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. 2018 Jan 4;11(1):11.
doi: 10.1186/s13071-017-2597-3.

MALDI-TOF MS as a new tool for the identification of Dientamoeba fragilis

Adriana Calderaro  1 , Mirko Buttrini  2 , Sara Montecchini  2 , Sabina Rossi  2 , Giovanna Piccolo  2 , Maria Cristina Arcangeletti  2 , Maria Cristina Medici  2 , Carlo Chezzi  2 , Flora De Conto  2
Affiliations

MALDI-TOF MS as a new tool for the identification of Dientamoeba fragilis

Adriana Calderaro et al. Parasit Vectors. .

Abstract

Background: In this study for the first time, a Dientamoeba fragilis protein profile by MALDI-TOF MS was created in order to identify specific markers for the application of this technology in the laboratory diagnosis of dientamoebiasis. In particular, one D. fragilis reference strain was used to create a reference spectrum and 14 clinical isolates to verify the reliability of the obtained results.

Results: While 15 peaks were found to be discriminating between the reference strain and the culture medium used, six peaks, observed in all the 14 strains tested, were considered as markers able to identify D. fragilis.

Conclusions: In our hands, MALDI-TOF MS technology was demonstrated as a useful tool to be used in association with or in replacement of the real-time PCR assay for the identification of D. fragilis used in our laboratory on xenic cultures, due to its accuracy, rapidity and low cost.

Keywords: Dientamoeba fragilis; Identification; MALDI-TOF mass spectrometry.

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Conflict of interest statement

Ethics approval and consent to participate

The samples analyzed in this and previous studies were obtained by the University Hospital of Parma for routine diagnosis purposes. No approval by the Institutional Review Board was required because the laboratory diagnosis results had been reported in the medical records of the patients as a diagnostic answer to a clinical suspicion of intestinal parasitosis. Ethical approval at the University Hospital of Parma is required only in cases where the clinical samples are to be used for applications other than diagnosis.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Comparison of the spectra obtained for the D. fragilis No. 3313 reference strain (DF3313) and the Robinson’s medium alone (a) and cluster analysis by principal components analysis (PCA) (b) of the replicates of the DF3313 (red) and of the Robinson’s medium alone (green)
Fig. 2
Fig. 2
Examples of peaks (5433, 5041 and 5087 Da) discriminating the D. fragilis No. 3313 reference strain (green) from the Robinson’s medium alone (red) on the basis of the average spectra obtained
Fig. 3
Fig. 3
Analysis of the spectra obtained for the D. fragilis No. 3313 reference strain at different trophozoite concentrations (106 trophozoites/ml in green, 105 trophozoites/ml in blue, 104 trophozoites/ml in yellow, 103 trophozoites/ml in purple) in comparison to the average reference spectrum in red. a Examples of the presence/absence of two discriminating peaks (5041 and 5087 Da). b Cluster analysis by “2D distribution view” of the first two best separating peaks (left) (the ellipses represent the standard deviation of each concentration average of the peak area/intensity) and by principal components analysis (PCA) (right)
Fig. 4
Fig. 4
Analysis of the spectra obtained from the two D. fragilis experimentally seeded fecal samples. a Example of discriminating peaks found in the fecal sample experimentally seeded with the D. fragilis No. 3313 reference strain at the 106 trophozoites/ml concentration (green) in comparison to the average reference spectrum (red). b Cluster analysis by principal components analysis (PCA) of the fecal sample experimentally seeded with the D. fragilis No. 1686 clinical isolate at the 106 trophozoites/ml concentration (blue) in comparison to the D. fragilis No.1686 strain at the same concentration without feces (green) and to the average reference spectrum (red)
See this image and copyright information in PMC

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