Abstract
Toxocara canis is an important gastrointestinal nematode of dogs and also a causative agent of visceral larva migrans in humans. Arginine kinase (AK) gene is one of the important biomolecule of phosphagen kinase of T. canis which is emerging as an exciting novel diagnostic target in toxocarosis. The present study was carried out to clone and characterize AK gene of T. canis for future utilization as a diagnostic molecule. Total RNA was extracted from intact adult worms and reverse transcription was done with oligo dT primers to obtain complementary DNA (cDNA). Polymerase chain reaction (PCR) was carried out using cDNA as template with specific primers which amplified a product of 1,202 bp. The amplicon was cloned into pDrive cloning vector and clone was confirmed by colony PCR and restriction endonuclease analysis. Sequence analysis of the gene showed 99.8 and 77.9 % homology with the published AK gene of T. canis (EF015466.1) and Ascaris suum respectively. Structural analysis shown that the mature AK protein consist of 400 amino acids with a molecular wt of 45360.73 Da. Further expression studies are required for producing the recombinant protein for its evaluation in the diagnosis of T. canis infection in humans as well as in adult dogs.
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References
Bachmeyer C, Lamarque G, Morariu R (2003) Visceral larva migrans mimicking lymphoma. Chest 123(4):1296–1297
Despommier D (2003) Toxocariasis: clinical aspects, epidemiology, medical ecology, and molecular aspects. Clin Microbiol Rev 16:265–272
Magnaval JF, Glickman LT, Dorchies P, Morassin B (2001) Highlights of human toxocariasis. Korean J Parasitol 39:1–11
Meyer RA, Sweeney HL, Kushmerick MJ (1984) A simple analysis of the phosphocreatine shuttle. Am J Physiol 246:C365–C377
Parkinson J, Mitreva M, Whitton C, Thomson M, Daub J, Martin J, Schmid R, Hall N (2004) A transcriptomic analysis of the phylum nematoda. Nat genet 36:1259–1267
Pawlowski Z (2001) Toxocariais in humans: clinical expression and treatment dilemma. J Helminthol 75:299–305
Sambrook J, Fritch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold spring harbor Laboratory press, New York
Wickramasinghe S, Uda K, Nagataki M, Yatawaraa L, Rajapakse RPVJ, Watanabe Y, Suzuki T, Agatsuma T (2007) Toxocara canis: molecular cloning, characterization, expression and comparison of the kinetics of cDNA-derived arginine kinase. Exp Parasitol 117:124–132
Wickramasinghe S, Yatawara L, Nagataki M, Takamoto M, Watanabe Y, Rajapakse RPVJ, Uda K, Suzuki T, Agatsuma T (2008) Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of T. canis for serodiagnosis of visceral larva migrans in the murine model. Parasitol Res 103:853–858
World Health Organization (WHO) (2004) Emerging zoonoses. www.who.in. Accessed 3–5 May 2004
Acknowledgments
The authors are thankful to the Director, Indian Veterinary Research Institute for providing all facilities to carry out the present investigation.
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Sahu, S., Samanta, S., Harish, D.R. et al. Molecular cloning and characterization of arginine kinase gene of Toxocara canis . J Parasit Dis 39, 211–215 (2015). https://doi.org/10.1007/s12639-013-0334-1
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DOI: https://doi.org/10.1007/s12639-013-0334-1