Direct identification of trypanosomatids by matrix-assisted laser desorption ionization-time of flight mass spectrometry (DIT MALDI-TOF MS)

Avila, Carla Cristi; Filho, Flávio Guilherme Vaz de Almeida; Palmisano, Giuseppe

doi:10.1002/jms.3763

Cited by 14 publications

(6 citation statements)

References 54 publications

(61 reference statements)

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“…This technique is based on laser ionization of sample proteins after co-crystallization with MALDI-matrix and comparison of the obtained mass spectra with a database of reference spectra [5]. MALDI-TOF MS is currently under development for the study of protozoa with potential use for the identification of Leishmania [22], Plasmodium [23] and trypanosomatids [1]. Few applications, however, have been proposed in the field of helminthology.…”

Section: Introductionmentioning

confidence: 99%

MALDI-TOF mass spectrometry: a new tool for rapid identification of cercariae (Trematoda, Digenea)

Huguenin¹,

Depaquit²,

Villena³

et al. 2019

Parasite

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Identification of cercariae was long based on morphological and morphometric features, but these approaches remain difficult to implement and require skills that have now become rare. Molecular tools have become the reference even though they remain relatively time-consuming and expensive. We propose a new approach for the identification of cercariae using MALDI-TOF mass spectrometry. Snails of different genera (Radix, Lymnaea, Stagnicola, Planorbis, and Anisus) were collected in the field to perform emitting tests in the laboratory. The cercariae they emitted (Trichobilharzia anseri, Diplostomum pseudospathaceum, Alaria alata, Echinostoma revolutum, Petasiger phalacrocoracis, Tylodelphys sp., Australapatemon sp., Cotylurus sp., Posthodiplostomum sp., Parastrigea sp., Echinoparyphium sp. and Plagiorchis sp.) were characterized by sequencing the D2, ITS2 and ITS1 domains of rDNA, and by amplification using specific Alaria alata primers. A sample of each specimen, either fresh or stored in ethanol, was subjected to a simple preparation protocol for MALDI-TOF analysis. The main spectral profiles were analyzed by Hierarchical Clustering Analysis. Likewise, the haplotypes were analyzed using the maximum likelihood method. Analytical performance and the log-score value (LSV) cut-off for species identification were then assessed by blind testing. The clusters obtained by both techniques were congruent, allowing identification at a species level. MALDI-TOF enables identification at an LSV cut-off of 1.7 without false-positives; however, it requires more data on closely related species. The development of a “high throughput” identification system for all types of cercariae would be of considerable interest in epidemiological surveys of trematode infections.

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Section: Introductionmentioning

confidence: 99%

MALDI-TOF mass spectrometry: a new tool for rapid identification of cercariae (Trematoda, Digenea)

Huguenin¹,

Depaquit²,

Villena³

et al. 2019

Parasite

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“…Due to that, novel methods identify and characterize DTUs can offer opportunities to understand the DTUs diversity, link with their phenotype and provide another tool for molecular epidemiology. Recently, MALDI-based strategy was developed for the direct identification of trypanosomatids based on the MS profile, named DIT-MALDI TOF [ 39 ]. However, this method does not have the resolution to discriminate DTUs using the MS profile.…”

Section: Discussionmentioning

confidence: 99%

Development of a Trypanosoma cruzi strain typing assay using MS2 peptide spectral libraries (Tc-STAMS2)

et al. 2018

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BackgroundChagas disease also known as American trypanosomiasis is caused by the protozoan Trypanosoma cruzi. Over the last 30 years, Chagas disease has expanded from a neglected parasitic infection of the rural population to an urbanized chronic disease, becoming a potentially emergent global health problem. T. cruzi strains were assigned to seven genetic groups (TcI-TcVI and TcBat), named discrete typing units (DTUs), which represent a set of isolates that differ in virulence, pathogenicity and immunological features. Indeed, diverse clinical manifestations (from asymptomatic to highly severe disease) have been attempted to be related to T.cruzi genetic variability. Due to that, several DTU typing methods have been introduced. Each method has its own advantages and drawbacks such as high complexity and analysis time and all of them are based on genetic signatures. Recently, a novel method discriminated bacterial strains using a peptide identification-free, genome sequence-independent shotgun proteomics workflow. Here, we aimed to develop a Trypanosoma cruzi Strain Typing Assay using MS/MS peptide spectral libraries, named Tc-STAMS2.Methods/Principal findingsThe Tc-STAMS2 method uses shotgun proteomics combined with spectral library search to assign and discriminate T. cruzi strains independently on the genome knowledge. The method is based on the construction of a library of MS/MS peptide spectra built using genotyped T. cruzi reference strains. For identification, the MS/MS peptide spectra of unknown T. cruzi cells are identified using the spectral matching algorithm SpectraST. The Tc-STAMS2 method allowed correct identification of all DTUs with high confidence. The method was robust towards different sample preparations, length of chromatographic gradients and fragmentation techniques. Moreover, a pilot inter-laboratory study showed the applicability to different MS platforms.Conclusions and significanceThis is the first study that develops a MS-based platform for T. cruzi strain typing. Indeed, the Tc-STAMS2 method allows T. cruzi strain typing using MS/MS spectra as discriminatory features and allows the differentiation of TcI-TcVI DTUs. Similar to genomic-based strategies, the Tc-STAMS2 method allows identification of strains within DTUs. Its robustness towards different experimental and biological variables makes it a valuable complementary strategy to the current T. cruzi genotyping assays. Moreover, this method can be used to identify DTU-specific features correlated with the strain phenotype.

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“…Bruker BioTyper based database compilation were also reported for the food nematode Trichinella (Mayer-Scholl et al, 2016), Trichomonas vaginalis (Calderaro et al, 2016), Acanthamoeba spp. (Del Chierico et al, 2016) using a database for direct identification of trypanosomatids (Avila et al, 2016). …”

Section: Advances In Invertebrate Pests Parasite and Vector Profilingmentioning

confidence: 99%

MALDI-TOF MS Profiling-Advances in Species Identification of Pests, Parasites, and Vectors

Murugaiyan

Roesler

2017

Front. Cell. Infect. Microbiol.

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Invertebrate pests and parasites of humans, animals, and plants continue to cause serious diseases and remain as a high treat to agricultural productivity and storage. The rapid and accurate species identification of the pests and parasites are needed for understanding epidemiology, monitoring outbreaks, and designing control measures. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has emerged as a rapid, cost effective, and high throughput technique of microbial species identification in modern diagnostic laboratories. The development of soft ionization techniques and the release of commercial pattern matching software platforms has resulted in the exponential growth of applications in higher organisms including parasitology. The present review discusses the proof-of-principle experiments and various methods of MALDI MS profiling in rapid species identification of both laboratory and field isolates of pests, parasites and vectors.

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Direct identification of trypanosomatids by matrix-assisted laser desorption ionization-time of flight mass spectrometry (DIT MALDI-TOF MS)

Cited by 14 publications

References 54 publications

MALDI-TOF mass spectrometry: a new tool for rapid identification of cercariae (Trematoda, Digenea)

MALDI-TOF mass spectrometry: a new tool for rapid identification of cercariae (Trematoda, Digenea)

Development of a Trypanosoma cruzi strain typing assay using MS2 peptide spectral libraries (Tc-STAMS2)

MALDI-TOF MS Profiling-Advances in Species Identification of Pests, Parasites, and Vectors

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