Journal of Veterinary Parasitology, 27(2) 2013 : 100-104 Molecular characterization of Ag2 gene of Taenia solium Bareilly isolate H.V. Manjunathachar, B.C. Saravanan1, A.K. Tewari, M. Sankar, O.K. Raina, S.C. Gupta, B.L. Balaraju and M. Kesavan Division of Parasitology, Indian VeterinaryResearchInstitute, Izatnagar, 243122, India Abstract Taenia solium cysticerci infection is common in pigs and human beings and is a major health problem in many developing countries of Latin America, Africa and Asia. In the present communication we report the molecular cloning and analysis of the 220 bp coding sequence for Ag2 protein of T. solium cysticerci (Bareilly isolate). Using cDNA template the entire coding sequence (devoid of signal sequence) for Ag2, was PCR amplified and ligated to pTZ57R/T cloning vector. The recombinant plasmid was extracted from the white colonies and purified for custom sequencing for nucleotides. The nucleotide sequence information revealed a relatively small molecular size of Ag2 protein composed of 67 amino acids with theoretical molecular weight and isoelectric point of 7.67 kDa and 9.42, respectively. The nucleotide and deduced amino acid sequences of the Bareilly isolate was aligned and analyzed in silico using DNA STAR and MEGA version 4.0 softwares with the related sequences from other isolates of T. solium available in public domain. The nucleotide sequence of T. solium cysticerci Ag2 (Bareilly isolate) revealed 99.5 to 100% sequence homology with the publisbed sequences of T. solium cysticerci China, Brazil, Cameroon, Tanzania and South Africa isolates. The near hundred per cent sequence homology of Ag2 gene from different isolates of T. solium, viz. China, Brazil, Cameroon, Tanzania, South Africa and India, proved its highly conserved nature and therefore we predict its potential application in sensitive serodiagnosis of taeniasis. Keywords: Taenia solium, Ag2 gene, Cloning, Cysticercus cellulosae, Pig. Introduction Cysticercosis in pigs is caused by Cysticercus cellulosae, the metacestode of Taenia solium. The infection is responsible for neurocysticercosis(NCe) in human beings, an important zoonotic disease. Taenia solium cysticercosis in pigs and humans is endemic in many developing countries of Latin America, Africa and Asia (Sarti et al., 1992; Flisser, 2002; Geerts et aZ., 2002; Ito et aI., 2002) and is responsible for huge economic loss to the pig industry due to carcass condemnation(Acevedo, 1982;Flisser et at., 1986;Pawlowski et aZ., 2005; Flisser et aZ., 2006; Deckers and Domy, 2010). Cysticercosis and neurocysticercosis in humans has been documented from various parts of India reflecting its high prevalence in the susceptible population. Diagnosis of porcine cysticercosis is important for identificationof endemic regions (Gonzalez et al., 1994, 1999 'Corresponding author E-mail: drbcsaravanan@gmail.com 2Division of Temperate Animal Husbandry, Mukteswsar J. Vet. Parasitol. 200-0064-480/$2.50 lid lAA VP, India and 2001). Further, an unequivocal detection of cysticercosis in humans is also critical for specific chemotherapeutic intervention (Hancock et aZ., 2006). Development of a specific and sensitive diagnostic test for pigs would be a major advancement over the conventional relatively insensitive method of carcass/meat inspection based detection system for cysticercosis. Establishing the sero-prevalence of cysticercosis through the detection of anti-parasite antibodies using a high throughput serodiagnostic test will be a valuable epidemiological tool particularly for large scale application in endemic areas (Harrison et aZ., 1989; Lightowlers, 1990). For a rapid and efficient diagnosis of pig cysticercosis, cyst fluid derived purified glycoproteins are promising molecules however, ensuring the supply of the native glycoproteins in abundant quantities is a major constraint (Cai et aZ., 2006; Deckers and Domy, 2010). Therefore, development of an efficient, reproducible and economic diagnostic test that precludes the use of crude parasite material is essential (Chung et at. 1999; Greene et at. 2000; Hancock et at. 2003; 100