比对NCBI数据库中所有巴西孢子丝菌内转录间隔区(internal transcribed spacer, ITS)序列,在保守区域设计并合成特异性引物和探针,建立并优化荧光PCR检测方法。对优化后的方法使用标准浓度核酸进行扩增效率、灵敏度及特异度评价。通过巴西孢子丝菌小鼠感染模型,与组织培养比较,对本研究中的方法进行评价。结果建立的实时荧光PCR方法对巴西孢子丝菌的检测限为100fg。该方法对申克孢子丝菌、球形孢子丝菌、其他常见致病真菌28种、常见细菌3种以及人类基因组和小鼠基因组扩增结果均为阴性,特异度为100%。对巴西孢子丝菌感染小鼠脑、肝、肺、脾、肾及淋巴结检测与培养结果相一致。结论本研究建立的荧光PCR方法可快速、灵敏、特异地鉴定巴西孢子丝菌,并能够有效的对感染小鼠模型标本进行检测,有助于孢子丝菌病的早期特异性病原学诊断。     

Variation in Genotype and Higher Virulence of a Strain of <Emphasis Type="Italic">Sporothrix schenckii</Emphasis> Causing Disseminated Cutaneous Sporotrichosis

Sporotrichosis is usually a localized, lymphocutaneous disease, but its disseminated type was rarely reported. The main objective of this study was to identify specific DNA sequence variation and virulence of a strain of Sporothrix schenckii isolated from the lesion of disseminated cutaneous sporotrichosis. We confirmed this strain to be S. schenckii by^® tubulin and chitin synthase gene sequence analysis in addition to the routine mycological and partial ITS and NTS sequencing. We found a 10-bp deletion in the ribosomal NTS region of this strain, in reference to the sequence of control strains isolated from fixed cutaneous sporotrichosis. After inoculated into immunosuppressed mice, this strain caused more extensive system involvement and showed stronger virulence than the control strain isolated from a fixed cutaneous sporotrichosis. Our study thus suggests that different clinical manifestation of sporotrichosis may be associated with variation in genotype and virulence of the strain, independent of effects due to the immune status of the host.     

伊曲康唑联合特比萘芬治疗孢子丝菌病疗效及其对病原真菌体外抗菌活性研究

目的 初步探讨伊曲康唑和特比萘芬联合治疗孢子丝菌病的疗效,评价两药体外联合对申克孢子丝菌菌丝相和酵母相的抗菌活性.方法 口服伊曲康唑200mg/d和特比萘芬250mg/d治疗孢子丝菌病;体外联合药敏试验采用棋盘微量稀释法,计算分数抑菌浓度(FIC)指数判定两药相互作用具有协同、拮抗或无关作用.结果 伊曲康唑和特比萘芬联...     

鼻部固定型孢子丝菌病1例

报道1例猫抓后引起的固定型孢子丝菌病.患者男,16岁,皮损表现为鼻翼部位的增生物,其上覆有脓痂.临床上易与细菌感染混淆,但根据患者的病史、临床表现、病理、真菌镜检及培养诊断为申克孢子丝菌引起的孢子丝菌病.患者在应用7个月的碘化钾结合特比萘芬软膏外用治疗后,皮损完全消失.     

Sporotrichosis in patient with AIDS: report of a case and review

Although sporotrichosis is not an AIDS-defining infection, reports of sporotrichosis in individuals infected with HIV are increasing. We report an unusual case of this co-infection in a man with progressive deep cutaneous ulcerations with numerous pleomorphic yeast cells of Sporothrix schenckii. In addition a review of the literature on this subject was carried out and commented upon.     

Isolation of Sporothrix schenckii from environmental samples in Venezuela

The dimorphic fungus Sporothrix schenckii is the etiological agent of sporotrichosis, a subcutaneous mycosis frequently found in Latin America. The isolation of this fungus from the environment and other sources has been widely reported. Nevertheless, to our knowledge this fungus has not been isolated from the endemic areas of Venezuela. In studies related to a clinical case of sporotrichosis in "Colonia Tovar", produced by traumatism after manipulating soil samples, the fungus was isolated from the soil of that particular area. This is the first report of the isolation of S. schenckii from environmental sources in an endemic area of Venezuela.     

Humoral immune response against soluble and fractionate antigens in experimental sporotrichosis

Sporotrichosis is a chronic granulomatous mycosis caused by the dimorphic fungus Sporothrix schenckii, which is widely distributed in nature, and presents a saprophytic mycelial form on plant debris and soil. The immunological mechanisms involved in the prevention and control of sporotrichosis are not yet fully understood. In this study, mice were studied after infection with Sporothrix schenckii. In the first week after infection, fungal loading increased and thence decreased drastically 14 days after infection. Analysis by immunoblotting showed that the sera of all mice tested had antibodies reacting only with a 70 kDa antigen, with predominance of IgG1 and IgG3. Taken together, our results show that antigens from S. schenckii induced a specific humoral response in infected mice.     

Effect of potassium iodide on the immune response in the sporotrichosis.

Potassium iodide (KI), the specific treatment for sporotrichosis, apparently does not have a direct action on Sporothrix schenckii. The spontaneous healing and the variability of the clinical presentation in the disease have strengthened the idea that the KI rather interacts with the immune response of the host. The phagocytic process is inefficient in individuals with sporotrichosis in whom the microbicidal mechanism of halogenation fails to control the disease. There is evidence that blocking of free radicals decreases in the presence of KI. Humoral and cellular immunity are present in sporotrichosis but its participation is uncertain; it is yet to be determined if in this mycosis the KI influences other processes or factors of immune response.     

Evaluation of the in vitro and in vivo dimorphism of Sporothrix schenckii, Blastomyces dermatitidis, and Paracoccidioides brasiliensis isolates after preservation in mineral oil

Morphological differentiation has commanded attention for its putative impact on the pathogenesis of invasive fungal infections. We evaluated in vitro and in vivo the dimorphism from mycelial to yeast-phase of Sporothrix schenckii, Blastomyces dermatitidis and Paracoccidioides brasiliensis isolates, two strains for each species, preserved in mineral oil. S. schenckii strains showed typical micromorphology at 25 degrees C but one strain was unable to complete the dimorphic process in vitro. After in vivo passage through mice the strains had the ability to turn into yeast-like cells and to form colonies on brain-heart infusion medium at 36 degrees C. B. dermatitidis strains grew as dirty white to brownish membranous colonies at 25 degrees C and their micromorphology showed thin filaments with single hyaline conidia. At 36 degrees C the colonies did not differ from those grown at 25 degrees C, but produced a transitional micromorphology. P. brasiliensis strains grew as cream-colored cerebriform colonies at 25 degrees C showing a transitional morphology. B. dermatitidis and P. brasiliensis strains did not turn into yeast-like cells in vivo. The present results demonstrate that B. dermatitidis and P. brasiliensis strains were unable to complete the dimorphic process even after in vivo passage, in contrast to the S. schenckii strain.     

Lymphocutaneous Sporotrichosis in an Immunocompetent Patient: A Case Report from Extreme Southern Italy

Sporotrichosis is a mycotic infection rarely encountered in European countries, including Italy. We describe lymphocutaneous sporotrichosis in an immunocompetent 64-year-old male Italian mason. The patient had linear nodules along the right upper arm with severe lymphadenopathy in the same arm. Sporothrix schenckii was identified by phenotypic and molecular methods. Treatment with itraconazole and methylprednisolone followed by administration of itraconazole alone successfully cured the infection. To our knowledge, this is the first documented report of sporotrichosis from Calabria and Sicily, extreme southern Italy.