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

摘要/Abstract

摘要：

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，0.032 μg·mL^-1，MIC范围，0.001~0.125 μg·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，0.041 μg·mL^-1，MIC范围，0.032~0.125 μg·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。

关键词: 皮肤癣菌, 病原菌特点, 体外药敏

Abstract:

Objective To analyze the pathogenic characteristics of dermatophyte infection in the out-patient department of a teaching hospital in Shenzhen area and to investigate in vitro activity of 7 antifungal agents against 85 strains of dermatophyte. Methods Skin scales (or nail scraps, hairs) were collected from suspected patients with dermatophyte infection for KOH direct microscopic examination and culture. The species of dermatophyte were identified by sequencing the ribosomal DNA internal transcribed spacer(ITS). The antifungal susceptibility testing was performed following Clinical and Laboratory Standards Institute (CLSI) M38-A2 reference broth microdilution methods. Seven antifungal agents were evaluated against 85 strains of dermatophytes. Results Among 161 suspected patients with dermatophyte infection, the positive rate of KOH direct microscopic examination of dermatophyte was 67.7%, and the positive rate of fungal culture of dermatophyte was 52.8%. A total of 85 isolates of positive fungal culture were identified as 68 strains of Trichophyton rubrum, 7 strains of Microsporum canis, 3 strains of Microsporum gypseum, 5 strains of Trichophyton interdigitale, 1 strain of Trichophyton violaceum and 1 strain of Trichophyton tonsurans. In vitro drug susceptibility test results revealed that terbinafine (GM MIC,0.032μg·mL^-1, MIC range, 0.001~0.125μg·mL^-1) had the strongest in vitro antifungal activity against 6 dermatophytes. And voriconazole (GM MIC,0.041μg·mL^-1, MIC range, 0.032~0.125μg·mL^-1)had the lowest MIC among the azole drugs. Conclusions Trichophyton rubrum was the most common pathogen of dermatophyte infection in our hospital. The standardized antifungal susceptibility testing confirmed that terbinafine and voriconazole had strong in vitro antifungal activity against isolated dermatophyte and were suitable for the treatment of dermatophyte infection in this area.

Key words: dermatophyte, pathogenic characteristics, in vitro susceptibility

中图分类号:

R379.2

刘洁, 刘晓云, 于波, 胡小平. 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[J]. 中国真菌学杂志, 2019, 14(6): 351-356.

LIU Jie, LIU Xiao-yun, YU Bo, HU Xiao-ping. Analysis on pathogenic characteristics of dermatophyte infection and in vitro drug susceptibility of 85 strains of dermatophyte[J]. Chinese Journal of Mycology, 2019, 14(6): 351-356.

参考文献

[1] Lanternier F, Pathan S, Vincent QB, et al. Deep dermatophytosis and inherited CARD9 deficiency[J]. New Engl J Med, 2013, 369(18):1704-1714.
[2] Garcia GH,Hrycyk MF, Giacobino J, et al. Molecular identification and phylogenetical analysis of dermatophyte fungi from Latin America[J]. Mycoses, 2016, 59(12):787-797.
[3] Ellabib MS, Khalifa Z, Kavanagh K, et al. Dermatophytes and other fungi associated with skin mycoses in Tripoli, Libya[J]. Mycoses, 2002, 45(3-4):101-104.
[4] Falahati M, Akhlaghi L, Lari AR, et al. Epidemiology of dermatophytoses in an area south of Tehran, Iran[J]. Mycopathologia, 2003, 156(4):279-287.
[5] Zamani S, Sadeghi G, Yazdinia F, et al. Epidemiological trends of dermatophytosis in Tehran, Iran:A five-year retrospective study[J]. J Mycol Med, 2016, 26(4):351-358.
[6] Muhsin TM, al-Rubaiy KK, al-Duboon AH, et al. Characteristics of dermatophytoses in Basrah, Iraq[J]. Mycoses, 1999, 42(4):335-338.
[7] Hainsworth S, Hamblin JF, Vanniasinkam T, et al. Isolation of dermatophytes (and other fungi) from human nail and skin dust produced by podiatric medical treatments in Australia[J]. J Am Podiatr Med Assoc, 2015, 105(2):111-120.
[8] Muangkaew W, Wongsuk T, Luplertlop N, et al. Common dermatophytes and in vitro anti-fungal susceptibility testing in patients attending the Dermatological Clinic at the Hospital for Tropical Medicine, Bangkok[J]. New Microbiol, 2017, 40(3):175-179.
[9] Silva-Rocha WP, deAzevedo MF, Chaves GM, et al. Epidemiology and fungal species distribution of superficial mycoses in Northeast Brazil[J]. J Mycol Med, 2017, 27(1):57-64.
[10] Vena GA,Chieco P, Posa F, et al. Epidemiology of dermatophytoses:retrospective analysis from 2005 to 2010 and comparison with previous data from 1975[J]. New Microbiol, 2012, 35(2):207-213.
[11] 马志红, 章强强, 鲁长明,等. 不同地区浅部真菌病病原菌流行状况比较[J]. 中国真菌学杂志, 2009, 4(6):343-346.
[12] Antuori A, Fernandez G, Fernandez A, et al. Epidemiology of dermatophytic infections between 2008 and 2017 in Barcelona, Spain[J]. Enferm Infecc Microbiol Clin, 2019. doi:10.1016/j.eimc.2019.02.010.
[13] Cai W, Lu C, Li X, et al. Epidemiology of superficial fungal infections in Guangdong, Southern China:a retrospective study from 2004 to 2014[J]. Mycopathologia, 2016, 181(5-6):387-395.
[14] Shehata AS, Mukherjee PK, Aboulatta HN, et al. Single-step PCR using (GACA)₄ primer:utility for rapid identification of dermatophyte species and strains[J]. J Clin Microbiol, 2008,46(8):2641-2645.
[15] Dhib I, Fathallah A, Yaacoub A, et al. Multiplex PCR assay for the detection of common dermatophyte nail infections[J]. Mycoses, 2014, 57(1):19-26.
[16] Liu D,Coloe S, Baird R, et al. Molecular determination of dermatophyte fungi using the arbitrarily primed polymerase chain reaction[J]. Br J Dermatol, 1997, 137(3):351-355.
[17] Verrier J, Krahenbuhl L, Bontems O, et al. Dermatophyte identification in skin and hair samples using a simple and reliable nested polymerase chain reaction assay[J]. Br J Dermatol, 2013, 168(2):295-301.
[18] Kanbe T, Suzuki Y, Kamiya A, et al. Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes[J]. J Dermatol Sci, 2003, 33(1):41-54.
[19] Hryncewicz-Gwozdz A, Jagielski T, Dobrowolska A, et al. Identification and differentiation of Trichophyton rubrum clinical isolates using PCR-RFLP and RAPD methods[J]. Eur J Clin Microbiol Infect Dis, 2011, 30(6):727-731.
[20] Nenoff P, Erhard M, Simon JC, et al. MALDI-TOF mass spectrometry-a rapid method for the identification of dermatophyte species[J]. Med Mycol, 2013,51(1):17-24.
[21] deRespinis S, Tonolla M, Pranghofer S, et al. Identification of dermatophytes by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. Med Mycol, 2013, 51(5):514-521.
[22] 吴伟伟, 王千, 龚杰, 等. 8种抗真菌药物对浅表真菌病致病菌株体外敏感性检测[J]. 中国真菌学杂志, 2017, 12(5):279-82, 278.
[23] 卜晓琳, 范娟, 胡晓莉, 等. CLSI M38-A2及M27-A3方案分析足癣36株真菌的药敏特征[J]. 中国真菌学杂志, 2016, 11(3):169-173.
[24] Silva LB, de Oliveira DB, Da SB et al. Identification and antifungal susceptibility of fungi isolated from dermatomycoses[J]. JEur Acad Dermatol Venereol, 2014, 28(5):633-640.
[25] Altinbas R, Ozakkas F, Baris A, et al. In vitro susceptibility of seven antifungal agents against dermatophytes isolated in Istanbul[J]. Turk J Med Sci, 2018, 48(3):615-619.
[26] Digby SS,Hald M, Arendrup MC, et al. Darier disease complicated by terbinafine-resistant Trichophyton rubrum:a case report[J]. Acta Derm Venereol, 2017, 97(1):139-140.
[27] Salehi Z, Shams-Ghahfarokhi M, Razzaghi-Abyaneh M, et al. Antifungal drug susceptibility profile of clinically important dermatophytes and determination of point mutations in terbinafine-resistant isolates[J]. Eur J Clin Microbiol Infect Dis, 2018, 37(10):1841-1846.
[28] Kano R, Hsiao YH, Han HS, et al. Resistance mechanism in aterbinafine-resistant strain of Microsporum canis[J]. Mycopathologia, 2018, 183(3):623-627.
[29] Hiruma J, Kitagawa H, Noguchi H, et al. Terbinafine-resistant strain of Trichophyton interdigitale strain isolated from a tinea pedis patient[J]. J Dermatol, 2019,46(4):351-353.
[30] Deng S, Zhang C, Seyedmousavi S, et al. Comparison of the in vitro activities of newer triazoles and established antifungal agents against Trichophyton rubrum[J]. Antimicrob Agents Chemother, 2015, 59(7):4312-4314.
[31] 颜文良, 胡文星, 刘芳, 等. 伏立康唑首次成功治疗难治性面部Majocchi肉芽肿1例并文献复习[J].中国真菌学杂志, 2016, 11(5):265-268.