Use of an enzyme-linked immunosorbent assay that utilizes the Tc13Tul antigen of Trypanosoma cruzi to monitor patients after treatment with benznidazole

Santamaría, Ana; Rissio, Ana María De; Riarte, Adelina; Garavaglia, Patricia Andrea; Bruballa, Andrea Cecilia; Rodríguez, Marisa; Irazú, Lucía; Ruiz, Andrés M.; García, Gabriela Andrea

doi:10.1016/j.diagmicrobio.2013.02.028

Cited by 6 publications

(7 citation statements)

References 41 publications

(66 reference statements)

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“…These data are in accordance with results from a phase II study, where these same proteins were tested by ELISA (16), thereby indicating the high discriminative power these antigens potentially possess with respect to other diagnostic platforms. Most importantly, these IBMP chimeric proteins provided much better AUC values than did T. cruzi cell lysates, single recombinant proteins, or other recombinant chimeric proteins commonly used in diagnostic kits (17,18). In addition, differences greater than 4.40 were seen between the RI signals from the positive and negative samples for all proteins, providing further evidence of their high discriminatory capability.…”

Section: Discussionmentioning

confidence: 85%

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: Discussionmentioning

confidence: 85%

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

et al. 2017

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“…T . cruzi- infected patients demonstrate a large diversity in clinical forms as well as in the pattern of antibody responses, i.e., it is uncommon to find a single antigen that reacts with all seropositive sample [ 26 ] [ 29 ] [ 30 ]. Indeed, we previously described that a minimum of 4 antigens were required to confirm a sample as positive [ 26 ].…”

Section: Discussionmentioning

confidence: 99%

A novel antibody surrogate biomarker to monitor parasite persistence in Trypanosoma cruzi-infected patients

Zrein¹,

Granjon²,

Gueyffier³

et al. 2018

PLoS Negl Trop Dis

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BackgroundTrypanosoma cruzi parasite, the causative agent of Chagas disease, infects about six million individuals in more than 20 countries. Monitoring parasite persistence in infected individuals is of utmost importance to develop and evaluate treatments to control the disease. Routine screening for infected human individuals is achieved by serological assays; PCR testing to monitor spontaneous or therapy-induced parasitological cure has limitations due to the low and fluctuating parasitic load in circulating blood. The aim of the present study is to evaluate a newly developed antibody profiling assay as an indirect method to assess parasite persistence based on waning of antibodies following spontaneous or therapy-induced clearance of the infection.Methodology/Principal findingsWe designed a multiplex serology assay, an array of fifteen optimized T. cruzi antigens, to evaluate antibody diversity in 1654 serum samples from chronic Chagas patients. One specific antibody response (antibody 3, Ab3) showed a strong correlation with T. cruzi parasite persistence as determined by T. cruzi PCR positive results. High and sustained Ab3 signal was strongly associated with PCR positivity in untreated patients, whereas significant decline in Ab3 signals was observed in BZN-treated patients who cleared parasitemia based on blood PCR results.Conclusion/SignificanceAb3 is a new surrogate biomarker that strongly correlates with parasite persistence in chronic and benznidazole-treated Chagas patients. We hypothesize that Ab3 is induced and maintained by incessant stimulation of the immune system by tissue-based and shed parasites that are not consistently detectable by blood based PCR techniques. Hence, a simple immunoassay measurement of Ab3 could be beneficial for monitoring the infectious status of seropositive patients.

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“…Considering the AUC values reported in the literature, the IBMP proteins provided a higher discrimination power compared to other recombinant chimeric proteins, single recombinant proteins, or cell lysates from T . cruzi strains [38,39]. Furthermore, assays with IBMP proteins yielded a low number of inconclusive results.…”

Section: Discussionmentioning

confidence: 99%

Accuracy of chimeric proteins in the serological diagnosis of chronic chagas disease – a Phase II study

et al. 2017

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BackgroundThe performance of current serologic tests for diagnosing chronic Chagas disease (CD) is highly variable. The search for new diagnostic markers has been a constant challenge for improving accuracy and reducing the number of inconclusive results.Methodology/Principal findingsHere, four chimeric proteins (IBMP-8.1 to -8.4) comprising immunodominant regions of different Trypanosoma cruzi antigens were tested by enzyme-linked immunosorbent assay. The proteins were used to detect specific anti-T. cruzi antibodies in the sera of 857 chagasic and 689 non-chagasic individuals to evaluate their accuracy for chronic CD diagnosis. The antigens were recombinantly expressed in Escherichia coli and purified by chromatographic methods. The sensitivity and specificity values ranged from 94.3% to 99.3% and 99.4% to 100%, respectively. The diagnostic odds ratio (DOR) values were 6,462 for IBMP-8.1, 3,807 for IBMP-8.2, 32,095 for IBMP-8.3, and 283,714 for IBMP-8.4. These chimeric antigens presented DORs that were higher than the commercial test Pathozyme Chagas. The antigens IBMP-8.3 and -8.4 also showed DORs higher than the Gold ELISA Chagas test. Mixtures with equimolar concentrations were tested in order to improve the diagnosis accuracy of negative samples with high signal and positive samples with low signal. However, no gain in accuracy was observed relative to the individual antigens. A total of 1,079 additional sera were used to test cross-reactivity to unrelated diseases. The cross-reactivity rates ranged from 0.37% to 0.74% even for Leishmania spp., a pathogen showing relatively high genome sequence identity to T. cruzi. Imprecision analyses showed that IBMP chimeras are very stable and the results are highly reproducible.Conclusions/SignificanceOur findings indicate that the IBMP-8.4 antigen can be safely used in serological tests for T. cruzi screening in blood banks and for chronic CD laboratory diagnosis.

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Use of an enzyme-linked immunosorbent assay that utilizes the Tc13Tul antigen of Trypanosoma cruzi to monitor patients after treatment with benznidazole

Cited by 6 publications

References 41 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

A novel antibody surrogate biomarker to monitor parasite persistence in Trypanosoma cruzi-infected patients

Accuracy of chimeric proteins in the serological diagnosis of chronic chagas disease – a Phase II study

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