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a computational pipeline for diagnostic biomarker discovery in the human pathogen trypanosoma cruzi

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cruzi genome sequence to identify new peptidic diag-nostic biomarkers.. Methods An integrative bioinformatic strategy was adopted to prioritize peptidic antigens with low cross-reactivit

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O R A L P R E S E N T A T I O N Open Access

A computational pipeline for diagnostic

biomarker discovery in the human pathogen

Trypanosoma cruzi

Santiago J Carmona1*, Paula Sartor2, Maria Susana Leguizamón2, Oscar Campetella1, Fernán Agüero1

From Sixth International Society for Computational Biology (ISCB) Student Council Symposium

Boston, MA, USA 9 July 2010

Background

The protozoan parasiteTrypanosoma cruzi is the

causa-tive agent of Chagas’ disease, endemic in 18 countries in

Central and South America Transmission also occurs in

non-endemic countries by way of blood transfusion and

organ transplantation Diagnosis of American

trypanoso-miasis is based on the detection of antibodies directed

against T cruzi antigens In this work we mined the

T cruzi genome sequence to identify new peptidic diag-nostic biomarkers

Methods

An integrative bioinformatic strategy was adopted to prioritize peptidic antigens with low cross-reactivity in the genome of T cruzi A computational pipeline was developed to assess a set of molecular properties on

* Correspondence: sjcarmona@gmail.com

Full list of author information is available at the end of the article

Figure 1 Example sequence profile generated by the pipeline using BioPerl Vertical boxes in the plot represent overlapping 12-residue peptides, and its height and colour, the resulting score based on the mapped features shown below.

Carmona et al BMC Bioinformatics 2010, 11(Suppl 10):O11

http://www.biomedcentral.com/1471-2105/11/S10/O11

© 2010 Carmona et al; licensee BioMed Central Ltd This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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each protein from the referenceT cruzi genome, such

as subcellular localization or expression level (by mass

spec evidence, number of gene copies and synonymous

codon usage bias) At a higher resolution, a set of local

properties were evaluated, such as repetitive motifs,

dis-order (structured vs natively unstructured regions),

trans-membrane spans, glycosylation sites,

polymorph-isms (conserved vs divergent regions), predicted B-cell

epitopes, sequence similarity against human proteins

andLeishmania (potential cross-reacting species) (Figure

1) A scoring function based on these properties was

used to rank each of the ~10 million 12-residue

overlap-ping peptides in which the ~ 22,000 T cruzi proteins

can be virtually fragmented Experimental validation of

predicted epitopes was performed with peptide

microar-rays, screened using pooled sera from human chagasic

patients and controls

Results

We show that our integrative method outperforms

alter-native antigen prioritizations based on individual

proper-ties (such as B-cell epitope predictors alone) Our

genome-wide prioritization uncovered more than 300

pro-mising biomarker candidates 200 high-scoring peptides

corresponding mostly to hypothetical proteins were

selected for immunological validation, along with 40

pep-tides derived from previously validated B-cell epitopes and

an additional set of 40 low-scoring peptides as controls

Preliminary results based on microarray images revealed

that ~25% (49/200) of the candidate peptides reacted

spe-cifically against the positive sera pools assayed

Conclusion

The developed bioinformatic approach proved to be

successful, leading from a genome-wide prioritization to

the identification of novel peptidic antigens with

diag-nostic potential Moreover, the algorithm may be used

to prioritize biomarkers in other pathogen species

Acknowledgements

This work was funded by Universidad de San Martín (grant PROG07F/1) and

the “Special Programme for Research and Training in Tropical Diseases

(UNICEF/UNDP/World Bank/WHO) ”.

Author details

1 Instituto de Investigaciones Biotecnológicas, Universidad de San Martín, San

Martín, Argentina.2Departamento de Microbiología, Facultad de Medicina,

Universidad de Buenos Aires, Buenos Aires, Argentina.

Published: 7 December 2010

doi:10.1186/1471-2105-11-S10-O11

Cite this article as: Carmona et al.: A computational pipeline for

diagnostic biomarker discovery in the human pathogen Trypanosoma

cruzi BMC Bioinformatics 2010 11(Suppl 10):O11.

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Carmona et al BMC Bioinformatics 2010, 11(Suppl 10):O11

http://www.biomedcentral.com/1471-2105/11/S10/O11

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