应用基质辅助激光电离飞行时间质谱快速鉴定临床酵母菌

中国真菌学杂志　 2012, Vol. 　Issue (5): 269-272.

应用基质辅助激光电离飞行时间质谱快速鉴定临床酵母菌

靳颖¹, 杨爽风², 王俊妨¹, 刘锦¹, 崔云涛¹, 周鑫¹, 赵化冰²

1. 武警后勤学院附属医院检验科, 天津 300162;
2. 武警后勤学院职业与环境危害生物标志物重点实验室, 天津 300162

收稿日期:2012-05-31 出版日期:2012-10-28 发布日期:2012-10-28
通讯作者: 赵化冰,E-mail:huabingzhao@yahoo.com.cn E-mail:huabingzhao@yahoo.com.cn
作者简介:靳颖,女(汉族),硕士,副主任技师.E-mail:jinying9032@hotmail.com

Rapid clinical yeast identification by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

JIN Ying¹, YANG Shuang-feng², WANG Jun-fang¹, LIU Jin¹, CUI Yun-tao¹, ZHOU Xin¹, ZHAO Hua-bing²

Department of laboratory, The Affiliated Hospital of Logistical College of Chinese People's Armed Police Force, Tianjin 300162, China

Received:2012-05-31 Online:2012-10-28 Published:2012-10-28

摘要/Abstract

摘要： 目的研究基质辅助激光解析电离飞行时间质谱 (Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry,MALDI-TOF-MS)用于快速检测鉴定临床分离的酵母菌的可行性。方法应用Bruker MALDI-TOF-MS和VITEK 2-compact系统分别鉴定150株临床分离的酵母菌,结果不一致的菌株通过基因序列测定来鉴定。结果 MALDI-TOF-MS快速准确鉴定出了150株临床酵母菌,鉴定符合率在属水平上为100%,种水平上为94%。结论基于MALDI-TOF-MS鉴定方法具有很好的可重复性和准确性,并且其检测成本较低,实验准备时间很短,MALDI-TOF-MS可以用于临床分离的酵母菌的快速鉴定。

关键词: 基质辅助激光解析电离飞行时间质谱, 临床酵母菌, 快速鉴定

Abstract: Objective To establish a rapid identification method of clinical yeast.Methods Total 150 isolated clinical yeast strains were analyzed by Bruker Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS)and VITEK 2-compact system respectively, and the different results were determined by gene sequencing.Results All 150 clinical yeast strains were identified accurately and rapidly by MALDI-TOF-MS. A total of 100% of isolates were identified correctly at the genus level, with 94% identified correctly at the species level.Conclusion The MALDI-TOF MS-based method was found to be reproducible and accurate, with low consumable costs and minimal preparative time. These results demonstrated that MALDI-TOF-MS could be a rapid technique for identification of clinical yeast.

Key words: MALDI-TOF-MS, clinical yeast, rapid identification

中图分类号:

R446.53

靳颖, 杨爽风, 王俊妨, 刘锦, 崔云涛, 周鑫, 赵化冰. 应用基质辅助激光电离飞行时间质谱快速鉴定临床酵母菌[J]. 中国真菌学杂志, 2012, (5): 269-272.

JIN Ying, YANG Shuang-feng, WANG Jun-fang, LIU Jin, CUI Yun-tao, ZHOU Xin, ZHAO Hua-bing. Rapid clinical yeast identification by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry[J]. Chinese Journal of Mycology, 2012, (5): 269-272.

参考文献

[1] Chen S,Marriott D,Nguyen Q,et al. Candidaemia with uncommon Candida species: predisposing factors, outcome,antifungal susceptibility and implications for management[J]. Clin Microbiol Infect, 2009,15(7):662-669.
[2] 孙宗科,张伟,陈西平.应用飞行时间质谱仪快速鉴定细菌的初步研究[J].卫生研究,2004,33(5):552-554.
[3] Chang HC,Leaw SN,Huang AH,et al. Rapid Identification of Yeasts in Positive Blood Cultures by a Multiplex PCR Method[J].J Clinical Microbiology,2001,39(10):3466-3471.
[4] Pfaller M,Diekema D J. Epidemiology of invasive candidiasis: a persistent public health problem[J]. Clin Microbiol Rev, 2007,20(1):133-163.
[5] Montero C I, Shea Y, Jones P, Harrington M, et al. Evaluation of pyrosequencing (R) technology for the identification of clinically relevant non-dematiaceous yeasts and related species[J]. Clin Microbiol Infect, 2008, 27(9):821-830.
[6] Putignani L, Paglia M, Bordi E, et al.Identification of clinically relevant yeast species by DNA sequence analysis of the D2 variable region of the 25-28S rRNA gene[J]. Mycoses,2008,51(3):209-227.
[7] Degand N, Carbonnelle E, Dauphin B, Beretti J,et al. Matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of nonfermenting gram-negative bacilli isolated from cystic fibrosis patients[J]. J Clin Microbiol, 2008,46(10):3361-3367.
[8] Qian J, Cutler J E, Cole R B, and Cai Y. MALDI-TOF mass signatures for differentiation of yeast species, strain grouping and monitoring of morphogenesis markers[J]. Anal Bioanal Chem, 2008,392(3):439-449.
[9] Valentine N, Wunschel K, Wunschel D, Petersen C, Wahl K. Effect of culture conditions on microorganism identification by matrix-assisted laser desorption ionization mass spectrometry[J]. Appl Environ Microbiol,2005,71(1):58-64.
[10] Fenselau C,Demirev PA.Characterizati0n of intact microorganisms by MALDI mass spectrometry[J].Mass Spectrometry Reviews,2001,20(4):157-171.
[11] Stevenson LG, Drake SK,Shea YR,et al. Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of clinically important yeast species [J]. J Clin Microbiol,2010,48(10):3482-3486.
[12] Marklein G, Josten M, Klanke U, Muller E, et al. Matrix-assisted laser desorption ionization time of flight mass spectrometry for fast and reliable identification of clinical yeast isolates[J].J Clin Microbiol,2009,47(9):2912-2917.
[13] van Veen SQ, Claas EC, Kuijper EJ.High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization mass spectrometry in routine medical microbiology laboratory[J].J Clin Microbiol,2010,48(3):900-907.

[1]	王冲, 占萍, 刘维达. MALDI-TOF MS技术在临床病原真菌鉴定中的应用进展[J]. 中国真菌学杂志, 2013, 8(6): 369-373.
[2]	范欣, 王贺, 苏霄翔, 肖盟, 马雪菲, 马超越, 刘旭光, 马然, 吴玥, 陈艳钊, 徐英春. 以分子生物学鉴定结果为“金标准”评估Rapid ID Yeast Plus及API20C AUX对酵母样真菌的鉴定效能[J]. 中国真菌学杂志, 2013, 8(5): 274-280.
[3]	窦红涛, 肖盟, 范欣, 杨启文, 王瑶, 王贺, 刘娟, 谢秀丽, 徐英春. BD Phoenix^TM酵母菌鉴定板对临床分离酵母菌鉴定能力的评估[J]. 中国真菌学杂志, 2013, 8(5): 281-284.
[4]	陈玲, 王红. 侵袭性念珠菌病的抗原抗体检测进展[J]. 中国真菌学杂志, 2013, 8(5): 316-320.
[5]	涂波, 张昕, 李丰衣, 聂为民, 赵敏. 降钙素原在真菌感染诊断中的临床价值[J]. 中国真菌学杂志, 2012, (5): 309-311.
[6]	张超, 廖万清. 基质辅助激光解析电离飞行时间质谱技术在病原真菌检验中的应用进展[J]. 中国真菌学杂志, 2012, (5): 312-315.
[7]	刘春玉, 王红, 任爱民, 杜琨, 胡岚, 李世荣, 文燕, 张淑文. 血浆(1-3)-β-D-葡聚糖诊断深部真菌感染的临床研究[J]. 中国真菌学杂志, 2012, (4): 207-210.
[8]	黄玲莉, 张文宏, 李莉, 章强强. HIV感染患者口咽、鼻腔内真菌分离率的调查[J]. 中国真菌学杂志, 2012, (4): 211-213.
[9]	程娟娟, 潘敏, 韩志君, 葛小丽. 儿童真菌性肠炎患者血清IL-27增高及其意义[J]. 中国真菌学杂志, 2012, 7(1): 24-26.
[10]	高露娟, 余进, 李若瑜. 实时荧光定量PCR在侵袭性曲霉病诊断中的研究进展[J]. 中国真菌学杂志, 2012, 7(1): 59-64.
[11]	杨惠琴, 梅亚宁. 血浆(1,3)-β-D-葡聚糖对深部真菌感染诊断的临床意义[J]. 中国真菌学杂志, 2011, 6(3): 136-140.
[12]	仇萌, 邹先彪. rDNA-ITS序列鉴定深部真菌菌种的研究进展[J]. 中国真菌学杂志, 2011, 6(2): 122-125.
[13]	刘春玉, 王红. 侵袭性真菌感染非培养诊断方法的研究进展[J]. 中国真菌学杂志, 2011, 6(1): 57-60.
[14]	项明洁, 刘锦燕, 倪培华, 张华, 陈华, 倪语星. 重复序列PCR(rep-PCR)指纹分析技术在病原真菌鉴定和分型中的应用[J]. 中国真菌学杂志, 2010, 5(6): 376-380.
[15]	李世荣, 王红, 张淑文. 半乳甘露聚糖对侵袭性曲霉菌感染的诊断价值[J]. 中国真菌学杂志, 2010, 5(6): 381-384.