MALDI-TOF mass signatures for differentiation of yeast species, strain grouping and monitoring of morphogenesis markers

被引:88
作者
Qian, Jiang [1 ,3 ]
Cutler, Jim E. [1 ,2 ]
Cole, Richard B. [3 ]
Cai, Yang [1 ,3 ]
机构
[1] Childrens Hosp, New Orleans Res Inst Children, New Orleans, LA 70118 USA
[2] LSU Hlth Sci Ctr, Dept Pediat & Microbiol, New Orleans, LA 70112 USA
[3] Univ New Orleans, Dept Chem, New Orleans, LA 70148 USA
关键词
mass signature; MALDI-TOF mass spectrometry; alcohol fixation; yeast differentiation; principal component analysis;
D O I
10.1007/s00216-008-2288-1
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) is demonstrated to be a potentially useful tool for the rapid identification of yeasts, the grouping of Candida albicans strains, and the monitoring of germ tube-specific markers. Co-crystallized with sinapinic acid as the MALDI matrix, intact yeast cells yielded a sufficient number of medium-sized ions (4-15 kDa) in MALDI mass spectra to provide "mass signatures" that were diagnostic of strain type. For most isolates, the mass signatures were affected by the growth medium, length of incubation and the cell preparation method. While the overall past success of this methodology for fungal cells has been relatively low compared to its application to bacteria, fixing the yeast cells in 50% methanol inactivated the cells, reduced cell aggregation in aqueous suspension solution, and more importantly, it significantly improved the mass signature quality. This simple but critical advance in sample treatment improved mass spectrometric signal-to-noise ratios and allowed the identification of yeasts by a mass signature approach. Under optimized conditions, Candida species (C. albicans, C. glabrata, C. krusei, C. kefyr), Aspergillus species (A. terreus, A. fumigatus, A. syndowii) and other yeast genera (Cryptococcus neoformans, Saccharomyces cerevisiae and a Rhodotorula sp.) could be distinguished. Within the C. albicans species, several common ions in the m/z 5,000-10,000 range were apparent in the mass spectra of all tested strains. In addition to shared ions, the mass spectra of individual C. albicans strains permitted grouping of the strains. Principal component analysis (PCA) was employed to confirm spectral reproducibility and C. albicans strain grouping by mass signatures. Finally, C. albicans germ tubes produced MALDI-TOF mass signatures that differed from yeast forms of this species. This is a rapid, sensitive and simple method for identifying yeasts, grouping strains and following the morphogenesis of C. albicans.
引用
收藏
页码:439 / 449
页数:11
相关论文
共 63 条
[1]   Characterization of protein biomarkers desorbed by MALDI from whole fungal cells [J].
Amiri-Eliasi, BJ ;
Fenselau, C .
ANALYTICAL CHEMISTRY, 2001, 73 (21) :5228-5231
[2]  
Arnold RJ, 1998, RAPID COMMUN MASS SP, V12, P630, DOI 10.1002/(SICI)1097-0231(19980529)12:10<630::AID-RCM206>3.3.CO
[3]  
2-S
[4]  
Beavis R C, 1989, Rapid Commun Mass Spectrom, V3, P432, DOI 10.1002/rcm.1290031207
[5]   RAPID, SENSITIVE ANALYSIS OF PROTEIN MIXTURES BY MASS-SPECTROMETRY [J].
BEAVIS, RC ;
CHAIT, BT .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (17) :6873-6877
[6]   A forkhead transcription factor is important for true hyphal as well as yeast morphogenesis in Candida albicans [J].
Benson, ES ;
Filler, SC ;
Berman, J .
EUKARYOTIC CELL, 2002, 1 (05) :787-798
[7]   Anionic adducts of oligosaccharides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry [J].
Cai, Y ;
Jiang, YJ ;
Cole, RB .
ANALYTICAL CHEMISTRY, 2003, 75 (07) :1638-1644
[8]   Cell wall and secreted proteins of Candida albicans:: Identification, function, and expression [J].
Chaffin, WL ;
López-Ribot, JL ;
Casanova, M ;
Gozalbo, D ;
Martinez, JP .
MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 1998, 62 (01) :130-+
[9]   Characterization of intact Penicillium spores by matrix-assisted laser desorption/ionization mass spectrometry [J].
Chen, HY ;
Chen, YC .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2005, 19 (23) :3564-3568
[10]   Invasive Saccharomyces infection:: A comprehensive review [J].
Enache-Angoulvant, A ;
Hennequin, C .
CLINICAL INFECTIOUS DISEASES, 2005, 41 (11) :1559-1568