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. 2015 Feb 6;9(2):e0003473.
doi: 10.1371/journal.pntd.0003473. eCollection 2015 Feb.

Detection of Rickettsia spp in ticks by MALDI-TOF MS

Affiliations

Detection of Rickettsia spp in ticks by MALDI-TOF MS

Amina Yssouf et al. PLoS Negl Trop Dis. .

Abstract

Background: Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been shown to be an effective tool for the rapid identification of arthropods, including tick vectors of human diseases.

Methodology/principal findings: The objective of the present study was to evaluate the use of MALDI-TOF MS to identify tick species, and to determine the presence of rickettsia pathogens in the infected Ticks. Rhipicephalus sanguineus and Dermacentor marginatus Ticks infected or not by R. conorii conorii or R. slovaca, respectively, were used as experimental models. The MS profiles generated from protein extracts prepared from tick legs exhibited mass peaks that distinguished the infected and uninfected Ticks, and successfully discriminated the Rickettsia spp. A blind test was performed using Ticks that were laboratory-reared, collected in the field or removed from patients and infected or not by Rickettsia spp. A query against our in-lab arthropod MS reference database revealed that the species and infection status of all Ticks were correctly identified at the species and infection status levels.

Conclusions/significance: Taken together, the present work demonstrates the utility of MALDI-TOF MS for a dual identification of tick species and intracellular bacteria. Therefore, MALDI-TOF MS is a relevant tool for the accurate detection of Rickettsia spp in Ticks for both field monitoring and entomological diagnosis. The present work offers new perspectives for the monitoring of other vector borne diseases that present public health concerns.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Comparison of MALDI-TOF MS profiles of Ticks infected or not by Rickettsia spp.
Representative spectra from biological replicates of Rh. sanguineus (A, B), Rh. sanguineus infected by R. conorii conorii (C, D), D. marginatus (E, F) and D. marginatus infected by R. slovaca (G, H) were aligned using Flex analysis 3.3 software. a.u., arbitrary units; m/z, mass-to-charge ratio.
Fig 2
Fig 2. Alignment of MALDI-TOF MS profiles of Rickettsia strains and tick species infected or not by Rickettsia using Flex analysis 3.3 software.
Representative spectra of a purified R. conorii conorii strain (A), Rh. sanguineus (B), Rh. sanguineus infected by R. conorii conorii (C), a purified R. slovaca strain (D), D. marginatus (E) and D. marginatus infected by R. slovaca (F) are presented. a.u., arbitrary units; m/z, mass-to-charge ratio.
Fig 3
Fig 3. Location of discriminating peak masses shared between the spectra acquired from Rickettsia-infected specimens and the corresponding Rickettsia strain using Flex analysis software 3.3.
The alignment spectra comparing the infected specimen and the corresponding strain spectra are shown in detail. (A) R. conorii conorii shared discriminating peak masses located on the MS profile of Rh. sanguineus infected by R. conorii conorii. (B) R. slovaca shared discriminating peak masses located on the MS profile of D. marginatus infected by R. slovaca.

References

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