Format

Send to

Choose Destination
J Clin Microbiol. 2017 Oct;55(10):2934-2945. doi: 10.1128/JCM.00851-17. Epub 2017 Jul 19.

Performance Assessment of a Trypanosoma cruzi Chimeric Antigen in Multiplex Liquid Microarray Assays.

Author information

1
Gonçalo Moniz Institute, FIOCRUZ-BA, Salvador, Bahia, Brazil fred.santos@bahia.fiocruz.br.
2
Programa Integrado de Doença de Chagas, Oswaldo Cruz Institute, FIOCRUZ-RJ, Rio de Janeiro, Rio de Janeiro, Brazil.
3
Molecular Biology Institute of Paraná, Curitiba, Paraná, Brazil.
4
Carlos Chagas Institute, FIOCRUZ-PR, Curitiba, Paraná, Brazil.
5
Aggeu Magalhães Institute, FIOCRUZ-PE, Recife, Pernambuco, Brazil.

Abstract

Diagnosing chronic Chagas disease (CD) requires antibody-antigen detection methods, which are traditionally based on enzymatic assay techniques whose performance depend on the type and quality of antigen used. Previously, 4 recombinant chimeric proteins from the Instituto de Biologia Molecular do Paraná (IBMP-8.1 to 8.4) comprising immuno-dominant regions of diverse Trypanosoma cruzi antigens showed excellent diagnostic performance in enzyme-linked immunosorbent assays. Considering that next-generation platforms offer improved CD diagnostic accuracy with different T. cruzi-specific recombinant antigens, we assessed the performance of these chimeras in liquid microarrays (LMAs). The chimeric proteins were expressed in Escherichia coli and purified by chromatography. Sera from 653 chagasic and 680 healthy individuals were used to assess the performance of these chimeras in detecting specific anti-T. cruzi antibodies. Accuracies ranged from 98.1 to 99.3%, and diagnostic odds ratio values were 3,548 for IBMP-8.3, 4,826 for IBMP-8.1, 7,882 for IBMP-8.2, and 25,000 for IBMP-8.4. A separate sera bank (851 samples) was employed to assess cross-reactivity with other tropical diseases. Leishmania, a pathogen with high similarity to T. cruzi, showed cross-reactivity rates ranging from 0 to 2.17%. Inconclusive results were negligible (0 to 0.71%). Bland-Altman and Deming regression analysis based on 200 randomly selected CD-positive and negative samples demonstrated interchangeability with respect to CD diagnostic performance in both singleplex and multiplex assays. Our results suggested that these chimeras can potentially replace antigens currently used in commercially available assay kits. Moreover, the use of multiplex platforms, such as LMA assays employing 2 or more IBMP antigens, would abrogate the need for 2 different testing techniques when diagnosing CD.

KEYWORDS:

Trypanosoma cruzi; chimeric antigens; human Chagas disease; liquid microarray; singleplex and multiplex assays

PMID:
28724556
PMCID:
PMC5625379
DOI:
10.1128/JCM.00851-17
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center