Development of a Multi-Epitope Recombinant Protein for the Diagnosis of Human Visceral Leishmaniasis
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Abstract
Background: Iran is one of the endemic areas of Mediterranean Visceral Leishmaniasis, a disease caused by Leishmania infantum. In this work, we examined whether Proteína quimérica 10 (PQ10) recombinant protein is suitable for immunological diagnosis of human visceral leishmaniasis.
Methods: The study was carried out in Tarbiat Modares University during 2016-2018. The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. Sequencing with proper primers was done, the expression, optimization of expression and protein purification were performed, and the purified recombinant protein was confirmed by western blot. The efficacy of PQ10 for serodiagnosis was evaluated with 50 positive and 50 negative serum samples, which confirmed by the direct agglutination test and collected from individuals living in the visceral leishmaniasis endemic areas of Iran. ELISA was performed with the PQ10 recombinant protein.
Results: The 95% CI sensitivity of ELISA that was evaluated with sera from naturally infected individuals was 84%. The 95% CI specificity value of the ELISA determined with sera from healthy individuals (50 serum samples) and from individuals with other infectious diseases was 82%. The 95% CI positive predictive value (PPV) and negative predictive value (NPV) were exterminated 82.35% and 83.67%, respectively.
Conclusion: We have used a recombinant synthetic protein to improve serodiagnosis of human visceral leishmaniasis. PQ10 could be useful for diagnosis of asymptomatic cases, as well as in the early phase of infections.
Cox FEG. History of human parasitology. Clinical Microbiology Reviews. 2002. p. 595–612.
WHO. Control of the leishmaniases. World Health Organization technical report series. Geneva: World Health Organization; 2010. xii p.
Kumar A. Leishmania and Leishmaniasis. New York, NY: Springer New York; 2013 [cited 2014 Jul 3]. 13-27 p. Available from: http://link.springer.com/10.1007/978-1-4614-8869-9
GhH E, Nadim A, Alborzi A V, Ardehali S. Visceral leishmaniasis: the Iranian experiences. Arch Iran Med. 1998;1(1):22–6.
Edrissian GH. Visceral leishmaniasis in Iran and the role of serological tests in diagnosis and epidemiological studies. Parasitol 21st Century ICOPA VIII, Izmir, Turkey CAB Int. 1996;63–78.
Mohebali M, Edrissian GHH, Nadim A, Hajjaran H, Akhoundi B, Hooshmand B, et al. Application of direct agglutination test (DAT) for the diagnosis and seroepide-miological studies of visceral leishmaniasis in Iran. Iran J Parasitol. 2006;1(1):15–25.
Mohebali M. Visceral leishmaniasis in Iran: Review of the Epidemiological and Clinical Features. Iran J Parasitol. Tehran University of Medical Sciences; 2013 Jul;8(3):348–58.
Mohebali M, Hajjaran H, Hamzavi Y, Mobedi I, Arshi S, Zarei Z, et al. Epidemiological aspects of canine visceral leishmaniosis in the Islamic Republic of Iran. Vet Parasitol. Elsevier; 2005;129(3):243–51.
Sundar S, More DK, Singh MK, Singh VP, Sharma S, Makharia A, et al. Failure of pentavalent antimony in visceral leishmaniasis in India: report from the center of the Indian epidemic. Clin Infect Dis. Oxford University Press; 2000;31(4):1104–7.
Motazedian M, Fakhar M, Motazedian MH, Hatam G, Mikaeili F. A urine-based polymerase chain reaction method for the diagnosis of visceral leishmaniasis in immunocompetent patients. 2008;60:151–4.
Sarkari B, Chance M, Hommel M. A capture ELISA for the diagnosis of visceral leishmaniasis using a monoclonal antibody against a leishmanial. Iran Biomed J. Iranian Biomedical Journal; 2005;9(3):117–22.
Sundar S, Rai M. Laboratory diagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol. Am Soc Microbiol; 2002;9(5):951–8.
Bittencourt D, Fraga M, Domingos E, Pacheco LV, Borja LS, Oliveira IQ De, et al. A multicentric evaluation of the recombinant Leishmania infantum antigen-based immunochromatographic assay for the serodiagnosis of canine visceral leishmaniasis. 2014;1–5.
Martins VT, Chávez-Fumagalli MA, Costa LE, Martins AMCC, Lage PS, Lage DP, et al. Antigenicity and protective efficacy of a Leishmania amastigote-specific protein, member of the super-oxygenase family, against visceral leishmaniasis. PLoS Negl Trop Dis. Public Library of Science; 2013;7(3):e2148.
Faria R, Giusta S, Jr GG, Penido MLO, Costa M, Gazzinelli RT. High-Throughput Analysis of Synthetic Peptides for the Immunodiagnosis of Canine Visceral Leishmaniasis. 2011;5(9).
Moura A, Rosa A, Tenório F, Rodrigues G, Alves R, Nagem P, et al. Acta Tropica Evaluation of three recombinant Leishmania infantum antigens in human and canine visceral leishmaniasis diagnosis. Acta Trop. Elsevier B.V.; 2014;137:25–30.
Akhoundi B, Mohebali M, Shojaee S, Jalali M, Kazemi B. Experimental Parasitology Rapid detection of human and canine visceral leishmaniasis : Assessment of a latex agglutination test based on the A2 antigen from amastigote forms of Leishmania infantum. Exp Parasitol. 2013;133(3):307–13.
Francisco R, Souza D, Luciolo Y, Souza R De, Borges-pereira L, Santana I, et al. Recombinant Leishmania ( Leishmania ) infantum Ecto-Nucleoside Triphosphate Diphosphohydrolase NTPDase-2 as a new antigen in canine visceral leishmaniasis diagnosis. Acta Trop [Internet]. Elsevier B.V.; 2013;125(1):60–6. Available from: http://dx.doi.org/10.1016/j.actatropica.2012.09.011
Faria R, Veloso LDC, Coura-vital W, Reis AB. Novel Recombinant Multiepitope Proteins for the Diagnosis of Asymptomatic Leishmania infantum- Infected Dogs. 2015;9(1):13–6.
Costa MM, Andrade HM, Bartholomeu DC, Freitas LM, Pires SF, Chapeaurouge AD, et al. Analysis of Leishmania chagasi by 2-D difference gel eletrophoresis (2-D DIGE) and immunoproteomic: identification of novel candidate antigens for diagnostic tests and vaccine. J Proteome Res. ACS Publications; 2011;10(5):2172–84.
Robinson CR, Sauer RT. Optimizing the stability of single-chain proteins by linker length and composition mutagenesis. Proc Natl Acad Sci. National Acad Sciences; 1998;95(11):5929–34.
el Harith A, Slappendel RJ, Reiter I, van Knapen F, De Korte P, Huigen E, et al. Application of a direct agglutination test for detection of specific anti-Leishmania antibodies in the canine reservoir. J Clin Microbiol. Am Soc Microbiol; 1989;27(10):2252–7.
WHO, editor. manual on visceral leishmaniasis control. first. World health organization; 1996. 84 p.
Tjitra E, Suprianto S, Dyer M, Currie BJ, Anstey NM. Field Evaluation of the ICT Malaria Pf/Pv Immunochromatographic Test for Detection of Plasmodium falciparum andPlasmodium vivax in Patients with a Presumptive Clinical Diagnosis of Malaria in Eastern Indonesia. J Clin Microbiol. Am Soc Microbiol; 1999;37(8):2412–7.
Antinori S, Calattini S, Longhi E, Bestetti G, Piolini R, Magni C, et al. Clinical use of polymerase chain reaction performed on peripheral blood and bone marrow samples for the diagnosis and monitoring of visceral leishmaniasis in HIV-infected and HIV-uninfected patients: a single-center, 8-year experience in Italy and review . Clin Infect Dis [Internet]. 2007 Jun 15 [cited 2014 Nov 7];44(12):1602–10. Available from: http://www.ncbi.nlm.nih.gov/pubmed/17516404
Srivastava P, Dayama A, Mehrotra S, Sundar S. Diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg. Royal Society of Tropical Medicine and Hygiene; 2011;105(1):1–6.
Mary C, Lamouroux D, Dunan S, Quilici M. Western blot analysis of antibodies to Leishmania infantum antigens: potential of the 14-kD and 16-kD antigens for diagnosis and epidemiologic purposes. Am J Trop Med Hyg. ASTMH; 1992;47(6):764–71.
Deborggraeve S, Laurent T, Espinosa D, Van der Auwera G, Mbuchi M, Wasunna M, et al. A simplified and standardized polymerase chain reaction format for the diagnosis of leishmaniasis. J Infect Dis. The University of Chicago Press; 2008;198(10):1565–72.
Elmahallawy EK, Martinez AS, Rodriguez-Granger J, Hoyos-Mallecot Y, Agil A, Mari JMN, et al. Diagnosis of leishmaniasis. J Infect Dev Ctries. 2014;8(08):961–72.
Seyyedtabaei SJ, Rostami A, Haghighi A, Mohebali M, Kazemi B, Fallahi S, et al. Detection of Potentially Diagnostic Leishmania Antigens with Western Blot Analysis of Sera from Patients with Cutaneous and Visceral Leishmaniases. Iran J Parasitol. 2017;12(2):206–14.
Costa CHN, Vieira JBF. Changes in the control program of visceral leishmaniasis in Brazil. Rev Soc Bras Med Trop. SciELO Brasil; 2001;34(2):223–8.
Sakkas H, Gartzonika C, Levidiotou S. Laboratory diagnosis of human visceral leishmaniasis. J Vector Borne Dis. National Institute of Malaria Research (ICMR); 2016;53(1):8.
Mohapatra TM, Singh DP, Sen MR, Bharti K, Sundar S. Compararative evaluation of rK9, rK26 and rK39 antigens in the serodiagnosis of Indian visceral leishmaniasis. J Infect Dev Ctries. 2009;4(02):114–7.
Soto M, Requena JM, Quijada L, Alonso C. Multicomponent chimeric antigen for serodiagnosis of canine visceral leishmaniasis. J Clin Microbiol. Am Soc Microbiol; 1998;36(1):58–63.
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. Clin Diagn Lab Immunol. Am Soc Microbiol; 2005;12(5):647–53.
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| Issue | Vol 16 No 1 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v16i1.5506 | |
| Keywords | ||
| Visceral leishmaniasis; Leishmania infantum PQ10 Protein Serodiagnosis Human Iran | ||
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