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A versatile and sensitive lateral flow immunoassay for the rapid diagnosis of visceral leishmaniasis

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Abstract

Visceral leishmaniasis (VL) is a zoonotic infectious disease with a severe impact on humans and animals. Infection is transmitted by phlebotomine sand flies, and several domestic and wild mammals act as reservoirs for the infection, so the prompt detection of infected hosts is crucial to preventing and controlling the spread of the disease and its transmission to humans. A rapid and portable tool for VL diagnosis based on the lateral flow immunoassay (LFIA) technology is described herein. The device exploits a highly specific chimeric recombinant antigen as the recognition element for capturing anti-leishmanial antibodies, and protein A labelled with gold nanoparticles as the signal reporter. The LFIA shows excellent diagnostic sensitivity (98.4%), specificity (98.9%), and agreement with serological reference methods for diagnosing canine VL. The long-term stability of the LFIA device was confirmed based on six months of storage at room temperature or 4 °C, and the qualitative response of the device was not affected by limited thermal stress. The use of the broadly specific protein A means that the LFIA can be readily adapted to diagnose VL in dogs (the main reservoir for human infection) and other mammals, thus further assisting efforts to control the spread of VL.

A rapid and portable diagnostic tool for visceral leishmaniasis (VL) based on lateral flow immunoassay (LFIA) technology. The presence of anti-leishmanial antibodies is revealed through the binding of these antibodies to a highly specific chimeric antigen. Employing a broadly specific signal reporter (protein A labelled with gold nanoparticles) enables the LFIA to be easily adapted to diagnose VL in different animals

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References

  1. Solca MDS, Bastos LA, Guedes CES, Bordoni M, Borja LS, et al. Evaluating the accuracy of molecular diagnostic testing for canine visceral leishmaniasis using latent class analysis. PLoS ONE. 2014;9(7):e103635. https://doi.org/10.1371/journal.pone.0103635.

    Article  CAS  PubMed Central  Google Scholar 

  2. Adams E, Hasker E, Cunningham J. Visceral leishmaniasis rapid diagnostic test performance. Diagnostic evaluation series no. 4. Geneva: World Health Organization; 2011.

  3. Adel A, Berkvens D, Abatih E, Soukehal A, Bianchini J, Saegerman C. Evaluation of immunofluorescence antibody test used for the diagnosis of canine leishmaniasis in the Mediterranean Basin: a systematic review and meta-analysis. PLoS One. 2016;11(8):e0161051. https://doi.org/10.1371/journal.pone.0161051.

  4. Ferroglio E, Maroli M, Gastaldo S, Mignone M, Rossi L. Canine leishmaniosis in Italy. Emerg Infect Dis. 2005;11:1618–20.

    Article  PubMed  PubMed Central  Google Scholar 

  5. World Health Organization. Media centre: Leishmaniasis.http://www.who.int/mediacentre/factsheets/fs375/en/. Accessed 23 Jan 2018.

  6. Woyames Pinto AJ, Ribeiro VM, Tafuri WL. The immunochromatography use in canine visceral leishmaniasis in Brazil: a “quick solution” of a complex diagnostic? Rapid test in dogs with leishmaniasis. Ann Clin Cytol Pathol. 2016;2(4):1033.

  7. The World Organization for Animal Health. Leishmaniasis. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.01.08_LEISHMANIOSIS.pdf. Accessed 22 Jan 18.

  8. Paltrinieri S, Gradoni L, Roura X, Zatelli A, Zini E. Laboratory tests for diagnosing and monitoring canine leishmaniasis. Vet Clin Pathol. 2016;45:552–78.

    Article  PubMed  Google Scholar 

  9. St John A, Price CP. Existing and emerging technologies for point-of-care testing. Clin Biochem Rev. 2014;35:155–67.

    PubMed  PubMed Central  Google Scholar 

  10. IDEXX Laboratories. SNAP Leishmania Test.https://www.idexx.eu/en/products-and-solutions/SNAP-and-Pet-Side-Tests/snap-leishmania-test/. Accessed 23 Jan 2018.

  11. InBios International, Inc. Kalazar Detect™ Rapid Test for Visceral Leishmaniasis.http://www.inbios.com/kalazar-detecttm-rapid-test-for-visceral-leishmaniasis-intl/ Accessed 23 Jan 2018.

  12. Chembio Diagnostic Systems, Inc. Dual Path Platform.http://chembio.com/innovation/platforms/dual-path-platform/. Accessed 23 Jan 2018.

  13. Virbac BVT. Speed Leish K. https://bvt.virbac.com/en/home/diagnostic-solutions/pour-le-veterinaire-praticien/vector-borne-and-parasitic-disea/main/produits/speed-leish-k.html. Accessed 23 Jan 2018.

  14. Martins Cardoso R, Silva Lociks de Araújo NN, Sierra Romero GA, Castro Minuzzi Souza TT, Dietrich AG, Donizette Mendes J, et al. Expanding the knowledge about Leishmania species in wild mammals and dogs in the Brazilian savannah. Parasit Vectors. 2015;8:171. https://doi.org/10.1186/s13071-015-0780-y.

  15. Gao C-H, Wang J-Y, Zhang S, Yang Y-T, Wang Y. Survey of wild and domestic mammals for infection with Leishmania infantum following an outbreak of desert zoonotic visceral leishmaniasis in Jiashi, People's Republic of China. PLoS One. 2015;10(7):e0132493. https://doi.org/10.1371/journal.pone.0132493.

  16. Rosati S, Ortoffi M, Profiti M, Mannelli A, Mignone W, Bollo E, et al. Prokaryotic expression and antigenic characterization of three recombinant Leishmania antigens for serological diagnosis of canine leishmaniasis. Clin Diagn Lab Immunol. 2003;10:1153–6. https://doi.org/10.1128/CDLI.10.6.1153-1156.2003.

  17. Boarino A, Scalone A, Gradoni L, Ferroglio E, Vitale F, Zanatta R, et al. Development of recombinant chimeric antigen expressing immunodominant B epitopes of Leishmania infantum for serodiagnosis of visceral leishmaniasis. Diag Lab Immunol. 2005;5:647–53.

  18. Stobel K, Schonberg A, Staak C. A new non-species dependent ELISA for detection of antibodies to Borrelia burgdorferi s. I. in zoo animals. Int J Med Microbiol. 2002;291(33):88–99.

  19. Al-Adhami BH, Gajadhar AA. A new multi-host species indirect ELISA using protein A/G conjugate for detection of anti-Toxoplasma gondii IgG antibodies with comparison to ELISA-IgG, agglutination assay and Western blot. Vet Parasitol. 2014;200:66–73. https://doi.org/10.1016/j.vetpar.2013.11.004.

  20. Scalone DF, Mignone W, Ferroglio E, Mannelli A, Biglino A, Zanatta R, et al. Validation of a recombinant based antibody ELISA for diagnosis of human and canine leishmaniasis. J Immun Immunochem. 2008;29:244–56. https://doi.org/10.1080/15321810802116006.

  21. Intaramat A, Sornprachum T, Chantrathonkul B, Chaisuriya P, Lohnoo T, Yingyong W, et al. Protein A/G-based immunochromatographic test for serodiagnosis of pythiosis in human and animal subjects from Asia and Americas. Med Mycol Open Access. 2016;54:641–7. https://doi.org/10.1093/mmy/myw018.

  22. Omachi S, Matsumoto Y, Goto Y. Immunoglobulins in the pathophysiology of visceral leishmaniasis. In: Noiri E, Jha T, editors. Kala azar in South Asia. Cham: Springer; 2016.

  23. Anfossi L, Calderara M, Baggiani C, Giovannoli C, Arletti E, Giraudi G. Development and application of a quantitative lateral flow immunoassay for fumonisins in maize. Anal Chim Acta. 2010;682:104–9. https://doi.org/10.1016/j.aca.2010.09.045.

    Article  CAS  PubMed  Google Scholar 

  24. Ferroglio E, Zanet S, Mignone W, Poggi M, Trisciuoglio A, Bianciardi P. Evaluation of a rapid device for serological diagnosis of Leishmania infantum infection in dog, compared to immunofluorescence assay and western blot. Clin Vaccine Immunol. 2013;20:657–9.

  25. Lattanzio VMT, von Holst C, Visconti A. Experimental design for in-house validation of a screening immunoassay kit. The case of a multiplex dipstick for Fusarium mycotoxins in cereals. Anal Bioanal Chem. 2013;405:7773–82.

  26. Lattanzio VMT, Guarducci N, Powers S, Ciasca B, Pascale M, von Holst C. Validation of a lateral flow immunoassay for the rapid determination of aflatoxins in maize by solvent free extraction. Anal Methods. 2018;10:123–30. https://doi.org/10.1039/C7AY02249B.

    Article  CAS  Google Scholar 

  27. Di Nardo F, Anfossi L, Ozella L, Saccani A, Giovannoli C, Spano G, et al. Validation of a qualitative immunochromatographic test for the noninvasive assessment of stress in dogs. J Chromatog B. 2016;1028:192–8. https://doi.org/10.1016/j.jchromb.2016.06.019.

  28. Kaplan LA, Pesce AJ. Clinical chemistry. Theory, analysis, correlation. 5th ed. Maryland Heights, MO: Mosby Elsevier Inc.; 2010.

  29. Da Silva ES, Van der Meide WF, Schoone GJ, Gontijo CMF, Schallig HDFH, Brazil RP. Diagnosis of canine leishmaniasis in the endemic area of Belo Horizonte, Minas Gerais, Brazil by parasite, antibody and DNA detection assays. Vet Res Comm. 2006;30:637–43. https://doi.org/10.1007/s11259-006-3324-2.

  30. Chun P. Colloidal gold and other labels for lateral flow immunoassays. In: Wong RC, Tse HY, editors. Lateral flow immunoassay. 1st ed. New York: Humana Press; 2009. p. 75–93.

    Google Scholar 

  31. Byzova NA, Safenkova IV, Slutskaya ES, Zherdev AV, Dzantiev BB. Less is more: a comparison of antibody-gold nanoparticle conjugates of different ratios. Bioconjug Chem. 2017;28:2737–46. https://doi.org/10.1021/acs.bioconjchem.7b00489.

    Article  CAS  PubMed  Google Scholar 

  32. Solano-Gallego L, Villanueva-Saz S, Carbonell M, Trotta M, Furlanello T, Natale A. Serological diagnosis of canine leishmaniosis: comparison of three commercial ELISA tests (Leiscan®, ID Screen® and Leishmania 96®), a rapid test (Speed Leish K®) and an in-house IFAT. Parasit Vectors. 2014;7:111. https://doi.org/10.1186/1756-3305-7-111.

  33. Moreira OC, Yadon ZE, Cupolillo E. The applicability of real-time PCR in the diagnostic of cutaneous leishmaniasis and parasite quantification for clinical management: current status and perspectives. Acta Trop. 2017, in press; https://doi.org/10.1016/j.actatropica.2017.09.020.

  34. Pomares C, Despierres L, Del Giudice P, Delaunay P, Michel G, Ferrua B, et al. Western blot analysis as an aid for the diagnosis of cutaneous leishmaniasis due to Leishmania major. Trans R Soc Trop Med Hyg. 2012;106:452–4. https://doi.org/10.1016/j.trstmh.2012.03.001.

  35. Ferroglio E, Centaro E, Mignone W, Trisciuoglio A. Evaluation of an ELISA rapid device for the serological diagnosis of Leishmania infantum infection in dog as compared with immunofluorescence assay and western blot. Vet Parasitol. 2007;144:162–6.

  36. Otranto D, Paradies P, Sasanelli M, Leone N, De Caprariis D, Chirico J, et al. Recombinant K39 dipstick immunochromatographic test: a new tool for the serodiagnosis of canine leishmaniasis. J Vet Diagn Investig. 2005;17:32–7.

  37. Pennisi MG. Leishmaniosis of companion animals in Europe: an update vet. Parasitology. 2015;208:35–47.

    Google Scholar 

  38. Millán J, Ferroglio E, Solano-Gallego L. Role of wildlife in the epidemiology of Leishmania infantum infection in Europe. Parasitol Res. 2014;113:2005–14.

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Authors and Affiliations

  1. Department of Chemistry, University of Turin, Via Giuria 5, 10125, Turin, Italy

    Laura Anfossi, Fabio Di Nardo, Simone Cavalera, Claudio Baggiani, Cristina Giovannoli & Giulia Spano

  2. Department of Veterinary Science, University of Turin, Via Verdi 8, 10124, Turin, Italy

    Margherita Profiti, Ezio Ferroglio & Sergio Rosati

  3. In3Diagnostic s.r.l., Largo P. Braccini 2, 10095, Grugliasco, TO, Italy

    Chiara Nogarol

  4. Sez. Imperia, Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle D’Aosta, Via Bologna 148, 10154, Turin, Italy

    Walter Mignone

Authors
  1. Laura Anfossi
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  2. Fabio Di Nardo
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Correspondence to Laura Anfossi.

Ethics declarations

Disclosure of potential conflicts of interest

The author Chiara Nogarol works at the company In3Diagnostic (L.go P. Braccini 2, Grugliasco (TO), Italy), which may commercialise the LFIA device. However, this does not alter the authors’ adherence to the principles of good scientific practice and to relevant policies on sharing data and materials. The authors declare that they have no other competing interests.

Research involving animal participants

Blood samples were obtained during routine activities at the Veterinary Teaching Hospital of the Department of Veterinary Science or by veterinary practitioners. The study utilized samples from dogs at a private kennel/small-animal veterinary clinics, and informed consent was obtained from the owner of each dog. The consent was provided in oral form. No additional permission was required. All procedures were conducted in accordance with EU Directive 2010/63/EU for animal experiments, as well as with the informed consent of the owner of the animal.

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Anfossi, L., Di Nardo, F., Profiti, M. et al. A versatile and sensitive lateral flow immunoassay for the rapid diagnosis of visceral leishmaniasis. Anal Bioanal Chem 410, 4123–4134 (2018). https://doi.org/10.1007/s00216-018-1067-x

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  • DOI: https://doi.org/10.1007/s00216-018-1067-x

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