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. 2016 Dec 9;11(12):e0168074.
doi: 10.1371/journal.pone.0168074. eCollection 2016.

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

Sylvain Biéler  1 , Harald Waltenberger  2 , Michael P Barrett  3 , Richard McCulloch  3 , Jeremy C Mottram  3 , Mark Carrington  4 , Wilhelm Schwaeble  5 , James McKerrow  6 , Margaret A Phillips  7 , Paul A Michels  8 , Philippe Büscher  9 , Jean-Charles Sanchez  10 , Richard Bishop  11 , Derrick R Robinson  12 , James Bangs  13 , Michael Ferguson  14 , Barbara Nerima  15 , Audrey Albertini  1 , Gerd Michel  1 , Magdalena Radwandska  1 , Joseph Mathu Ndung'u  1
Affiliations

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

Sylvain Biéler et al. PLoS One. .

Abstract

Background: Control 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 findings: The 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.

Conclusions: This 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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Conflict of interest statement

HW’s affiliation to MicroCoat Biotechnologie GmbH does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1
Percentage IgG (A) and IgM (B) reactivity of 32 antigens on sera from T. b. gambiense HAT cases (N = 40) assessed by slot blot (first round of screening). Antigens are shown in descending order of reactivity. Antigens marked with an asterisk (*) were only partially soluble.
Fig 2
Fig 2
Percentage IgG (A) and IgM (B) reactivity of 32 antigens on sera from T. b. rhodesiense HAT cases (N = 10) assessed by slot blot (first round of screening). Antigens are shown in descending order of reactivity. Antigens marked with an asterisk (*) were only partially soluble.
Fig 3
Fig 3
Percentage IgG reactivity of 18 antigens on sera from T. b. gambiense HAT cases (N = 35) assessed by slot blot (A) and by ELISA (B) (second round of screening). Antigens are shown in descending order of reactivity.
Fig 4
Fig 4
Percentage IgG reactivity of 18 antigens on sera from T. b. rhodesiense HAT cases (N = 10) assessed by slot blot (A) and by ELISA (B) (second round of screening). Antigens are shown in descending order of reactivity.
Fig 5
Fig 5
Percentage IgG reactivity of 13 antigens on sera from T. b. gambiense HAT cases (N = 150) assessed by slot blot (A) and by ELISA (B) (third round of screening). Antigens are shown in descending order of reactivity.
Fig 6
Fig 6
Percentage IgG reactivity of 13 antigens on sera from T. b. rhodesiense HAT cases (N = 33) assessed by slot blot (A) and by ELISA (B) (third round of screening). Antigens are shown in descending order of reactivity.
See this image and copyright information in PMC

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