Chronic Chagas Disease Diagnosis: A Comparative Performance of Commercial Enzyme Immunoassay Tests

Santos, Fred Luciano Neves; Souza, Wayner Vieira de; Barros, Michelle da Silva; Nakazawa, Mineo; Krieger, Marco Aurélio; Gomes, Yara de Miranda

doi:10.4269/ajtmh.15-0820

Cited by 38 publications

(39 citation statements)

References 28 publications

(29 reference statements)

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“…Samples from individuals with dengue virus (n ϭ 50), hepatitis B virus (n ϭ 160), hepatitis C virus (n ϭ 98), human immunodeficiency virus (n ϭ 144), human T-cell lymphotropic virus (n ϭ 109), leishmaniasis (n ϭ 18), leptospirosis (n ϭ 92), rubella virus (n ϭ 15), measles (n ϭ 21), and syphilis (n ϭ 144) were used to assess cross-reactivity between the IBMP chimeras and proteins associated with unrelated diseases. Before LMA analysis, all sera were reevaluated using 2 commercial ELISAs, namely, the Imuno-ELISA Chagas test (Wama Diagnostica, São Paulo, Brazil; batch 14D061) and the ELISA Chagas III test (BIOSChile, Ingeniaría Genética S.A., Santiago, Chile; batch 1F130525) (27). Each sample was assigned a numeric code in the laboratory to ensure a blinded analysis.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2017

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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 antigens showed excellent diagnostic performance in enzyme-linked immunosorbent assays. Considering that next-generation platforms offer improved CD diagnostic accuracy with different-specific recombinant antigens, we assessed the performance of these chimeras in liquid microarrays (LMAs). The chimeric proteins were expressed in 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- 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. , a pathogen with high similarity to, 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.

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Section: Methodsmentioning

confidence: 99%

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

et al. 2017

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“…Sample selection was based on positivity or negativity by two serological tests for CD: Imuno-ELISA Chagas (Wama Diagnóstica, São Palo, Brazil, batch 14D061), which is based on recombinant antigens; and ELISA Chagas III (BIOSChile, Ingeniería Genética S.A., Santiago, Chile, batch 1F130525), which uses whole extracts of T . cruzi strains Mn and Tulahuen as antigens [ 33 ]. Samples with disagreeing results between both tests or judged to be inconclusive in one of them were excluded.…”

Section: Methodsmentioning

confidence: 99%

Performance Assessment of Four Chimeric Trypanosoma cruzi Antigens Based on Antigen-Antibody Detection for Diagnosis of Chronic Chagas Disease

Santos¹,

Celedón²,

Zanchin³

et al. 2016

PLoS ONE

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The performance of serologic tests in chronic Chagas disease diagnosis largely depends on the type and quality of the antigen preparations that are used for detection of anti-Trypanosoma cruzi antibodies. Whole-cell T. cruzi extracts or recombinant proteins have shown variation in the performance and cross-reactivity. Synthetic chimeric proteins comprising fragments of repetitive amino acids of several different proteins have been shown to improve assay performances to detect Chagasic infections. Here, we describe the production of four chimeric T. cruzi proteins and the assessment of their performance for diagnostic purposes. Circular Dichroism spectra indicated the absence of well-defined secondary structures, while polydispersity evaluated by Dynamic Light Scattering revealed only minor aggregates in 50 mM carbonate-bicarbonate (pH 9.6), demonstrating that it is an appropriate buffering system for sensitizing microplates. Serum samples from T. cruzi-infected and non-infected individuals were used to assess the performance of these antigens for detecting antibodies against T. cruzi, using both enzyme-linked immunosorbent assay and a liquid bead array platform. Performance parameters (AUC, sensitivity, specificity, accuracy and J index) showed high diagnostic accuracy for all chimeric proteins for detection of specific anti-T. cruzi antibodies and differentiated seropositive individuals from those who were seronegative. Our data suggest that these four chimeric proteins are eligible for phase II studies.

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“…However, the performance of these assays varies greatly because of the antigen preparations used to detect the anti– T. cruzi antibodies, in addition to genetic heterogeneity among circulating strains. 16 , 18 Because no reference standard is currently available for CD, the World Health Organization has advocated the use of two tests in parallel for serodiagnostic verification. Under these circumstances, the diagnostic accuracy of novel methodologies cannot be evaluated in an unbiased manner, because the comparison of results implies the use of an imperfect reference standard.…”

Section: Introductionmentioning

confidence: 99%

Highly Accurate Chimeric Proteins for the Serological Diagnosis of Chronic Chagas Disease: A Latent Class Analysis

Santos

Campos

Amorim

et al. 2018

The American Journal of Tropical Medicine and Hygiene

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Abstract.The existence of an imperfect reference standard presents complications when evaluating the unbiased performance of novel diagnostic techniques. This is especially true in the absence of a gold standard, as is the case in chronic Chagas disease (CD) diagnosis. To circumvent this constraint, we elected to use latent class analysis (LCA). Previously, our group demonstrated the high performance of four Trypanosoma cruzi–chimeric proteins (Molecular Biology Institute of Paraná [IBMP]-8.1, -8.2, -8.3, and -8.4) for CD diagnosis using several distinct immunoassays. Although commercial tests had previously been established as a reference standard, the diagnostic performance of these chimeric antigens could present bias because these tests fail to produce 100% accurate results. Thus, we used LCA to assess the performance of these IBMP chimeric antigens in chronic CD diagnosis. Using the LCA model as a gold standard, sensitivity and specificity values ranged from 93.5% to 99.4% and 99.6% to 100%, respectively. The accuracy values were 96.2% for IBMP-8.2, approximately 98% for IBMP-8.1 and IBMP-8.3, and nearly 100% for IBMP-8.4. For IBMP-8.1 and IBMP-8.2, higher positive predictive values were associated with increases in hypothetical prevalence. Similarly, higher hypothetical prevalence resulted in lower negative predictive values for IBMP-8.1, IBMP-8.2, and IBMP-8.3. In addition, samples with serodiscordant results from commercial serological tests were analyzed using LCA. Molecular Biology Institute of Paraná -8.1 demonstrated potential for use in confirmatory testing with regard to samples with inconsistent results. Moreover, our findings further confirmed the remarkable performance of the IBMP-8.4 antigen to diagnose chronic CD in both endemic and non-endemic areas.

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Chronic Chagas Disease Diagnosis: A Comparative Performance of Commercial Enzyme Immunoassay Tests

Cited by 38 publications

References 28 publications

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

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

Performance Assessment of Four Chimeric Trypanosoma cruzi Antigens Based on Antigen-Antibody Detection for Diagnosis of Chronic Chagas Disease

Highly Accurate Chimeric Proteins for the Serological Diagnosis of Chronic Chagas Disease: A Latent Class Analysis

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