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Refining wet lab experiments with in silico searches: a rational quest for diagnostic peptides in visceral leishmaniasis

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dc.contributor.author Bremer Hinckel, B. C. en_US
dc.contributor.author Marlais, T. en_US
dc.contributor.author Airs, S. en_US
dc.contributor.author Bhattacharyya, T. en_US
dc.contributor.author Imamura, H. en_US
dc.contributor.author Dujardin, J. C. en_US
dc.contributor.author El-Safi, S. en_US
dc.contributor.author Singh, O. P. en_US
dc.contributor.author Sundar, S. en_US
dc.contributor.author Falconar, A. K. en_US
dc.contributor.author Andersson, B. en_US
dc.contributor.author Litvinov, S. en_US
dc.contributor.author Miles, M. A. en_US
dc.contributor.author Mertens, P. en_US
dc.date.accessioned 2019-06-04T12:20:34Z
dc.date.available 2019-06-04T12:20:34Z
dc.date.issued 2019 en_US
dc.identifier.issn 1935-2727 en_US
dc.identifier.doi http://dx.doi.org/10.1017/10.1371/journal.pntd.0007353 en_US
dc.identifier.other ITG-B4A; ITG-B5A; DBM; U-MOLPAR; JIF; DOI; CPDF; Abstract; ITMPUB; DSPACE67 en_US
dc.identifier.uri http://hdl.handle.net/10390/10616
dc.description.abstract BACKGROUND: The search for diagnostic biomarkers has been profiting from a growing number of high quality sequenced genomes and freely available bioinformatic tools. These can be combined with wet lab experiments for a rational search. Improved, point-of-care diagnostic tests for visceral leishmaniasis (VL), early case detection and surveillance are required. Previous investigations demonstrated the potential of IgG1 as a biomarker for monitoring clinical status in rapid diagnostic tests (RDTs), although using a CLA as capturing antigen. Replacing the CLA by specific antigens would lead to more robust RDTs. METHODOLOGY: Immunoblots revealed L. donovani protein bands detected by IgG1 from VL patients. Upon confident identification of these antigens by mass spectrometry (MS), we searched for evidence of constitutive protein expression and presence of antigenic domains or high accessibility to B-cells. Selected candidates had their linear epitopes mapped with in silico algorithms. Multiple high-scoring predicted epitopes from the shortlisted proteins were screened in peptide arrays. The most promising candidate was tested in RDT prototypes using VL and nonendemic healthy control (NEHC) patient sera. RESULTS: Over 90% of the proteins identified from the immunoblots did not satisfy the selection criteria and were excluded from the downstream epitope mapping. Screening of predicted epitope peptides from the shortlisted proteins identified the most reactive, for which the sensitivity for IgG1 was 84% (95% CI 60-97%) with Sudanese VL sera on RDT prototypes. None of the sera from NEHCs were positive. CONCLUSION: We employed in silico searches to reduce drastically the output of wet lab experiments, focusing on promising candidates containing selected protein features. By predicting epitopes in silico we screened a large number of peptides using arrays, identifying the most promising one, for which IgG1 sensitivity and specificity, with limited sample size, supported this proof of concept strategy for diagnostics discovery, which can be applied to the development of more robust IgG1 RDTs for monitoring clinical status in VL. en_US
dc.language English en_US
dc.relation.uri http://www.ncbi.nlm.nih.gov/pubmed/31059497 en_US
dc.subject Leishmaniasis en_US
dc.subject Visceral en_US
dc.subject Protozoal diseases en_US
dc.subject Diagnostic tests en_US
dc.subject Peptides en_US
dc.subject Experimental en_US
dc.title Refining wet lab experiments with in silico searches: a rational quest for diagnostic peptides in visceral leishmaniasis en_US
dc.type Article-E en_US
dc.citation.issue 5 en_US
dc.citation.jtitle PLoS Neglected Tropical Diseases en_US
dc.citation.volume 13 en_US
dc.citation.pages e0007353 en_US
dc.citation.abbreviation PLoS Negl Trop Dis en_US


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