Assessment of Recombinant A2-Latex Agglutination Test (RA2-LAT) and RA2-ELISA for Detection of Canine Visceral Leishmaniasis: A Comparative Field Study with Direct Agglutination Test in Northwestern Iran
-
Mahin FARAHMAND
,
Hossein NAHREVANIAN
,
Vahid KHALAJ
,
Mehdi MOHEBALI
,
Mohammad BARATI
,
Sanaz NADERI
,
Zabih ZAREI
,
Ghader KHALILI
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.
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.
| Files | ||
| Issue | Vol 13 No 2 (2018) | |
| Section | Original Article(s) | |
| Keywords | ||
| Recombinant A2 Latex agglutination test Canine visceral leishmaniasis ELISA Direct agglutination test Iran | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
