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Analysis of Four Human Microsporidian Isolates by MALDI-TOF Mass Spectrometry

Authors

  • HERCULES MOURA,

    Corresponding author
    1. Atlanta Research and Education Foundation, Rio de Janeiro
    2. Universidade do Estado do Rio de Janeiro & HEC-F1OCRUZ, Rio de Janeiro, Brazil
    3. Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30341
      Corresponding Author: H. Moura—Telephone number: 770-488-7843; FAX number: 770-488-4609; E-mail: Hmoura@cdc.gov
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  • MARIA OSPINA,

    1. Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA
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  • ADRIAN R. WOOLFITT,

    1. Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA
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  • JOHN R. BARR,

    1. Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, Georgia 30341, USA
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  • GOVINDA S. VISVESVARA

    1. Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30341
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Corresponding Author: H. Moura—Telephone number: 770-488-7843; FAX number: 770-488-4609; E-mail: Hmoura@cdc.gov

Abstract

ABSTRACT. Spores of four species of microsporidia isolated from humans were analyzed by matrix-assisted laser desorption ioni-zation time of flight mass spectrometry (MALDI-TOF MS) and specific biomarkers were found for each. The microsporidia analyzed included three species, Encephalitozoon cuniculi, Encephalitozoon hellem, and Encephalitozoon intestinalis and the fourth organism is the recently described Brachiola algerae. Whole spores, spore shells, and soluble fractions were applied directly to the MALDI target without further purification steps. MALDI-TOF MS analysis of both whole spores and soluble fractions of the four isolates revealed a group of unique, characteristic, and reproducible spectral markers in the mass range of 2,000-8,000 Da. Statistical analysis of the averaged centroided masses uncovered two distinct sets of unique peptides or biomarkers, one originated from whole spores and the other from soluble fractions, that can differentiate the four microsporidian species studied. MALDI-TOF MS analysis of whole organisms is a rapid, sensitive, and specific option to characterize microsporidian isolates and has the potential for several applications in parasi-tology.

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