Abstract
Detection of Toxoplasma gondii in meat products, which is responsible for severe disease in humans and in animals, is a great challenge. Recombinant antibodies (rAbs) have been offered as valuable tools for different diagnostic and therapeutic goals. In this study, a latex agglutination assessment for identification of Toxoplasma gondii in contaminated meat products was developed. For this propose, recombinant anti-P30 single-chain Fv (scFv) antibody gene was cloned into pET26b vector to express this antibody in high level. This gene’s expression in Escherichia coli lead to production of significant amount of antibody (16 mg/L). After purification, the anti-P30 scFv was connected to beads of latex and used for agglutination test. These beads could agglutinate whole Toxoplasma gondii and contaminated meat samples. Thus, this rapid test may be applied to find contaminated samples.
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References
Anthony J, Near R, Sui-lam WIida E, Erns E, Wittekind M, Haber E, Ng SC (1992) Production of stable anti-digoxin Fv in Escherichia coli. Mol Immunol 29(10):1237–1247. https://doi.org/10.1016/0161-5890(92)90060-B
Aspinall TV, Marlee D, Hyde JE, Sims PF (2002) Prevalence of Toxoplasma gondii in commercial meat products as monitored by polymerase chain reaction—food for thought? Int J Parasitol 32(9):1193–1199. https://doi.org/10.1016/S0020-7519(02)00070-X
Ergİn S, Ciftcioglu G, Midilli K, Issa G, Gargİlİ A (2009) Detection of Toxoplasma gondii from meat and meat products by the nested-PCR method and its relationship with seroprevalence in slaughtered animals. Bull Vet Inst Pulawy 53(4):657–661
Golchin M, Khalili-Yazdi A, Karamouzian M, Abareghi A (2012) Latex agglutination test based on single-chain Fv recombinant antibody fragment. Scand J Immunol 75:38–45
Golchin M, Nakhaei-Moghadam M, Nourollahi-Fard SR (2015) Recombinant scFv antibodies against P30 surface protein of Toxoplasma Gondii. J. Kerman Univ Med Sci 22(6):602–613
Hill DE, Benedetto SM, Coss C, McCrary JL, Fournet VM, Dubey JP (2006) Effects of time and temperature on the viability of Toxoplasma gondii tissue cysts in enhanced pork loin. J Food Prot 69(8):1961–1965. https://doi.org/10.4315/0362-028X-69.8.1961
Hochuli E, Bannwarth W, Dobeli H, Gentz R, Stuber D (1988) Genetic approach to facilitate purification of recombinant proteins with a novel metal chelate adsorbent. Nat Biotech 6:1321–1325
Jones JL, Kruszon-Moran D, Sanders-Lewis K, Wilson M (2007) Toxoplasma gondii infection in the United States, 1999–2004, decline from the prior decade. Am J Trop Med Hyg 77(3):405–410
Kijlstra A, Jongert E (2009) Toxoplasma-safe meat: close to reality? Trends Parasitol 25(1):18–22. https://doi.org/10.1016/j.pt.2008.09.008
Kravetz JD, Federman DG (2005) Prevention of toxoplasmosis in pregnancy: knowledge of risk factors. Infect Dis Obstet Gynecol 13(3):161–165. https://doi.org/10.1080/10647440500068305
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685. https://doi.org/10.1038/227680a0
Opsteegh M, Langelaar M, Sprong H, den Hartog L, De Craeye S, Bokken G, Ajzenberg D, Kijlstra A, Van Der Giessen J (2010) Direct detection and genotyping of Toxoplasma gondii in meat samples using magnetic capture and PCR. Int J Food Microbiol 139:193–201
Pappas G, Roussos N, Falagas ME (2009) Toxoplasmosis snapshots: global status of Toxoplasma gondii seroprevalence and implications for pregnancy and congenital toxoplasmosis. Int J Parasitol 39:1385–1394
Saadatnia G, Golkar M (2012) A review on human toxoplasmosis. Scand J Infect Dis 44(11):805–814. https://doi.org/10.3109/00365548.2012.693197
Sambrook J, Russell DW (2001) Molecular cloning in a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Warnekulasuriya MR, Johnson JD, Holliman RE (1998) Detection of Toxoplasma gondii in cured meats. Int J Food Microbiol 45(3):211–215. https://doi.org/10.1016/S0168-1605(98)00158-5
Weisser NE, Hall JC (2009) Applications of single-chain variable fragment antibodies in therapeutics and diagnostics. Biotechnol Adv 27(4):502–520. https://doi.org/10.1016/j.biotechadv.2009.04.004
Yang J, Moyana T, MacKenzie S, Xia Q, Xiang J (1998) One hundred seventy-fold increase in excretion of an FV fragment-tumor necrosis factor alpha fusion protein (sFV/TNF-alpha) from Escherichia coli caused by the synergistic effects of glycine and triton X-100. AEM 64:2869–2874
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The authors would like to appreciate the members of Research as well as the staff of Microbiology Laboratory, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman.
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This research was financially supported by the Vice Chancellor for Research of Shahid Bahonar University of Kerman.
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Rezaei, A.A., Golchin, M. & Nakhaei Moghadam, M. Detection of Toxoplasma gondii in infected meat samples by a latex agglutination test using anti-P30 recombinant antibody. Comp Clin Pathol 28, 29–32 (2019). https://doi.org/10.1007/s00580-018-2638-0
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DOI: https://doi.org/10.1007/s00580-018-2638-0