The Use of Recombinant 31 kDa Antigens of Trichinella spiralis for Serodiagnosis of Experimental Trichinellosis
-
Li Wang
,
Xiang Yu Tian
,
Ge Ge Sun
,
Ruo Dan Liu
,
Li Na Liu
,
Xi Zhang
,
Peng Jiang
,
Zhong Quan Wang
,
Jing Cui
Abstract
Background:We have previously reported that a 31 kDa protein was screened from the excretory-secretory (ES) proteins of Tichinella spiralis muscle larvae (ML) by immunoproteomics using early infection sera, and the gene encoding a 31 kDa protein from T. spiralis was cloned and expressed in an E. coli expression system. In this study, the recombinant 31 kDa antigens were used for detection of anti- Trichinella antibodies in serum of experimentally infected mice by ELISA.
Methods:Anti-Trichinella IgG antibodies in sera of mice infected with Trichinella were assayed by ELISA with recombinant 31 kDa antigens, and its sensitivity and specificity were compared with ELISA with ES antigen.
Results:The sensitivity and specificity of ELISA with recombinant antigens was 96.67% (29/30) and 96.87% (62/64), compared with 100% (30/30) and 98.44% (63/64) of ELISA with ES antigens was (P＞0.05). In heavily, moderately and lightly infected mice (500, 300 and 100 larvae/mouse), anti-Trichinella antibodies were firstly detected by ELISA with recombinant antigens at 8, 12 and 14 dpi, re-spectively; then increased rapidly with a detection rate of 100% respectively at 28, 22 and 30 dpi. While the antibodies were firstly detected by ELISA with ES anti-gens at 10, 8 and 10 dpi, respectively, the antibody positive rate reached 100% at 14, 12 and 22 dpi, respectively.
Conclusion:The recombinant 31 kDa antigens of T. spirali had a good sensitivity and specificity for detecting anti-Trichinella antibodies and might be the potential diagnostic antigen for trichinellosis.
Murrell KD, Pozio E. Worldwide occurrence and impact of human trichinellosis, 1986-2009. Emerg Infect Dis. 2011; 17:2194-2202.
Iran J Parasitol: Vol. 10, No. 2, Apr -Jun 2015, pp.230-237
Available at: http://ijpa.tums.ac.ir
Cui J, Jiang P, Liu LN, Wang ZQ. Survey of Trichinella infections in domestic pigs from northern and eastern Henan, China. Vet Parasi-tol. 2013; 194:133-135.
Cui J, Wang ZQ, Kennedy MW. The re-emer-gence of trichinellosis in China? Trends Parasi-tol. 2006; 22:54-55.
Dupouy-Camet J. Presidential address of ICT12 Conference: “Trichinella and trichinello-sis-a never ending story”. Vet Parasitol. 2009; 159:194-196.
Cui J, Wang ZQ, Xu BL: The epidemiology of human trichinellosis in China during 2004-2009. Acta Trop. 2011; 118:1-5.
Gomez-Morales MA, Ludovisi A, Amati M, Cherchi S, Pezzotti P, Pozio E: Validation of an enzyme-linked immunosorbent assay for di-agnosis of human trichinellosis. Clin Vaccine Immunol. 2008; 15:1723-1729.
Gottstein B, Pozio E, Nockler K: Epidemiol-ogy, diagnosis, treatment, and control of trichi-nellosis. Clin Microbiol Rev. 2009; 22:127-145.
Mahannop P, Setasuban P, Morakote N, Tap-chaisri P, Chaicumpa W. Immunodiagnosis of human trichinellosis and identification of spe-cific antigen for Trichinella spiralis. Int J Parasi-tol.1995; 25:87-94.
Yera H, Andiva S, Perret C, Limonne D, Boireau P, Dupouy-Camet J: Development and evaluation of a Western blot kit for diagno-sis of human trichinellosis. Clin Diagn Lab Immunol. 2003; 10: 793-796.
Kapel CM and Gamble HR. Infectivity, persis-tence, and antibody response to domestic and sylvatic Trichinella spp. in experimentally in-fected pigs. Int J Parasitol. 2000; 30:215–221.
Gamble HR, Pozio E, Bruschi F, Nockler K, Kapel CM, Gajadhar AA. International Com-mission on Trichinellosis: recommendations on the use of serological tests for the detection of Trichinella infection in animals and man. Parasite. 2004; 11:3-13.
Liu LN, Jing FJ, Cui J, Fu GY, Wang ZQ. De-tection of circulating antigen in serum of mice infected with Trichinella spiralis by an IgY-IgM mAb sandwich ELISA. Exp Parasitol. 2013; 133:150-155.
Bruschi F, Moretti A, Wassom D, and Piergili-Fioretti D. The use of a synthetic antigen for the serological diagnosis of human trichinello-sis. Parasite. 2001; 8:S141-S143.
Dea-Ayuela MA, Romarís F, Ubeira FM, Ra-ma-Iñiguez S, Martínez-Fernández AR, Bolás F. Possible presence of common tyvelose-containing glycans in Trichinella L1 larvae and embryonated eggs of several nematodes. Para-site. 2001; 8:S120-S122.
Zarlenga DS, Gamble HR. Molecular cloning and expression of an immunodominant 53-kDa excretory–secretory antigen from Trichi-nella spiralis muscle larvae. Mol Biochem Parasi-tol. 1995; 72:253.
Jung D, Teifke JP, Karger A, Michael K, Venz S, Wittmann W, Kindermann K, Nöckler K, Mundt E. Evaluation of baculovirus-derived recombinant 53-kDa protein of Trichinella spi-ralis for detection of Trichinella-specific anti-bodies in domestic pigs by ELISA. Parasitol Res. 2007; 100: 429-437.
Su XZ, Prestwood AK, McGraw RA. Cloning and expression of complementary DNA en-coding an antigen of Trichinella spiralis. Mol Bio-chem Parasitol. 1991; 45:331–336.
Wang B, Wang ZQ, Jin J, Ren HJ, Liu LN, Cui J. Cloning, expression and characterization of a Trichinella spiralis serine protease gene encoding a 35.5 kDa protein. Exp Parasitol. 2013;134:148-154.
Wang R, Wang ZQ, Cui J. Immunodiagnostic value and immune protection of the recombi-nant Ts21 antigen of Trichinella spiralis. Chin J Parasitol Parasit Dis. 2009; 27: 17-21.
Arasu P, Ellis LA, Iglesias R, Ubeira FM, Appleton JA. Molecular analysis of antigens targeted by protective antibodies in rapid expulsion of Trichinella spiralis. Mol Biochem Parasitol. 1994; 65: 201-211.
Wang L, Wang ZQ, Hu DD, Cui J. Proteomic analysis of Trichinella spiralis muscle larval excre-tory-secretory proteins recognized by early in-fection sera. Biomed Res Int. 2013; 2013:139745.
Wang L, Cui J, Hu DD, Liu RD, Wang ZQ. Identification of early diagnostic antigens from major excretory-secretory proteins of Trichinella spiralis muscle larvae using immunoproteomics. Parasit Vectors. 2014; 7:40.
Cui J, Wang L, Sun GG, Liu LN, Zhang SB, Liu RD, Zhang Xi, Jiang P, Wang ZQ. Charac-terization of a Trichinella spiralis 31 kDa protein and its potential application for the serodi-
Wang et al.: The Use of Recombinant 31 kDa Antigens of Trichinella spiralis …
Available at: http://ijpa.tums.ac.ir 237
agnosis of trichinellosis. Acta Trop. 2015; 142:57-63.
Wang ZQ, Fu GY, Jing FJ, Jin J, Ren HJ, Jiang P, Cui J. Detection of Trichinella spiralis circulat-ing antigens in serum of experimentally in-fected mice by an IgY-mAb sandwich ELISA. Foodborne Pathog Dis. 2012; 9:727-933.
Gamble HR, Bessonov AS, Cuperlovic K, Gajadhar AA, van Knapen F, Noeckler K, Schenone H, Zhu X. International Commis-sion on Trichinellosis: recommendations on methods for the control of Trichinella in domes-tic and wild animals intended for human con-sumption. Vet Parasitol. 2000; 93:393-408.
Li F, Cui J, Wang ZQ, Jiang P. Sensitivity and optimization of artificial digestion in the in-spection of meat for Trichinella spiralis. Food-borne Pathog Dis. 2010; 7:879-885.
Wang SW, Wang ZQ, Cui J. Protein change of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae. Parasitol Res.2011; 108:593-599.
Hu DD, Cui J, Wang L, Liu LN, Wei T, Wang ZQ. Immunoproteomic analysis of the excre-tory-secretory proteins from Spirometra mansoni sparganum. Iran J Parasitol. 2013; 8:408-416.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72: 248-254.
Cui J, Li N, Wang ZQ, Jiang P, Lin XM. Sero-diagnosis of experimental sparganum infec-tions of mice and human sparganosis by ELI-
SA using ES antigens of Spirometra mansoni spargana. Parasitol Res. 2011; 108: 1551-1556.
Wang ZQ, Lin XM, Zhang HW, Xu BL, Zhang X, Jiang P, Cui J. Serological survey for sparganum infection in people of central China. Helminthiologia. 2014: 51:158-161.
Zhang YW, Lee DL, Smith JE. Biochemical characterisation of Trichinella spiralis and T. pseu-dospiralis stichocyte antigens. Appl Parasi-tol.1993; 34:291-294.
Kapel CM, Gamble HR. Infectivity, persistence, and antibody response to domestic and sylvatic Trichinella spp. in experimentally infected pigs. Int J Parasitol. 2000; 30: 215-221.
Appleton JA, Bell RG, HOMAN W, Van Knapen F. Consensus on Trichinella spiralis anti-gens and antibodies. Parasitol Today. 1991; 7: 190-192.
Nagano I, Wu Z, Takahashi Y. Species-specific antibody responses to the recombinant 53-kilo-dalton excretory and secretory proteins in mice infected with Trichinella spp. Clin Vaccine Im-munol. 2008; 15:468-473.
Yang Y, Qin W, Qiu H, Liu Y. Characteriza-tion of TsDAF-21/HSP90 protein from the parasitic nematode Trichinella spiralis. Parasitol Res. 2014;113:2209-2217.
Salinas-Tobon Mdel R, Navarrete-Leon A, Mendez-Loredo BE, Esquivel-Aguirre D, Mar-tínez-Abrajan DM, Hernandez-Sanchez J. Trichinella spiralis: strong antibody response to a 49 kDa newborn larva antigen in infected rats. Exp Parasitol. 2007; 115: 160-167.
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| Issue | Vol 10 No 2 (2015) | |
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| Serodiagnosis Trichinella spiralis Trichinellosis | ||
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