中国南北方地区临床孢子丝菌菌种鉴定

中国真菌学杂志　 2015, Vol. 10 　Issue (5): 279-282.

中国南北方地区临床孢子丝菌菌种鉴定

贺羽¹, 黄梦雅¹, 胡青碧¹, 周汛^1,2

1. 重庆医科大学附属第一医院皮肤科, 重庆 400016;
2. 重庆市中医院皮肤美容科, 重庆 400021

收稿日期:2015-06-23 出版日期:2015-10-28 发布日期:2015-10-28
通讯作者: 周汛,E-mail:zhouxun123@sina.com E-mail:zhouxun123@sina.com
作者简介:贺羽,女(汉族),硕士研究生在读.E-mail:15123825300@163.com
基金资助:
国家自然科学基金(31270062)

Species identification of Sporothrix clinical isolates in Northern and Southern of China

HE Yu¹, Huang Mengya¹, HU Qing-bi¹, ZHOU Xun^1,2

1. Department of Dermatology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;
2. Department of Dermatology and Cosmetology, Chongqing Hospital of Traditional Chinese Medicine, Chongqing 400021, China

Received:2015-06-23 Online:2015-10-28 Published:2015-10-28

摘要/Abstract

摘要：

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。

关键词: 孢子丝菌, 形态学, 系统发育, 球形孢子丝菌

Abstract:

Objective To identify 50Sporothrix clinical isolates in Northern and Southern of China.Methods All isolates from patients were subcultured at 25℃ and morphological characteristic was analysed by microscope and visual inspection.The total genomic DNA was extracted to amplify the partial β-tubulin gene and the internal transcribed(ITS) region by polymerase chain reaction(PCR).The phylogenetic tree was constructed by DNA sequences analyses based on datasets of ITS and a combined ITS and partial β-tubulin region using maxium likehood(ML) and neighbor-joining(NJ) methods with MEGA5 software.The Reference sequences used for phylogenetic analyses were retrieved from GenBank.Result The macroscopic morphologies and microscopic features of all isolates were comformed to Sporothrix spp.The phylogenetic tree showed that all of the isolates were clustered in a distinct clade with a type of Sporothrix globosa.Conclusion S.globosa may be the major Sporothrix existing in northern and southern of China, which can provide basis for the epidemiological investigation of Sporothrix.

Key words: Sporothrix, morphology, phylogeny, Sporothrix globosa

中图分类号:

R519.8

贺羽, 黄梦雅, 胡青碧, 周汛. 中国南北方地区临床孢子丝菌菌种鉴定[J]. 中国真菌学杂志, 2015, 10(5): 279-282.

HE Yu, Huang Mengya, HU Qing-bi, ZHOU Xun. Species identification of Sporothrix clinical isolates in Northern and Southern of China[J]. Chinese Journal of Mycology, 2015, 10(5): 279-282.

参考文献

[1] Mahajan VK.Sporotrichosis:an overview and therapeutic options[J].Dermatol Res Pract,2014,2014:272376.
[2] Marimon R,Gene J,Cano J,et al.Molecular phylogeny of Sporothrix schenckii[J].J Clin Microbiol,2006,44(9):3251-3256.
[3] Tellez MD,Batista-Duharte A,Portuondo D,et al.Sporothrix schenckii complex biology:environment and fungal pathogenicity[J].Microbiology,2014,160(Pt 11):2352-2365.
[4] Moreira JA,Freitas DF,Lamas CC.The impact of sporotrichosis in HIV-infected patients:a systematic review[J].Infection,2015,43(3):267-276.
[5] Fernandes GF,Dos SP,Rodrigues AM,et al.Characterization of virulence profile,protein secretion and immunogenicity of different Sporothrix schenckii sensu stricto isolates compared with S.globosa and S.brasiliensis species[J].Virulence,2013,4(3):241-249.
[6] Song Y,Li SS,Zhong SX,et al.Report of 457 sporotrichosis cases from Jilin province,northeast China,a serious endemic region[J].J Eur Acad Dermatol Venereol,2013,27(3):313-318.
[7] Liu TT,Zhang K,Zhou X.Molecular identification of Sporothrix clinical isolates in China[J].J Zhejiang Univ Sci B,2014,15(1):100-108.
[8] Yu X,Wan Z,Zhang Z,et al.Phenotypic and molecular identification of Sporothrix isolates of clinical origin in Northeast China[J].Mycopathologia,2013,176(1-2):67-74.
[9] 黄怀球,袁立燕,张静,等.申克孢子丝菌的扫描电镜观察[J].热带医学杂志,2012,(12):1442-1443.
[10] Allen GC,Flores-Vergara MA,Krasynanski S,et al.A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide[J].Nat Protoc,2006,1(5):2320-2325.
[11] Zharkikh A,Li WH.Estimation of confidence in phylogeny:the complete-and-partial bootstrap technique[J].Mol Phylogenet Evol,1995,4(1):44-63.
[12] Lopes-Bezerra LM,Schubach A,Costa RO.Sporothrix schenckii and sporotrichosis[J].An Acad Bras Cienc,2006,78(2):293-308.
[13] Madrid H,Cano J,Gene J,et al.Sporothrix globosa,a pathogenic fungus with widespread geographical distribution[J].Rev Iberoam Micol,2009,26(3):218-222.
[14] Carlos IZ,Sassa MF,Da GSD,et al.Current research on the immune response to experimental sporotrichosis[J].Mycopathologia,2009,168(1):1-10.
[15] Arrillaga-Moncrieff I,Capilla J,Mayayo E,et al.Different virulence levels of the species of Sporothrix in a murine model[J].Clin Microbiol Infect,2009,15(7):651-655.
[16] Almeida-Paes R,de Oliveira LC,Oliveira MM,et al.Phenotypic characteristics associated with virulence of clinical isolates from the Sporothrix complex[J].Biomed Res Int,2015,2015:212308.
[17] Marimon R,Cano J,Gene J,et al.Sporothrix brasiliensis,S.globosa,and S.mexicana,three new Sporothrix species of clinical interest[J].J Clin Microbiol,2007,45(10):3198-3206.
[18] Marimon R,Serena C,Gene J,et al.In vitro antifungal susceptibilities of five species of Sporothrix[J].Antimicrob Agents Chemother,2008,52(2):732-734.
[19] Ottonelli SC,Magagnin CM,Castrillon MR,et al.Antifungal susceptibilities and identification of species of the Sporothrix schenckii complex isolated in Brazil[J].Med Mycol,2014,52(1):56-64.
[20] Oliveira MM,Almeida-Paes R,Muniz MM,et al.Phenotypic and molecular identification of Sporothrix isolates from an epidemic area of sporotrichosis in Brazil[J].Mycopathologia,2011,172(4):257-267.
[21] 谭静文,刘伟,万喆,等.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[J].菌物学报,2013(02):162-168.
[22] Oliveira MM,Santos C,Sampaio P,et al.Development and optimization of a new MALDI-TOF protocol for identification of the Sporothrix species complex[J].Res Microbiol,2015,166(2):102-110.
[23] Camacho E,Leon-Navarro I,Rodriguez-Brito S,et al.Molecular epidemiology of human sporotrichosis in Venezuela reveals high frequency of Sporothrix globosa[J].BMC Infect Dis,2015,15:94.