Proteomic Selection of Immunodiagnostic Antigens for Human African Trypanosomiasis and Generation of a Prototype Lateral Flow Immunodiagnostic Device

Sullivan, Louis W.; Wall, Stephen; Carrington, Mark; Ferguson, Michael A. J.

doi:10.1371/journal.pntd.0002087

Cited by 53 publications

(71 citation statements)

References 49 publications

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“…Unlike human African trypanosomosis, where a lateral flow device prototype has recently been developed, no point-of-treatment test exists for AAT [6,7]. The current gold standard for diagnosis of AAT is examination of blood by light microscopy for the presence of parasites.…”

Section: Introductionmentioning

confidence: 99%

Trypanosoma vivax GM6 Antigen: A Candidate Antigen for Diagnosis of African Animal Trypanosomosis in Cattle

et al. 2013

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BackgroundDiagnosis of African animal trypanosomosis is vital to controlling this severe disease which hampers development across 10 million km2 of Africa endemic to tsetse flies. Diagnosis at the point of treatment is currently dependent on parasite detection which is unreliable, and on clinical signs, which are common to several other prevalent bovine diseases.Methodology/Principle Findingsthe repeat sequence of the GM6 antigen of Trypanosoma vivax (TvGM6), a flagellar-associated protein, was analysed from several isolates of T. vivax and found to be almost identical despite the fact that T. vivax is known to have high genetic variation. The TvGM6 repeat was recombinantly expressed in E. coli and purified. An indirect ELISA for bovine sera based on this antigen was developed. The TvGM6 indirect ELISA had a sensitivity of 91.4% (95% CI: 91.3 to 91.6) in the period following 10 days post experimental infection with T. vivax, which decreased ten-fold to 9.1% (95% CI: 7.3 to 10.9) one month post treatment. With field sera from cattle infected with T. vivax from two locations in East and West Africa, 91.5% (95% CI: 83.2 to 99.5) sensitivity and 91.3% (95% CI: 78.9 to 93.1) specificity was obtained for the TvGM6 ELISA using the whole trypanosome lysate ELISA as a reference. For heterologous T. congolense field infections, the TvGM6 ELISA had a sensitivity of 85.1% (95% CI: 76.8 to 94.4).Conclusion/Significancethis study is the first to analyse the GM6 antigen of T. vivax and the first to test the GM6 antigen on a large collection of sera from experimentally and naturally infected cattle. This study demonstrates that the TvGM6 is an excellent candidate antigen for the development of a point-of-treatment test for diagnosis of T. vivax, and to a lesser extent T. congolense, African animal trypanosomosis in cattle.

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Section: Introductionmentioning

confidence: 99%

Trypanosoma vivax GM6 Antigen: A Candidate Antigen for Diagnosis of African Animal Trypanosomosis in Cattle

et al. 2013

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“…The identified proteins (Table S1 in Text S1) are mostly surface glycoproteins such as variant surface glycoprotein B (VSG_B), expression site associated genes (ESAGs), invariant surface glycoproteins (ISGs), trans-sialidases, unknown proteins that have signal peptides and/or transmembrane domains and proteins involved in trafficking such as soluble NSF attachment protein receptors (SNAREs). These surface and secretory pathway glycoproteins are similar to the proteins identified in a comparable study using human Trypanosoma brucei gambiense infection and control IgGs [7]. Fifteen of the identified T. congolense proteins were selected for expression trials based on assessment for predicted ease of expression in E. coli .…”

Section: Resultsmentioning

confidence: 79%

“…This was achieved by following previously established methods [7]. Briefly, IgG from pooled pre- and 28 day post-infected cattle sera were immobilised onto Sepharose beads and incubated with T. congolense lysate.…”

Section: Resultsmentioning

confidence: 99%

Proteomic Selection of Immunodiagnostic Antigens for Trypanosoma congolense

et al. 2014

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Animal African Trypanosomosis (AAT) presents a severe problem for agricultural development in sub-Saharan Africa. It is caused by several trypanosome species and current means of diagnosis are expensive and impractical for field use. Our aim was to discover antigens for the detection of antibodies to Trypanosoma congolense, one of the main causative agents of AAT. We took a proteomic approach to identify potential immunodiagnostic parasite protein antigens. One hundred and thirteen proteins were identified which were selectively recognized by infected cattle sera. These were assessed for likelihood of recombinant protein expression in E. coli and fifteen were successfully expressed and assessed for their immunodiagnostic potential by ELISA using pooled pre- and post-infection cattle sera. Three proteins, members of the invariant surface glycoprotein (ISG) family, performed favorably and were then assessed using individual cattle sera. One antigen, Tc38630, evaluated blind with 77 randomized cattle sera in an ELISA assay gave sensitivity and specificity performances of 87.2% and 97.4%, respectively. Cattle immunoreactivity to this antigen diminished significantly following drug-cure, a feature helpful for monitoring the efficacy of drug treatment.

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“…It is possible that other antigens that were either not included or had not been identified at that time could perform equally well or better than assessed in the present study. For example, the flagellar calcium-binding protein TB-17 [20,60], the Gene Related to Expression Site Associated Gene (GRESAG) 4, and transferrin receptor subunits ESAG 6 and 7 have also been suggested as potential candidates [20]. Since the most reactive antigens include the native VSG LiTat 1.3 and VSG LiTat 1.5 antigens, which are costly and difficult to produce, one approach might be to generate recombinant versions of these proteins.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Antigens for Development of a Serological Test for Human African Trypanosomiasis

Biéler

Waltenberger²,

Barrett

et al. 2016

PLoS ONE

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BackgroundControl and elimination of human African trypanosomiasis (HAT) can be accelerated through the use of diagnostic tests that are more accurate and easier to deploy. The goal of this work was to evaluate the immuno-reactivity of antigens and identify candidates to be considered for development of a simple serological test for the detection of Trypanosoma brucei gambiense or T. b. rhodesiense infections, ideally both.Methodology/Principal FindingsThe reactivity of 35 antigens was independently evaluated by slot blot and ELISA against sera from both T. b. gambiense and T. b. rhodesiense infected patients and controls. The antigens that were most reactive by both tests to T. b. gambiense sera were the membrane proteins VSG LiTat 1.3, VSG LiTat 1.5 and ISG64. Reactivity to T. b. rhodesiense sera was highest with VSG LiTat 1.3, VSG LiTat 1.5 and SRA, although much lower than with T. b. gambiense samples. The reactivity of all possible combinations of antigens was also calculated. When the slot blot results of 2 antigens were paired, a VSG LiTat 1.3- ISG75 combination performed best on T. b. gambiense sera, while a VSG LiTat 1.3-VSG LiTat 1.5 combination was the most reactive using ELISA. A combination of SRA and either VSG LiTat 1.3 or VSG LiTat 1.5 had the highest reactivity on T. b. rhodesiense sera according to slot blot, while in ELISA, pairing SRA with either GM6 or VSG LiTat 1.3 yielded the best results.ConclusionsThis study identified antigens that were highly reactive to T. b. gambiense sera, which could be considered for developing a serological test for gambiense HAT, either individually or in combination. Antigens with potential for inclusion in a test for T. b. rhodesiense HAT were also identified, but because their reactivity was comparatively lower, a search for additional antigens would be required before developing a test for this form of the disease.

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Proteomic Selection of Immunodiagnostic Antigens for Human African Trypanosomiasis and Generation of a Prototype Lateral Flow Immunodiagnostic Device

Cited by 53 publications

References 49 publications

Trypanosoma vivax GM6 Antigen: A Candidate Antigen for Diagnosis of African Animal Trypanosomosis in Cattle

Trypanosoma vivax GM6 Antigen: A Candidate Antigen for Diagnosis of African Animal Trypanosomosis in Cattle

Proteomic Selection of Immunodiagnostic Antigens for Trypanosoma congolense

Evaluation of Antigens for Development of a Serological Test for Human African Trypanosomiasis

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