The rK39 Antigen from an Iranian Strain of Leishmania infantum: Detection of Anti-Leishmania Antibodies in Humans and Dogs

  • Bibi Razieh HOSSEINI FARASH Department of Medical Parasitology and Mycology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran . Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran .Cutaneous Leishmaniasis Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mehdi MOHEBALI Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran .Center for Research of Endemic Parasites of Iran, Tehran University of Medical Sciences, Tehran, Iran
  • Bahram KAZEMI Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Homa HAJJARAN Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Abdolmajid FATA Department of Medical Parasitology and Mycology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran .Cutaneous Leishmaniasis Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Reza RAOOFIAN Legal Medicine Research Center, Legal Medicine Organization, Tehran, Iran
  • Behnaz AKHOUNDI Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Majid MOJARRAD Department of Medical Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
  • Pietro MASTROENI Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom
  • Mohammad Kazem SHARIFI-YAZDI Center for Research of Zoonoses of Iran, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Hossein TANIPOUR Department of Medical Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
Keywords:
rK39 recombinant anti-gen, Leishmania infantum, Visceral leishmaniasis, Human, Dog

Abstract

Background: Visceral leishmaniasis (VL) is the most severe form of leishmaniasis in Iran with high mortality rates in the case of inaccurate diagnosis and treatment. This study aimed to prepare and evaluate a new rk39 recombinant antigen from an Iranian strain of Leishmania infantum for diagnosis of VL in humans and dogs. Methods: rK39-based enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for the detection of anti L. infantum antibodies. We screened 84 human sera and 87 dog sera from clinical cases in the endemic area of Meshkin-Shahr, Iran along with 176 sera from healthy controls (collected from 86 humans and 90 dogs) during 2013 -2016. Results: Using the rK39 ELISA, a sensitivity of 85.7% (95% CI, 95-99%) and a specificity of 86.0% (95% CI, 95%-99%) were detected in human sera at a 1:800 (cut-off) titer when DAT-confirmed cases were compared with healthy controls; a sensitivity of 96.6% (95% CI, 95%-99%) and specificity of 94.4% (95% CI, 95%-99%) were found at a 1:80 (cut-off) titer compared with DAT. Kappa analysis indicated agreement between the rK39 ELISA and DAT (0.718) when using human sera at a 1:800 (cut-off) titer as well as (0.910) at a 1:80 (cut-off) titer when using dog sera (P<0.05). Conclusion: New rk39 recombinant antigen from an Iranian strain of Leishmania infantum seems to be used for diagnosis of VL in humans and dogs. Further extended field studies are recommended.

References

1. Abass E, Kang C, Martinkovic F et al. Heterogenei-ty of Leishmania donovani Parasites Complicates Di-agnosis of Visceral Leishmaniasis: Comparison of Different Serological Tests in Three Endemic Re-gions. PLoS One. 2015; 10(3): e0116408.
2. Taran M, Mohebali M, Modaresi MH et al. Prepa-ration of a K39sub Recombinant Antigen for the Detection of Leishmania infantum Antibodies in Human: a Comparative Study with an Immuno-chromatographic Test and Direct Agglutination. Iran J Parasitol. 2007;2(2):25–33.
3. Savoia D. Recent updates and perspectives on leishmaniasis. J Infect Dev Ctries. 2015; 9(6):588–96.
4. Silva DT da, Starke-Buzetti WA, Alves-Martin MF et al. Comparative evaluation of several methods for Canine Visceral Leishmaniasis diagnosis. Rev Bras Parasitol Veterinária. 2014;23(2):179–86.
5. Sundar S, Rai M. Laboratory diagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol. 2002;9(5):951–8.
6. Taran M, Mohebali M, Modaresi MH et al. Diag-nosis of Canine Visceral Leishmaniasis by ELISA Using K39sub Recombinant Antigen. Iran J Public Health. 2007;36(2):1–6.
7. Farahmand M, Nahrevanian H. Application of Re-combinant Proteins for Serodiagnosis of Visceral Leishmaniasis in Humans and Dogs. Iran Biomed J. 2016;20(3):128–34.
8. Farajnia S, Darbani B, Babaei H et al. Development and evaluation of Leishmania infantum rK26 ELISA for serodiagnosis of visceral leishmaniasis in Iran. Parasitology. 2008;135(9):1035–41.
9. Mikaeili F, Fakhar M, Sarkari B et al. Comparison of serological methods (ELISA, DAT and IFA) for diagnosis of visceral leishmaniasis utilizing an en-demic strain. Iran J Immunol. 2007;4(2):116–21.
10. Mohebali M. Visceral leishmaniasis in Iran: Review of the Epidemiological and Clinical Features. Iran J Parasitol. 2013;8(3):348–58.
11. Badaró R, Benson D, Eulálio MC et al. rK39: a cloned antigen of Leishmania chagasi that predicts ac-tive visceral leishmaniasis. J Infect Dis. 1996;173(3):758–61.
12. Farash BRH, Mohebali M, Kazemi B et al. Cloning of K26 Hydrophilic Antigen from Iranian Strain of Leishmania infantum. Iran J Public Health. 2017; 46(10):1359–65.
13. Sakkas H, Gartzonika C, Levidiotou S. Laboratory diagnosis of human visceral leishmaniasis. J Vector Borne Dis. 2016;53(1):8–16.
14. Mohebali M, Taran M, Zarei Z. Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick rk39 test and direct agglutination. Vet Parasitol. 2004;121(3–4):239–45.
15. Mohebali M, Hajjaran H, Hamzavi Y et al. Epide-miological aspects of canine visceral leishmaniosis in the Islamic Republic of Iran. Vet Parasitol. 2005;129(3–4):243–51.
16. Le Rutte EA, Coffeng LE, Bontje DM et al. Feasi-bility of eliminating visceral leishmaniasis from the Indian subcontinent: explorations with a set of de-terministic age-structured transmission models. Parasit Vectors. 2016;9:24.
17. Stauch A, Sarkar RR, Picado A et al. Visceral Leishmaniasis in the Indian Subcontinent: Model-ling Epidemiology and Control. PLoS Negl Trop Dis. 2011;5(11):e1405.
18. Landis JR, Koch GG. The measurement of ob-server agreement for categorical data. Biometrics. 1977;33(1):159–74.
19. Sahdev S, Saini S, Tiwari P et al. Amplification of GC-rich genes by following a combination strategy of primer design, enhancers and modified PCR cy-cle conditions. Mol Cell Probes. 2007;21(4):303–7.
20. Siripattanapipong S, Kato H, Tan‐ariya P et al. Comparison of Recombinant Proteins of Kinesin 39, Heat Shock Protein 70, Heat Shock Protein 83, and Glycoprotein 63 for Antibody Detection of Leishmania martiniquensis Infection. J Eukaryot Microbiol. 2017;64(6):820-828
21. Malla N, Sengupta C, Dubey ML et al. Antigenae-mia and antibody response to Leishmania donovani stage-specific antigens and rk39 antigen in human immunodeficiency virus-infected patients. Br J Bi-omed Sci. 2003;60(4):210–6.
22. Hosseininejad M, Mohebali M, Hosseini F et al. Seroprevalence of canine visceral leishmaniasis in asymptomatic dogs in Iran. Iran J Vet Res. 2012;13(1):54–7.
23. Banu SS, Ahmed B-N, Shamsuzzaman AKM et al. Evaluation of recombinant K39 antigen and various promastigote antigens in sero-diagnosis of visceral leishmaniasis in Bangladesh. Parasite Epidemiol Control. 2016; 1(3):219–28.
24. Qu JQ, Zhong L, Masoom-Yasinzai M et al. Sero-diagnosis of Asian leishmaniasis with a recombinant antigen from the repetitive domain of a Leishmania kinesin. Trans R Soc Trop Med Hyg. 1994;88(5):543–5.
25. Ozensoy S, Ozbel Y, Turgay N et al. Serodiagnosis and epidemiology of visceral leishmaniasis in Tur-key. Am J Trop Med Hyg. 1998;59(3):363–9.
26. Rhalem A, Sahibi H, Guessous-Idrissi N et al. Im-mune response against Leishmania antigens in dogs naturally and experimentally infected with Leishmania infantum. Vet Parasitol. 1999;81(3):173–84.
27. Zerpa O, Ulrich M, Negrón E et al. Canine visceral leishmaniasis on Margarita Island (Nueva Esparta, Venezuela). Trans R Soc Trop Med Hyg. 2000;94(5):484–7.
28. Mettler M, Grimm F, Capelli G et al. Evaluation of Enzyme-Linked Immunosorbent Assays, an Im-munofluorescent-Antibody Test, and Two Rapid Tests (Immunochromatographic-Dipstick and Gel Tests) for Serological Diagnosis of Symptomatic and Asymptomatic Leishmania Infections in Dogs. J Clin Microbiol. 2005;43(11):5515–9
29. Quinnell RJ, Carson C, Reithinger R et al. Evalua-tion of rK39 Rapid Diagnostic Tests for Canine Visceral Leishmaniasis: Longitudinal Study and Me-ta-Analysis. PLoS Negl Trop Dis. 2013;7(1):e1992.
30. Rosário EY do, Genaro O, Franca-Silva JC et al. Evaluation of enzyme-linked immunosorbent assay using crude Leishmania and recombinant antigens as a diagnostic marker for canine visceral leishmaniasis. Mem Inst Oswaldo Cruz. 2005;100(2):197–203.
31. Porrozzi R, Santos da Costa MV, Teva A et al. Comparative Evaluation of Enzyme-Linked Im-munosorbent Assays Based on Crude and Recom-binant Leishmanial Antigens for Serodiagnosis of Symptomatic and Asymptomatic Leishmania infan-tum Visceral Infections in Dogs. Clin Vaccine Im-munol. 2007;14(5):544–8.
32. Edrissian GH, Ahanchin AR, Gharachahi AM et al. Seroepidemiological studies of visceral leishmaniasis and search for animal reservoirs in Fars province, southern Iran. Iran J Med Sci. 1993;18(3):99–105.
33. Badaró R, Reed SG, Barral A et al. Evaluation of the micro enzyme-linked immunosorbent assay (ELISA) for antibodies in American visceral leish-maniasis: antigen selection for detection of infec-tion-specific responses. Am J Trop Med Hyg. 1986;35(1):72–8.
34. Badaró R, Reed SG, Carvalho EM. Immunofluo-rescent antibody test in American visceral leishman-iasis: sensitivity and specificity of different morpho-logical forms of two Leishmania species. Am J Trop Med Hyg. 1983;32(3):480–4
35. Carvalho FA, Charest H, Tavares CAP et al. Diag-nosis of American visceral leishmaniasis in humans and dogs using the recombinant Leishmania do-novani A2 antigen. Diagn Microbiol Infect Dis. 2002;43(4):289–95.
36. Mohebali M, Malmasi A, Hajjaran H, et al. Dissem-inated Leishmaniasis Caused by Leishmania tropica in a Puppy from Karaj, Central Iran. Iran J Parasitol. 2011;6(2):69–73.
37. Hajjaran H, Mohebali M, Zarei Z et al. Leishmania tropica: Another Etiological Agent of Canine Visceral Leishmaniasis in Iran. Iran J Public Health. 2007;36(1):85–8.
Published
2020-03-09
How to Cite
1.
HOSSEINI FARASH BR, MOHEBALI M, KAZEMI B, HAJJARAN H, FATA A, RAOOFIAN R, AKHOUNDI B, MOJARRAD M, MASTROENI P, SHARIFI-YAZDI MK, TANIPOUR MH. The rK39 Antigen from an Iranian Strain of Leishmania infantum: Detection of Anti-Leishmania Antibodies in Humans and Dogs. Iran J Parasitol. 15(1):48-56.
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Original Article(s)