Characterisation of a novel interspersed Toxoplasma gondii DNA repeat with potential uses for PCR diagnosis and PCR-RFLP analysis

PABLO C. ECHEVERRIA,; PAOLA A. ROJAS,; VALENTINA MARTIN,; EDUARDO A. GUARNERA,; VIVIANA PSZENNY,; SERGIO O. ANGEL

FEMS MICROBIOLOGY LETTERS

WILEY-BLACKWELL PUBLISHING, INC

Año: 2000 vol. 184 p. 23 - 27

0378-1097

A novel Toxoplasma gondii interspersed repeat element (TgIRE), present in most of the tachyzoite chromosomes, was characterised. Tworegions on the TgIRE sequence showed high identity to two different T. gondii expressed sequence tag cDNAs of unknown function, whichseems to be TgIRE pseudogenes. Two set of primers were designed, 2-2P and 2-3, that amplify products of 1.02 and 0.62 kb, respectively.T. gondii DNA from RH and Me49 strains was amplified with TgIRE 2-2P primers, and the respective 1.02 kb products were digested withseveral endonucleases. Different fragment patterns by gel electrophoresis were found only with MboI. Sensitivity analysis revealed that theset 2-3 was more sensitive than 2-2P, detecting by gel visualisation the amount of DNA equivalent to 1 and 10 parasites,respectively. ß 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.