BACKGROUND: Canine leishmaniasis (CanL) is a severe chronic disease caused by Leishmania infantum and transmitted by sand flies of which the main vector in the Western part of the Mediterranean basin is Phlebotomus perniciosus. Previously, an immunochromatographic test (ICT) was proposed to allow rapid evaluation of dog exposure to P. perniciosus. In the present study, we optimized the prototype and evaluated the detection accuracy of the ICT in field conditions. Possible cross-reactions with other hematophagous arthropods were also assessed. METHODOLOGY/PRINCIPAL FINDINGS: The ICT was optimized by expressing the rSP03B protein in a HEK293 cell line, which delivered an increased specificity (94.92%). The ICT showed an excellent reproducibility and inter-person reliability, and was optimized for use with whole canine blood which rendered an excellent degree of agreement with the use of serum. Field detectability of the ICT was assessed by screening 186 dogs from different CanL endemic areas with both the SGH-ELISA and the ICT, and 154 longitudinally sampled dogs only with the ICT. The ICT results corresponded to the SGH-ELISA for most areas, depending on the statistical measure used. Furthermore, the ICT was able to show a clear seasonal fluctuation in the proportion of bitten dogs. Finally, we excluded cross-reactions between non-vector species and confirmed favorable cross-reactions with other L. infantum vectors belonging to the subgenus Larroussius. CONCLUSIONS/SIGNIFICANCE: We have successfully optimized the ICT, now also suitable to be used with whole canine blood. The test is able to reflect the seasonal fluctuation in dog exposure and showed a good detectability in a field population of naturally exposed dogs, particularly in areas with a high seroprevalence of bitten dogs. Furthermore, our study showed the existence of favorable cross-reactions with other sand fly vectors thereby expanding its use in the field.

Coris BioConcept Crealys Park Gembloux Belgium

Department of Biochemistry Faculty of Science Charles University Prague Czech Republic

Department of Parasitology Faculty of Science Charles University Prague Czech Republic

Hospital Veterinari Canis Girona Spain

ISGlobal Hospital Clínic Universitat de Barcelona Barcelona Spain

Secció de Parasitologia Departament de Biologia Sanitat i Medi Ambient Facultat de Farmàcia i Ciències de l'Alimentació Universitat de Barcelona Barcelona Spain

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Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 2012;7: e35671 10.1371/journal.pone.0035671 PubMed DOI PMC

Killick-Kendrick R, Killick-Kendrick M. Biology of sand fly vectors of Mediterranean canine leishmaniasis In: Killick-Kendrick R, editor. Canine Leishmaniasis: an update Proceedings of the International Canine Leishmaniasis Forum. Hoechst Roussel Vet; 1999. pp. 26–31.

Baneth G, Koutinas AF, Solano-Gallego L, Bourdeau P, Ferrer L. Canine leishmaniosis—new concepts and insights on an expanding zoonosis: part one. Trends Parasitol. 2008;24: 324–330. 10.1016/j.pt.2008.04.001 PubMed DOI

Molina R, Amela C, Nieto J, Gonzalez F, Castillo JA, Lucientes J, et al. Infectivity of dogs naturally infected with PubMed DOI

Miró G, Checa R, Montoya A, Hernández L, Dado D, Gálvez R. Current situation of PubMed DOI PMC

Moreno J, Alvar J. Canine leishmaniasis: Epidemiological risk and the experimental model. Trends Parasitol. 2002;18: 399–405. 10.1016/s1471-4922(02)02347-4 PubMed DOI

Ferroglio E, Maroli M, Gastaldo S, Mignone W, Rossi L. Canine leishmaniasis, Italy. Emerg Infect Dis. 2005;11: 1618–1620. 10.3201/eid1110.040966 PubMed DOI PMC

Maresca C, Scoccia E, Barizzone F, Catalano A, Mancini S, Pagliacci T, et al. A survey on canine leishmaniasis and phlebotomine sand flies in central Italy. Res Vet Sci. Elsevier Ltd; 2009;87: 36–38. 10.1016/j.rvsc.2008.11.009 PubMed DOI

Solano-Gallego L, Morell P, Arboix M, Alberola J, Ferrer L. Prevalence of PubMed DOI PMC

Berrahal F, Mary C, Roze M, Berenger A, Escoffier K, Lamouroux D, et al. Canine leishmaniasis: Identification of asymptomatic carriers by polymerase chain reaction and immunoblotting. Am J Trop Med Hyg. 1996;55: 273–277. 10.4269/ajtmh.1996.55.273 PubMed DOI

Otranto D, Dantas-Torres F. The prevention of canine leishmaniasis and its impact on public health. Trends Parasitol. Elsevier Ltd; 2013;29: 339–345. 10.1016/j.pt.2013.05.003 PubMed DOI

Vlkova M, Rohousova I, Hostomska J, Pohankova L, Zidkova L, Drahota J, et al. Kinetics of antibody response in BALB/c and C57BL/6 mice bitten by PubMed DOI PMC

Vlkova M, Rohousova I, Drahota J, Stanneck D, Kruedewagen EM, Mencke N, et al. Canine antibody response to PubMed DOI PMC

Kostalova T, Lestinova T, Sumova P, Vlkova M, Rohousova I, Berriatua E, et al. Canine Antibodies against Salivary Recombinant Proteins of PubMed DOI PMC

Sima M, Ferencova B, Warburg A, Rohousova I, Volf P. Recombinant Salivary Proteins of PubMed DOI PMC

Teixeira C, Gomes R, Collin N, Reynoso D, Jochim R, Oliveira F, et al. Discovery of markers of exposure specific to bites of PubMed DOI PMC

Souza AP, Andrade BB, Aquino D, Entringer P, Miranda JC, Alcantara R, et al. Using recombinant proteins from PubMed DOI PMC

Kostalova T, Lestinova T, Maia C, Sumova P, Vlkova M, Willen L, et al. The recombinant protein rSP03B is a valid antigen for screening dog exposure to PubMed DOI

Velez R, Spitzova T, Domenech E, Willen L, Cairó J, Volf P, et al. Seasonal dynamics of canine antibody response to PubMed DOI PMC

Willen L, Mertens P, Volf P. Evaluation of the rSP03B sero-strip, a newly proposed rapid test for canine exposure to PubMed DOI PMC

Volf P, Volfova V. Establishment and maintenance of sand fly colonies. J Vector Ecol. 2011;36: S1–S9. 10.1111/j.1948-7134.2011.00106.x PubMed DOI

Durocher Y. High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res. 2002;30: e9 10.1093/nar/30.2.e9 PubMed DOI PMC

Bláha J, Pachl P, Novák P, Vaněk O. Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI- cells. Protein Expr Purif. 2015;109: 7–13. 10.1016/j.pep.2015.01.006 PubMed DOI

R Core Team (2015). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria: [Internet]. Available: https://www.r-project.org/

Wickham H. ggplot2: Elegant Graphics for Data Analysis Springer-Verlag; New York, 2009.

Cohen J. A coefficient of agreement for nominal scales. Educ Psychol Meas. 1960;20: 37–46.

McGraw KO, Wong SP. Forming Inferences about Some Intraclass Correlation Coefficients. Psychol Methods. 1996;1: 30–46. 10.1037/1082-989X.1.1.30 DOI

Cicchetti D V. Guidelines, Criteria, and Rules of Thumb for Evaluating Normed and Standardized Assessment Instruments in Psychology. Psychol Assess. 1994;6: 284–290. 10.1037/1040-3590.6.4.284 DOI

Sajid M, Kawde AN, Daud M. Designs, formats and applications of lateral flow assay: A literature review. J Saudi Chem Soc. 2015;19: 689–705. 10.1016/j.jscs.2014.09.001 DOI

Lestinova T, Vlkova M, Votypka J, Volf P, Rohousova I. PubMed DOI

Volf P, Rohoušová I. Species-specific antigens in salivary glands of phlebotomine sandflies. Parasitology. 2001;122: 37–41. 10.1017/s0031182000007046 PubMed DOI

Lawyer P, Killick-Kendrick M, Rowland T, Rowton E, Volf P. Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae). Parasite. 2017;24: 42 10.1051/parasite/2017041 PubMed DOI PMC

Stockholm: ECDC. European Centre for Disease Prevention and Control and European Food Safety Authority. Phlebotomine sandflies maps [Internet]. 2018. Available: https://ecdc.europa.eu/en/disease-vectors/surveillance-and-disease-data/phlebotomine-maps

Alten B, Maia C, Afonso MO, Campino L, Jiménez M, González E, et al. Seasonal Dynamics of Phlebotomine Sand Fly Species Proven Vectors of Mediterranean Leishmaniasis Caused by PubMed DOI PMC

Kamhawi S. Phlebotomine sand flies and PubMed DOI

Volf P, Peckova J. Sand flies and PubMed DOI PMC

Humoral and T-Cell-Mediated Immunity Against Phlebotomus perniciosus Salivary Proteins in Dogs from a Leishmaniosis-Endemic Area

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