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Description of a nanobody-based competitive immunoassay to detect tsetse fly exposure

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Abstract
Background : Tsetse flies are the main vectors of human and animal African trypanosomes. The Tsal proteins in tsetse fly saliva were previously identified as suitable biomarkers of bite exposure. A new competitive assay was conceived based on nanobody (Nb) technology to ameliorate the detection of anti-Tsal antibodies in mammalian hosts. Methodology/Principal Findings : A camelid-derived Nb library was generated against the Glossina morsitans morsitans sialome and exploited to select Tsal specific Nbs. One of the three identified Nb families (family III, TsalNb-05 and TsalNb-11) was found suitable for anti-Tsal antibody detection in a competitive ELISA format. The competitive ELISA was able to detect exposure to a broad range of tsetse species (G. morsitans morsitans, G. pallidipes, G. palpalis gambiensis and G. fuscipes) and did not cross-react with the other hematophagous insects (Stomoxys calcitrans and Tabanus yao). Using a collection of plasmas from tsetse-exposed pigs, the new test characteristics were compared with those of the previously described G. m. moristans and rTsal1 indirect ELISAs, revealing equally good specificities (> 95%) and positive predictive values (> 98%) but higher negative predictive values and hence increased sensitivity (> 95%) and accuracy (> 95%). Conclusion/Significance : We have developed a highly accurate Nb-based competitive immunoassay to detect specific anti-Tsal antibodies induced by various tsetse fly species in a range of hosts. We propose that this competitive assay provides a simple serological indicator of tsetse fly presence without the requirement of test adaptation to the vertebrate host species. In addition, the use of monoclonal Nbs for antibody detection is innovative and could be applied to other tsetse fly salivary biomarkers in order to achieve a multi-target immunoprofiling of hosts. In addition, this approach could be broadened to other pathogenic organisms for which accurate serological diagnosis remains a bottleneck.
Keywords
IGG ANTIBODY-RESPONSE, HUMAN AFRICAN TRYPANOSOMIASIS, WEST-AFRICA, VISCERAL LEISHMANIASIS, SALIVARY ANTIGENS, IMMUNE-RESPONSE, BITES, IDENTIFICATION, BIOMARKER, VECTOR

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Chicago
Caljon, Guy, Shahid Hussain, Lieve Vermeiren, and Jan Van Den Abbeele. 2015. “Description of a Nanobody-based Competitive Immunoassay to Detect Tsetse Fly Exposure.” Plos Neglected Tropical Diseases 9 (2).
APA
Caljon, Guy, Hussain, S., Vermeiren, L., & Van Den Abbeele, J. (2015). Description of a nanobody-based competitive immunoassay to detect tsetse fly exposure. PLOS NEGLECTED TROPICAL DISEASES, 9(2).
Vancouver
1.
Caljon G, Hussain S, Vermeiren L, Van Den Abbeele J. Description of a nanobody-based competitive immunoassay to detect tsetse fly exposure. PLOS NEGLECTED TROPICAL DISEASES. 2015;9(2).
MLA
Caljon, Guy et al. “Description of a Nanobody-based Competitive Immunoassay to Detect Tsetse Fly Exposure.” PLOS NEGLECTED TROPICAL DISEASES 9.2 (2015): n. pag. Print.
@article{6905883,
  abstract     = {Background : Tsetse flies are the main vectors of human and animal African trypanosomes. The Tsal proteins in tsetse fly saliva were previously identified as suitable biomarkers of bite exposure. A new competitive assay was conceived based on nanobody (Nb) technology to ameliorate the detection of anti-Tsal antibodies in mammalian hosts. 
Methodology/Principal Findings : A camelid-derived Nb library was generated against the Glossina morsitans morsitans sialome and exploited to select Tsal specific Nbs. One of the three identified Nb families (family III, TsalNb-05 and TsalNb-11) was found suitable for anti-Tsal antibody detection in a competitive ELISA format. The competitive ELISA was able to detect exposure to a broad range of tsetse species (G. morsitans morsitans, G. pallidipes, G. palpalis gambiensis and G. fuscipes) and did not cross-react with the other hematophagous insects (Stomoxys calcitrans and Tabanus yao). Using a collection of plasmas from tsetse-exposed pigs, the new test characteristics were compared with those of the previously described G. m. moristans and rTsal1 indirect ELISAs, revealing equally good specificities (> 95%) and positive predictive values (> 98%) but higher negative predictive values and hence increased sensitivity (> 95%) and accuracy (> 95%). 
Conclusion/Significance : We have developed a highly accurate Nb-based competitive immunoassay to detect specific anti-Tsal antibodies induced by various tsetse fly species in a range of hosts. We propose that this competitive assay provides a simple serological indicator of tsetse fly presence without the requirement of test adaptation to the vertebrate host species. In addition, the use of monoclonal Nbs for antibody detection is innovative and could be applied to other tsetse fly salivary biomarkers in order to achieve a multi-target immunoprofiling of hosts. In addition, this approach could be broadened to other pathogenic organisms for which accurate serological diagnosis remains a bottleneck.},
  articleno    = {e0003456},
  author       = {Caljon, Guy and Hussain, Shahid and Vermeiren, Lieve and Van Den Abbeele, Jan},
  issn         = {1935-2735},
  journal      = {PLOS NEGLECTED TROPICAL DISEASES},
  keywords     = {IGG ANTIBODY-RESPONSE,HUMAN AFRICAN TRYPANOSOMIASIS,WEST-AFRICA,VISCERAL LEISHMANIASIS,SALIVARY ANTIGENS,IMMUNE-RESPONSE,BITES,IDENTIFICATION,BIOMARKER,VECTOR},
  language     = {eng},
  number       = {2},
  pages        = {18},
  title        = {Description of a nanobody-based competitive immunoassay to detect tsetse fly exposure},
  url          = {http://dx.doi.org/10.1371/journal.pntd.0003456},
  volume       = {9},
  year         = {2015},
}

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