title: A proline racemase based PCR for identification of Trypanosoma vivax in cattle blood.
abstract: A study was conducted to develop a Trypanosoma vivax (T. vivax) specific PCR based on the T. vivax proline racemase (TvPRAC) gene. Forward and reverse primers were designed that bind at 764-783 bp and 983-1002 bp of the gene. To assess its specificity, TvPRAC PCR was conducted on DNA extracted from different haemotropic pathogens: T. vivax from Nigeria, Ethiopia and Venezuela, T. congolense Savannah type, T. brucei brucei, T. evansi, T. equiperdum, T. theileri, Theileria parva, Anaplasma marginale, Babesia bovis and Babesia bigemina and from bovine, goat, mouse, camel and human blood. The analytical sensitivity of the TvPRAC PCR was compared with that of the ITS-1 PCR and the 18S PCR-RFLP on a dilution series of T. vivax DNA in water. The diagnostic performance of the three PCRs was compared on 411 Ethiopian bovine blood specimens collected in a former study. TvPRAC PCR proved to be fully specific for T. vivax, irrespective of its geographical origin. Its analytical sensitivity was lower than that of ITS-1 PCR. On these bovine specimens, TvPRAC PCR detected 8.3% T. vivax infections while ITS-1 PCR and 18S PCR-RFLP detected respectively 22.6 and 6.1% T. vivax infections. The study demonstrates that a proline racemase based PCR could be used, preferably in combination with ITS-1 PCR, as a species-specific diagnostic test for T. vivax infections worldwide.
authors.affiliation: College of Veterinary Medicine and Agriculture, Addis Ababa University, Debre Zeit, Ethiopia ; Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium ; Department Biosystems, Faculty of Bioscience Engineering, Katholieke Universiteit Leuven, Leuven, Belgium.
authors.forename: Regassa
authors.initials: R
authors.lastname: Fikru
authors.affiliation: College of Veterinary Medicine and Agriculture, Addis Ababa University, Debre Zeit, Ethiopia.
authors.forename: Ashenafi
authors.initials: A
authors.lastname: Hagos
authors.affiliation: Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors.forename: Stijn
authors.initials: S
authors.lastname: Rogé
authors.affiliation: Grupo de Inmunobiología, Centro de Estudios Biomédicos y Veterinarios, Universidad acional Experimental Simón Rodríguez, Caracas, Venezuela.
authors.forename: Armando
authors.initials: A
authors.lastname: Reyna-Bello
authors.affiliation: Grupo de Bioquímica e Inmunología de Hemoparásitos, Departamento de Biología Celular, Universidad Simón Bolívar, Caracas, Venezuela.
authors.forename: Mary Isabel
authors.initials: MI
authors.lastname: Gonzatti
authors.affiliation: College of Veterinary Medicine and Agriculture, Addis Ababa University, Debre Zeit, Ethiopia.
authors.forename: Bekana
authors.initials: B
authors.lastname: Merga
authors.affiliation: Department Biosystems, Faculty of Bioscience Engineering, Katholieke Universiteit Leuven, Leuven, Belgium.
authors.forename: Bruno Maria
authors.initials: BM
authors.lastname: Goddeeris
authors.affiliation: Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors.forename: Philippe
authors.initials: P
authors.lastname: Büscher
mesh_terms: D000591:Amino Acid Isomerases / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000097:blood* / Q000469:parasitology*; D002418:Cattle Diseases / Q000097:blood / Q000469:parasitology; D006801:Humans; D057230:Limit of Detection; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D016689:Trypanosoma vivax / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D000591:Amino Acid Isomerases; C022451:proline racemase