Assessment of Recombinant A2-Latex Agglutination Test (RA2-LAT) and RA2-ELISA for Detection of Canine Visceral Leishman-iasis: A Comparative Field Study with Direct Agglutination Test in Northwestern Iran

  • Mahin FARAHMAND Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran
  • Hossein NAHREVANIAN Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran
  • Vahid KHALAJ Dept. of Biotechnology, Pasteur Institute of Iran, Tehran, Iran
  • Mehdi MOHEBALI Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad BARATI Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Infectious Diseases Research Center, AJA University of Medical Sciences, Tehran, Iran
  • Sanaz NADERI Islamic Azad University, Science and Research Branch of Kurdistan, Sanandaj, Iran
  • Zabih ZAREI Meshkin-Shahr Research Station, School of Public Health, Tehran University of Medical Sciences, Meshkin-Shahr, Iran
  • Ghader KHALILI Dept. of Immunology, Pasteur Institute of Iran, Tehran, Iran
Keywords: Recombinant A2, Latex agglutination test, Canine visceral leishmaniasis, ELISA, Direct agglutination test, Iran

Abstract

Background: This study aimed to set-up latex agglutination test (LAT) and ELISA based on recombinant A2 from Iranian strain of Leishmania (L.) infantum (rA2-Ag) and evaluated for detection of anti- Leishmania antibodies in dogs compared to standard direct agglutination test (DAT).Methods: The rA2-Ag was synthesized under a part of the A2 gene sequences which contain immune dominant sequences and less number of repetitive sequences. Latex beads, 0.8 μm (Sigma, USA) were sensitized with rA2-Ag. The tests were carried out on sera collected from 350 ownership dogs including symptomatic (n=67), asymptomatic (n=230) canine visceral leishmaniasis (CVL), and (n=53) uninfected domestic dogs as control group.Results: Anti-leishmanial antibodies were detected in 97 (27.7%), 96 (27.4%) and 29 (%9) of the serum samples by using DAT, rA2-ELISA, and rA2-latex, respectively with ≥1:320 as a cut-off titer when DAT-confirmed cases were compared with the control groups. A combined sensitivity of 52% and specificity of 82.40% for rA2-ELISA and 23.8% and specificity 95.38%, respectively were found with ≥1:320 as a cut-off titer when DAT-confirmed cases were compared with the control groups. The concordance between rA2-ELISA and rA2 latex compared with DAT as a gold standard serological test for VL were found 73.7% and 77.5%, respectively.Conclusion: A good degree of agreement was found between rA2-ELISA and DAT (73.7%). rA2-ELISA could detect more seropositive serum samples than rA2-LAT and it may be recommended as an alternative tool for the diagnosis of CVL.

References

WHO. Leishmaniasis in high-burden countries: an epidemiological update based on data reported in 2014. Week Epidemi-olog Rec. 2016;91(22):287-96.

Edrissian Gh H, Nadim A, Alborzi AV, Ardehali S. Visceral leishmaniasis: the Ira-nian experiences. Arch Iranian Med. 1998; 1: 22-6.

Mohebali M. Visceral leishmaniasis in Iran: Review of the Epidemiological and Clinical Feature. Iran J Parasitol: 2013; 8(3):348-58.

Moshfe A, Mohebali M, Edrissian GhH et al. Canine visceral leishmaniasis: Asymp-tomatic infected dogs as a source of L. infantum infection. Acta Trop. 2009;112:101–5.

Farahmand M, Nahrevanian H. Applica-tion of Recombinant Proteins for Serodi-agnosis of Visceral Leishmaniasis in Hu-mans and Dogs. Iran Biomed J. 2016; 20(3): 128-34.

Mancianti F, Meciani N. Specific serodiag-nosis of canine leishmaniasis by indirect immunofluorescence, indirect hemaggluti-nation, and counter-immuno-electrophoresis. Am J Vet Res. 1988; 49(8): 1409-11.

Harith A, Kolk AH, Kager PA, Leeuwen-burg J, Muiga R, Kiugu S, Laarman JJ. A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leish-maniasis. Trans R Soc Trop Med Hyg. 1986; 80(4): 583-6.

Srivastava P, Dayama A, Mehrotra S, Sundar SH. Diagnosis of visceral leish-maniasis. Trans R Soc Trop Med Hyg. 2011; 105(1): 1–6.

Burns JM, Schreffler WG, Benson DR, Ghalib WH, Badaro´ R, Reed SG. Molecu-lar characterization of a kinesin- related an-tigen of Leishmania chagasi that detects spe-cific antibodies in African and American visceral leishmaniasis. Proc Natl Acad Sci. USA. 1993; 90(2): 775-9.

Mohebali M, Taran M, Zareii Z. Rapid detection of Leishmania infantum infection in dogs: Comparative study using immuno-chromatographic dipstickrK39 test and di-rect agglutination. Vet Parasitol. 2004;121(3,4): 239-45.

Farahmand M, Khalaj V, Mohebali M, Khalili G, Naderi S, Ghaffarinejad P, Nahrevanian H. Comparison of recombi-nant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in North-western Iran. Rev Soc Bras Med Trop. 2015; 48(2): 188-93.

Babakhan L, Mohebali M, Akhoundi B, Edrissian GhH, Keshavarz H. Rapid de-tection of Leishmania infantum infection in dogs: a comparative study using fast agglu-tination screening test (FAST) and direct agglutination test (DAT) in Iran. Parasitol Res. 2009; 105:717- 20.

Mohebali M, Edrissian GhH, Nadim A, Hajjaran H, Akhoundi B, Hooshmand B. Application of direct agglutination test (DAT) for the diagnosis and seroepidemi-ological studies of visceral leishmaniasis in Iran. Iran J Parasitol. 2006; 1:15-25

Molina-Boliva JA, Galisteo-Gonzalez RF. Latex immunoagglutination assays. J Mac-romol Sci Pare-C: Polymer Rev.2005; 45:59-98.

Laemmli, UK. Cleavage of structural pro-teins during the assembly of the head of bacteriophage T4. Nature. 1970; 227: 680–5.

Babaie J, Miri M, Sadeghiani GH, Zare M, Khalili GH, Golkar M. Expression and Single-step Purification of GRA8 Antigen of Toxoplasma gondii in Escherichia coli. Avi-cenna J Med Biotech. 2011; 3(2): 67-77.

Akhoundi B, Mohebali M, Shojaee S et al. Rapid detection of human and canine vis-ceral leishmaniasis: Assessment of a latex agglutination test based on the A2 antigen from amastigote forms of Leishmania infantum. Exp Parasitol. 2013; 133: 307–13.

Mohebali M, Edrissian GhH, Shirzadi MR et al. An observational study for current distribution of visceral leishmaniasis in dif-ferent geographical zones of Iran for implication of health policy. Travel Med Infect Dis. 2011; 9:67-74.

Mohebali M, Hajjaran H, Hamzavi Y et al. Epidemiological aspects of canine visceral leishmaniosis in the Islamic Republic of Iran. Vet Parasitol. 2005; 129: 243-51.

Bradford MM. Rapid and sensitive meth-od for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Bio-chem. 1976; 72: 248- 54.

Reithinger R, Quinnell RJ, Alexander BC, Davies R. Rapid detection of Leishmania in-fantum infection in dogs: comparative study using an immunochromatographic dip-stick test, enzyme-linked immunosorbent assay, and PCR. J Clin Microbiol. 2002; 40: 2352-6.

Porrozzi R, Santos da Costa MV, Teva A et al. Comparative evaluation of enzyme-linked immunosorbent assays based on crude and recombinant leishmanial anti-gens for serodiagnosis of symptomatic and asymptomatic Leishmania infantum vis-ceral infections in dogs. Clin Vaccine Im-munol. 2007;14(5):544-8.

Akhoundi B. Preparation of agglutination antigen based on dominant antigenic component of axenic amastigote and promastigote of Iranian strain of L. infan-tum and their evaluation for diagnosis of VL in endemic areas of Iran. Ph.D. thesis. 2011.

Farahmand M, Atashi Shirazi H, Nahrevanian H, Hajjaran H. Molecular analysis of A2-genes encoding stage-specific S antigen-like proteins among iso-lates from Iranian cutaneous and visceral leishmaniasis. Iran J Basic Med Sci, 2011; 14(5):407-13.

Farahmand M, Nahrevanian H, Assmar M, Mohebali M, Zarei Z. Expression of A2 proteins in amastigotes of Leishmania infantum produced from canine isolates col-lected in the district of Meshkinshahr, in north–western Iran. Ann Trop Med Para-sitol, 2008; 102(1): 81-4.

Published
2018-06-25
How to Cite
1.
FARAHMAND M, NAHREVANIAN H, KHALAJ V, MOHEBALI M, BARATI M, NADERI S, ZAREI Z, KHALILI G. Assessment of Recombinant A2-Latex Agglutination Test (RA2-LAT) and RA2-ELISA for Detection of Canine Visceral Leishman-iasis: A Comparative Field Study with Direct Agglutination Test in Northwestern Iran. IJPA. 13(2):172-9.
Section
Original Article(s)