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Author Spotlight: Studying Behavior of Acanthamoeba to Develop Targeted Strategies for Preventing Acanthamoeba Keratitis
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Protein profiling of Acanthamoeba species using MALDI-TOF MS for specific identification of Acanthamoeba genotype

Kirti Megha1, Megha Sharma1, Amit Gupta2

  • 1Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.

Parasitology Research
|January 19, 2018
PubMed

Insights

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can accurately identify Acanthamoeba genotypes. This method differentiates clinical isolates from environmental contaminants, aiding infection control.

Area of Science:

  • Microbiology
  • Medical Diagnostics

Background:

  • Acanthamoeba species are widespread environmental protozoa capable of causing serious human infections.
  • Genotypic variation within Acanthamoeba influences disease severity and treatment outcomes.
  • Accurate genotyping is crucial for effective infection control strategies.

Purpose of the Study:

  • To evaluate the utility of MALDI-TOF MS for Acanthamoeba genotyping.
  • To develop a MALDI-TOF MS database for differentiating Acanthamoeba genotypes.
  • To compare MALDI-TOF MS identification with 18S rDNA sequencing.

Main Methods:

  • Twenty-five Acanthamoeba isolates were characterized using 18S rDNA sequencing.
  • A reference database was created using MALDI-TOF MS spectra from known genotypes.
  • Coded Acanthamoeba samples were analyzed using MALDI-TOF MS and compared to the database.

Main Results:

  • Five Acanthamoeba genotypes (T3, T4, T5, T10, T11) were identified.
  • MALDI-TOF MS analysis revealed distinct genus-specific and genotype-specific spectral peaks.
  • Spectral patterns were reproducible, with clinical and environmental isolates of the same genotype showing variations.
  • MALDI-TOF MS achieved 92% concordance with 18S rDNA sequencing.

Conclusions:

  • Optimized MALDI-TOF MS is a reliable method for Acanthamoeba spp. genotyping.
  • This technique can differentiate clinical isolates from environmental contaminants.
  • MALDI-TOF MS offers a rapid and accurate tool for Acanthamoeba identification in clinical settings.
Keywords:
18S rDNAAmoebic keratitisGenotypingGranulomatous amoebic encephalitis

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