福建地区94例女性阴道念珠菌感染分离株分型研究

摘要/Abstract

摘要： 目的对福建地区临床阴道分泌物分离出的（94株）念珠菌菌株进行分类鉴定，并通过分子生物学及API试验分析传统科玛嘉分类方法的准确性。方法 1通过科玛嘉显色培养基鉴定菌种，并观察菌株显微镜下形态学表现。2通过ITS区段分子生物学序列分析进行菌种鉴定。3将科玛嘉试验与ITS区段序列分析结果与化验室检验报告结果对比，将鉴定差别菌株进一步行API试验及LSU区段序列分析。结果 194株念珠菌经鉴定结果为白念珠菌78株、光滑念珠菌10株、近平滑念珠菌3株、热带念珠菌1株、酿酒假丝酵母菌1株及其中1株为光滑念珠菌及近平滑念珠菌混合感染。2科玛嘉试验可良好的鉴定念珠菌，但对于少见菌种（如酿酒假丝酵母菌）仍缺乏特异性。3一般化验室通过简单菌落形态学及简易科玛嘉检测鉴定仍存在一定误差率（10/94）。4分子生物学方法鉴定菌种准确性高，且可从基因序列分析中鉴定包含C.parapsilosis sensu strico，Candida metapsilosis以及Candida orthopsilosis的近平滑念珠菌复合体菌种。结论福建地区女性外阴感染的菌种仍以白念珠菌为主，但非白念珠菌的感染也占据相当的比例（16/94），而在检测方法上，分子生物学技术较科玛嘉试验更能准确的鉴定念珠菌菌种。

关键词: 念珠菌, 鉴定, 分子生物学, 科玛嘉培养基

Abstract: Objective To reclassify the 94 Candida strains from clinical vulvovaginal Candidiasis patients in Fujian area, and to evaluate the accuracy of CHROM agar identification by comparing with the API test and molecular biological analysis.Methods 1Identify the 94 strains by CHROM agar, and observe the microscopic morphology. 2Identify these strains by ITS sequence analysis of molecular biological method. 3Comparing the test reports from the clinical laboratory with the identification results of CHROM agar and ITS sequence analysis, the isolates in doubt are selected to have the API test and LSU sequence analysis.Results 1There were 78 strains identified as C.albicans, 10 strains as C.glabrata, 3 strains as C.parasilosis, 1 strain as Saccharomyces cerevisiae, and 1 strains as both C.glabrata and C.parasilosis. 2The CHROM agar could nearly correctly identify Candida, however, lacks specificity when it comes to rare isolates (ex.Saccharomyces cerevisiae). 3According to the comparing results, we could conclude that the identification of clinical laboratory just by the methods of simple morphology and CHROM agar test still has the error rate (10/94). 4The identification of molecular biological method had high accuracy rate. Besides, we could identify the C.parapsilosis complex was consist of the species of C.parapsilosis sensu strico,Candida metapsilosis and Candida orthopsilosis by this way.Conclusions The most part of clinical isolates of vulvovaginal candidiasis in Fujian area were still C.albicans, but the non C.albicans also occupied a certain proportion (16/94). Our result showed that the molecular biological methods were more accurate than the CHROM agar test in identification.

Key words: candidiasis, identification, molecular biology, CHROM agar

中图分类号:

R519.3

陈柏叡, 江爱萍, 徐明, 卢万丁, 蔡蕾. 福建地区94例女性阴道念珠菌感染分离株分型研究[J]. 中国真菌学杂志, 2014, 9(4): 222-227.

CHEN Bo-rui, JIANG Ai-ping, XU Ming, LU Wan-ding, CAI Lei. The identification of 94 clinical isolates from vulvovaginal candidiasis in Fujian area[J]. Chinese Journal of Mycology, 2014, 9(4): 222-227.

参考文献

[1] Latouche GN, Daniel HM, Lee OC, et al. Comparison of use of phenotypic and genotypic characteristics for identification of species of the anamorph genus Candida and related teleomorph yeast species[J]. J Clin Microbiol, 1997, 35(12):3171-3180.
[2] Sullivan DJ, Henman MC, Moran GP,et al. Molecular genetic approaches to identification, epidemiology and taxonomy of non-albicans Candida species[J]. J Med Microbiol, 1996, 44(6):399-408.
[3] Ciardo DE, Schar G, Bottger EC,et al. Internal transcribed spacer sequencing versus biochemical profiling for identification of medically important yeasts[J]. J Clin Microbiol, 2006, 44(1):77-84.
[4] Nilsson RH, Kristiansson E, Ryberg M, et al. Intraspecific ITS variability in the kingdom fungi as expressed in the international sequence databases and its implications for molecular species identification[J]. Evol Bioinform Online, 2008, 4(26):193-201.
[5] Lachance MA, Daniel HM, Meyer W, et al. The D1/D2 domain of the large-subunit rDNA of the yeast species Clavispora lusitaniae is unusually polymorphic[J]. FEMS Yeast Res,2003, 4(3):253-258.
[6] Tavanti A, Davidson AD, Gow NA, et al.Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III[J]. J Clin Microbiol,2005, 43(1):284-292.
[7] Vaughan-Martini A, Kurtzman CP, Meyer SA, et al. Two new species in the Pichia guilliermondii clade:Pichia caribbica sp. nov., the ascosporic state of Candida fermentati, and Candida carpophila comb. nov[J]. FEMS Yeast Res ,2005, 5(4-5):463-469.
[8] Vermitsky JP, Self MJ, Chadwick SG, et al.Survey of vaginal-flora Candida species isolates from women of different age groups by use of species-specific PCR detection[J]. J Clin Microbiol ,2008, 46(4):1501-1503.
[9] Richter SS, Galask RP, Messer SA, et al. Antifungal susceptibilities of Candida species causing vulvovaginitis and epidemiology of recurrent cases.[J] J Clin Microbiol ,2005, 43(5):2155-2162.
[10] Spinillo A, Capuzzo E, Gulminetti R, et al. Prevalence of and risk factors for fungal vaginitis caused by non-albicans species[J]. Am J Obstet Gynecol,1997, 176(1 Pt 1):138-141.
[11] Almirante B, Rodriguez D, Park BJ, et al. Epidemiology and predictors of mortality in cases of Candida bloodstream infection: results from population-based surveillance, barcelona, Spain, from 2002 to 2003[J]. J Clin Microbiol ,2005, 43(4):1829-1835.
[12] Miranda LN, van der Heijden IM, et al.Candida colonisation as a source for candidaemia[J]. J Hosp Infect ,2009, 72(1):9-16.
[13] Nakamura T, Takahashi H: Epidemiological study of Candida infections in blood: susceptibilities of Candida spp. to antifungal agents, and clinical features associated with the candidemia[J]. J Infect Chemother ,2006, 12(3):132-138.
[14] Pfaller MA, Diekema DJ, Gibbs DL, et al.Geographic and temporal trends in isolation and antifungal susceptibility of Candida parapsilosis:a global assessment from the ARTEMIS DISK Antifungal Surveillance Program, 2001 to 2005[J]. J Clin Microbiol, 2008, 46(3):842-849.
[15] Chen YC,Lin YH,Chen KW,et al.Molecular epidemiology and antifungal susceptibility of Candida parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis in Taiwan. [J].Diagn Microbiol Infect Dis.2010, 68(3): p. 284-292.
[16] Orsi CF, Colombari B, Blasi E.Candida metapsilosis as the least virulent member of the 'C.parapsilosis' complex[J]. Med Mycol, 48(8):1024-1033.