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Detection of Leishmania infantum Infection in Reservoir Dogs Using a Multiepitope Recombinant Protein (PQ10) | ||
| Archives of Razi Institute | ||
| مقاله 4، دوره 75، شماره 3، آذر 2020، صفحه 327-338 اصل مقاله (896.31 K) | ||
| نوع مقاله: Original Articles | ||
| شناسه دیجیتال (DOI): 10.22092/ari.2019.126524.1346 | ||
| نویسندگان | ||
| F. Jameie1؛ A. Dalimi* 1؛ M. Pirestani1؛ M. Mohebali2 | ||
| 1Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 2Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran | ||
| چکیده | ||
| Mediterranean visceral leishmaniasis is a zoonotic disease caused by Leishmania infantum and transmitted by the bites of infected female sand flies. Iran is one of the endemic areas of this disease. Dogs and canines are the major reservoir hosts of Leishmania infantum in the new and old world, including Iran. If visceral leishmaniasis is left untreated, it may result in a 90% mortality rate. The identification and elimination of infected dogs are efficient ways to control this disease. The diagnostic methods used to identify these animals cannot yield 100% detection. Therefore, in the present study, we used a multiepitope recombinant protein (PQ10) to distinguish between symptomatic and asymptomatic infections caused by Leishmania infantum in animal reservoirs (dogs). The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. In the following process, sequencing was performed with plasmid-specific primers and followed by the expression, optimization of expression. The purified recombinant protein was confirmed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. The efficacy of recombinant PQ10 protein was evaluated by an indirect enzyme-linked immunosorbent assay (ELISA) test using 150 dog sera (25 symptomatic positive sera, 25 asymptomatic positive sera, 80 negative sera, and 20 sera of dogs with other infectious diseases). Direct agglutination test (DAT) as the standard method was used to compare and determine specificity and sensitivity. The results indicated that the sensitivity and specificity of ELISA using infected dog sera were 94% and 86%, respectively. Positive and negative predictive values for dog sera were reported as 87.03% and 93.47%, respectively. This protein was able to identify 92% of asymptomatic dogs with visceral leishmaniasis. The results showed that the recombinant protein PQ10 is able to identify positive cases of canine visceral leishmaniasis, especially asymptomatic cases. | ||
| کلیدواژهها | ||
| Visceral leishmaniasis؛ Leishmania infantum؛ PQ10؛ Recombinant protein؛ Serodiagnosis؛ Dog؛ Iran | ||
| عنوان مقاله [English] | ||
| شناسایی عفونت لیشمانیا اینفانتوم در مخازن سگی با استفاده از یک پروتئین مولتی اپی توپ نوترکیب (PQ10) | ||
| چکیده [English] | ||
| لیشمانیوز احشایی مدیترانهای بیماری زئونوزی است که توسط لیشمانیا اینفانتوم ایجاد و با گزش پشه خاکی ماده آلوده انتقال مییابد. ایران یکی از مناطق اندمیک این بیماری میباشد. سگ و سگ سانان میزبانان مخزن اصلی لیشمانیا اینفانتوم در دنیای جدید و قدیم شامل ایران هستند. در صورتی که بیمار مبتلا به لیشمانیوز احشایی درمان نشود، در 90% موارد منجر به مرگ میشود. از راههای کنترل موثر این بیماری، شناسایی و حذف سگهای آلوده است. روشهای تشخیصی که برای شناسایی این حیوانات استفاده میشوند نمیتوانند 100% آنها را تشخیص دهند. به همین دلیل ما در این تحقیق جهت تشخیص عفونتهای علامتدار و بدون علامت ناشی از لیشمانیا اینفانتوم در مخازن حیوانی (سگ) از پروتئین نوترکیب مولتی اپی توپ PQ10 استفاده نمودیم. توالی کد کننده پروتئین نوترکیب PQ10 در وکتور بیانی pET28 ساب کلون شد و توسط شرکت GENERAY (چین) سنتز گردید. در مرحله بعد، توالی با پرایمرهای اختصاصی پلاسمید تعیین گردید و در نهایت مراحل بیان و بهینه سازی بیان صورت گرفت. پروتئین نوترکیب خالص با استفاده از SDS-PAGE و وسترن بلات مورد تایید قرار گرفت. ارزیابی کارآیی پروتئین نوترکیب PQ10 توسط آزمایش الایزای غیرمستقیم و با به کارگیری سرم 150 سگ (25 سرم مثبت از سگ علامت دار، 25 سرم مثبت از سگ بدون علامت، 80 سرم منفی و 20 سرم سگهای مبتلا به سایر بیماریهای عفونی) انجام شد. جهت مقایسه و تعیین اختصاصیت و حساسیت از آزمایش آگلوتیناسیون مستقیم (DAT) به عنوان روش استاندارد استفاده گردید. نتایج نشان داد که حساسیت و اختصاصیت الایزا با استفاده از سرم سگهای آلوده به ترتیب 94% و 86% است. ارزش اخباری مثبت و منفی در مورد سرم سگی به ترتیب 03/87% و 47/93% محاسبه گردید. این پروتئین قادر به شناسایی 92% از سگهای بدون علامت مبتلا به لیشمانیوز احشایی بود. نتایج نشان میدهد که پروتئین نوترکیب PQ10 قادر به شناسایی موارد مثبت لیشمانیوز احشایی سگی بالاخص موارد بدون علامت میباشد. | ||
| کلیدواژهها [English] | ||
| لیشمانیوز احشایی, لیشمانیا اینفانتوم, PQ10, پروتئین نوترکیب, تشخیص سرولوژیکی, سگ, ایران | ||
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| مراجع | ||
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Akhoundi, B., Mohebali, M., Shojaee, S., Jalali, M., Kazemi, B., Bandehpour, M., et al., 2013. 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 133, 307-313.
Badaró, R., Benson, D., Eulálio, M.C., Freire, M., Cunha, S., Netto, E.M., et al., 1996. rK39: A Cloned Antigen of Leishmania chagasi that Predicts Active Visceral Leishmaniasis. J Infect Dis 173, 758-761.
Costa, F.A.L., Goto, H., Saldanha, L.C.B., Silva, S.M.M.S., Sinhorini, I.L., Silva, T.C., et al., 2003. Histopathologic Patterns of Nephropathy in Naturally Acquired Canine Visceral Leishmaniasis. Vet Pathol 40, 677-684.
Costa, M.M., Andrade, H.M., Bartholomeu, D.C., Freitas, L.M., Pires, S.F., Chapeaurouge, A.D., et al., 2011. Analysis of Leishmania chagasi by 2-D difference gel electrophoresis (2-D DIGE) and immunoproteomic: identification of novel candidate antigens for diagnostic tests and vaccine. J Proteome Res 10, 2172-2184.
de Souza, R.F., Dos Santos, Y.L., de Souza Vasconcellos, R., Borges-Pereira, L., Caldas, I.S., de Almeida, M.R., et al., 2013. Recombinant Leishmania (Leishmania) infantum Ecto-Nucleoside Triphosphate Diphosphohydrolase NTPDase-2 as a new antigen in canine visceral leishmaniasis diagnosis. Acta Trop 125, 60-66.
Desjeux, P., 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27, 305-318.
Dye, C., Vidor, E., Dereure, J., 1993. Serological diagnosis of leishmaniasis: on detecting infection as well as disease. Epidemiol Infect 110, 647-656.
Edrissian, G., 1996. Visceral leishmaniasis in Iran and the role of serological tests in the diagnosis and epidemiological studies. J Kerman Univ Medical Sci 3, 97-108.
Edrissian, G., Nadim, A., Alborzi, A.V., Ardehali, S., 1998. Visceral leishmaniasis: the Iranian experiences. Arch Iran Med 1, 22-26.
Ehab Kotb, E., Antonio Sampedro, M., Javier, R.-G., Yannick, H.-M., Ahamd, A., Jose Mari Navarro, M., et al., 2014. Diagnosis of leishmaniasis. J Infect Dev Countr 8, 961-972.
el Harith, A., Slappendel, R.J., Reiter, I., van Knapen, F., de Korte, P., Huigen, E., et al., 1989. Application of a direct agglutination test for detection of specific anti-Leishmania antibodies in the canine reservoir. J Clin Microbiol 27, 2252-2257.
Evans, T.G., Vasconcelos, I.A.B., Lima, J.W., Teixeira, J.M., McAullife, I.T., Lopes, U.G., et al., 1990. Canine Visceral Leishmaniasis in Northeast Brazil: Assessment of Serodiagnostic Methods. Am J Trop Med Hyg 42, 118-123.
Farahmand, M., Khalaj, V., Mohebali, M., Khalili, G., Naderi, S., Ghaffarinejad, P., et al., 2015. Comparison of recombinant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in Northwestern Iran. Rev Soc Bras Med Trop 48, 188-193.
Faria, A.R., Costa, M.M., Giusta, M.S., Grimaldi, G., Jr., Penido, M.L., Gazzinelli, R.T., et al., 2011. High-throughput analysis of synthetic peptides for the immunodiagnosis of canine visceral leishmaniasis. PLoS Neglect Trop Dis 5, e1310.
Faria, A.R., de Castro Veloso, L., Coura-Vital, W., Reis, A.B., Damasceno, L.M., Gazzinelli, R.T., et al., 2015. Novel recombinant multiepitope proteins for the diagnosis of asymptomatic leishmania infantum-infected dogs. PLoS Neglect Trop Dis 9, e3429.
Fonseca, A.M., Faria, A.R., Rodrigues, F.T., Nagem, R.A., Magalhaes, R.D., Cunha, J.L., et al., 2014. Evaluation of three recombinant Leishmania infantum antigens in human and canine visceral leishmaniasis diagnosis. Acta Trop 137, 25-30.
Gharbi, M., Mhadhbi, M., Rejeb, A., Jaouadi, K., Rouatbi, M., Darghouth, M.A., 2015. Leishmaniosis (Leishmania infantum infection) in dogs. Rev Sci Tech 34, 613-626.
Gomez-Ochoa, P., Castillo, J.A., Lucientes, J., Gascon, M., Zarate, J.J., Arbea, J.I., et al., 2003. Modified direct agglutination test for simplified serologic diagnosis of leishmaniasis. Clin Diagn Lab Immunol 10, 967-968.
Gradoni, L., Maroli, M., Gramiccia, M., Mancianti, F., 1987 .Leishmania infantum infection rates in Phlebotomus perniciosus fed on naturally infected dogs under antimonial treatment. Med Vet Entomol 1, 339-342.
Lauricella, M.A., Maidana, C.G., Frias, V.F., Romagosa, C.M., Negri, V., Benedetti, R., et al., 2016. An rK28-Based Immunoenzymatic Assay for the Diagnosis of Canine Visceral Leishmaniasis in Latin America. Am J Trop Med Hyg 95, 92-98.
Mancianti, F., Falcone, M.L., Giannelli, C., Poli, A., 1995. Comparison between an enzyme-linked immunosorbent assay using a detergent-soluble Leishmania infantum antigen and indirect immunofluorescence for the diagnosis of canine leishmaniosis. Vet Parasitol 59, 13-21.
Mancianti, F., Pedonese, F., Poli, A., 1996. Evaluation of dot enzyme-linked immunosorbent assay (dot-ELISA) for the serodiagnosis of canine leishmaniosis as compared with indirect immunofluorescence assay. Vet Parasitol 65, 1-9.
Martínez Abad, L.P., Almeida, C.S., Mattos, A.M.M., Mendonça, A.C.P., Alves, M.J.M., Pinheiro, A.C., et al., 2017. Diagnostic accuracy of rKLO8 versus rK26 ELISAs for screening of canine visceral leishmaniasis. Acta Tropica 166, 133-138.
Martins, V.T., Chavez-Fumagalli, M.A., Costa, L.E., Canavaci, A.M., Martins, A.M., Lage, P.S., et al., 2013. Antigenicity and protective efficacy of a Leishmania amastigote-specific protein, member of the super-oxygenase family, against visceral leishmaniasis. PLoS Neglect Trop Dis 7, e2148.
Mary, C., Lamouroux, D., Dunan, S., Quilici, M., 1992. 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 47, 764-771.
Mettler, M., Grimm, F., Capelli, G., Camp, H., Deplazes, P., 2005. Evaluation of enzyme-linked immunosorbent assays, an immunofluorescent-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 43, 5515-5519.
Miro, G., Cardoso, L., Pennisi, M.G., Oliva, G., Baneth, G., 2008. Canine leishmaniosis--new concepts and insights on an expanding zoonosis: part two. Trends Parasitol 24, 371-377.
Mohebali, M., 2013. Visceral leishmaniasis in Iran: Review of the Epidemiological and Clinical Features. Iran J Parasitol 8, 348-358.
Mohebali, M., Edrisian, G., Nadim, A., Hajaran, H., Akhoundi, B., Houshmand, B., et al., 2006. Application of Direct Agglutination Test (DAT) for the diagnosis and sroepidemiological studies of visceral leishmaniasis in Iran. Iran J Parasitol 1, 15-25.
Molina, R., Amela, C., Nieto, J., San-Andrés, M., González, F., Castillo, J.A., et al., 1994. Infectivity of dogs naturally infected with Leishmania infantum to colonized Phlebotomus perniciosus. T Roy Soc Trop Med H 88, 491-493.
Nicolle, C., Comte, C., 1908. Origine canine du Kala-azar. Bull Soc Pathol Exot 1, 299-301.
Ordeix, L., Solano-Gallego, L., Fondevila, D., Ferrer, L., Fondati, A., 2005. Papular dermatitis due to Leishmania spp. infection in dogs with parasite-specific cellular immune responses. Vet Dermatol 16, 187-191.
Ozensoy, S., Ozbel, Y., Turgay, N., Alkan, M.Z., Gul, K., Gilman-Sachs, A., et al., 1998. Serodiagnosis and epidemiology of visceral leishmaniasis in Turkey. Am J Trop Med Hyg 59, 363-369.
Paranhos-Silva, M., Freitas, L.A., Santos, W.C., Grimaldi, G.J., Pontes-de-Carvalho, L.C., Oliveira-dos-Santos, A.J., 1996. A cross-sectional serodiagnostic survey of canine leishmaniasis due to Leishmania chagasi. Am J Trop Med Hyg 55, 39-44.
Pattabhi, S., Whittle, J., Mohamath, R., El-Safi, S., Moulton, G.G., Guderian, J.A., et al., 2010. Design, development and evaluation of rK28-based point-of-care tests for improving rapid diagnosis of visceral leishmaniasis. PLoS Negl Trop Dis 4.
Quinnell, R.J., Courtenay, O., Garcez, L., Dye, C., 1997. The epidemiology of canine leishmaniasis: transmission rates estimated from a cohort study in Amazonian Brazil. Parasitology 115, 143-156.
Rhalem, A., Sahibi, H., Guessous-Idrissi, N., Lasri, S., Natami, A., Riyad, M., et al., 1999. Immune response against Leishmania antigens in dogs naturally and experimentally infected with Leishmania infantum. Vet Parasitol 81, 173-184.
Ribeiro, R.R., Michalick, M.S.M., da Silva, M.E., Dos Santos, C.C.P., Frezard, F.J.G., da Silva, S.M., 2018. Canine Leishmaniasis: An Overview of the Current Status and Strategies for Control. BioMed Res Int 2018, 1-12.
Robinson, C.R., Sauer, R.T., 1998. Optimizing the stability of single-chain proteins by linker length and composition mutagenesis. Proc Natl Acad Sci U S A 95, 5929-5934.
Rodriguez-Cortes, A., Ojeda, A., Francino, O., Lopez-Fuertes, L., Timon, M., Alberola, J., 2010. Leishmania infection: laboratory diagnosing in the absence of a "gold standard". Am J Trop Med Hyg 82, 251-256.
Roffi, J., Dedet, J.P., Desjeux, P., Garre, M.T., 1980. Detection of circulating antibodies in cutaneous leishmaniasis by enzyme-linked immunosorbent assay (ELISA). Am J Trop Med Hyg 29, 183-189.
Sakkas, H., Gartzonika, C., Levidiotou, S., 2016. Laboratory diagnosis of human visceral leishmaniasis. J Vector Borne Dis 53, 8-16.
Santarem, N., Silvestre, R., Cardoso, L., Schallig, H., Reed, S.G., Cordeiro-da-Silva, A., 2010. Application of an improved enzyme-linked immunosorbent assay method for serological diagnosis of canine leishmaniasis. J Clin Microbiol 48, 1866-1874.
Scalone, A., De Luna, R., Oliva, G., Baldi, L., Satta, G., Vesco, G., et al., 2002. Evaluation of the Leishmania recombinant K39 antigen as a diagnostic marker for canine leishmaniasis and validation of a standardized enzyme-linked immunosorbent assay. Vet Parasitol 104, 275-285.
Seyyedtabaei, S.J., Rostami, A., Haghighi, A., Mohebali, M., Kazemi, B., Fallahi, S., et al., 2017. Detection of Potentially Diagnostic Leishmania Antigens with Western Blot Analysis of Sera from Patients with Cutaneous and Visceral Leishmaniases. Iran J Parasitol 12, 206-214.
Srivastava, P., Dayama, A., Mehrotra, S., Sundar, S., 2011. Diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg 105, 1-6.
Sundar, S., Rai, M., 2002. Laboratory diagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol 9, 951-958.
Tjitra, E., Suprianto, S., Dyer, M., Currie, B.J., Anstey, N.M., 1999. Field evaluation of the ICT malaria P.f/P.v immunochromatographic test for detection of Plasmodium falciparum and Plasmodium vivax in patients with a presumptive clinical diagnosis of malaria in eastern Indonesia. J Clin Microbiol 37, 2412-2417.
Travi, B.L., Cordeiro-da-Silva, A., Dantas-Torres, F., Miro, G., 2018. Canine visceral leishmaniasis: Diagnosis and management of the reservoir living among us. PLoS Neglect Trop Dis 12, e0006082.
Venturin, G.L., Bragato, J.P., Silva, K.L., de Lima, V.M., 2015. Recombinant K28 antigen in ELISA in the diagnosis of canine visceral leishmaniosis. Parasite Immunol 37, 670-673.
WHO, 1996. Manual on visceral leishmaniasis control. World Health Organization, Geneva. | ||
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