DIVERSITY OF TRYPANOSOMA-CRUZI STOCKS AND CLONES DERIVED FROM CHAGAS-DISEASE PATIENTS .2. ISOZYME AND RFLP CHARACTERIZATIONS

Isoenzyme and RFLP analyses were carried on freshly isolated Trypanoso ma cruzi stocks and subsequent clones derived from patients with chron ic Chagas disease. The isoenzymes separated the parasite stocks and cl ones in two groups: The stock hSLU239 (group I), isolated from a heart disease patient, showed the zymodeme 3 (Z3) profile (M. A. Miles et a l., 1977, Transactions of the Royal Society of Tropical Medicine and H ygiene 71, 217-225). The stock mSLU142 (group II), isolated from a dig estive disease (megaesophagus) patient, showed the Z2 profile. The par asite clones m1, m2, m3, and m4, derived from mSLU142, and clones h1 a nd h2, derived from hSLU239, showed isoenzyme profiles similar to thos e of 22 and ZA (Miles et al. 1977; J. A. Romanha, 1982, Thesis, Univer sidade Federal dr Minas Gerais). Furthermore, the T. cruzi clones deri ved from the cardiac disease patient differed from those derived from the megacolon patient in 3 of the 13 enzymes analyzed. RFLP analysis s howed polymorphism at the EcoRI and PstI restriction fragments of the DNA sequences coding the glycolytic enzymes ALD, GPI, GAPDH: and PYK a nd separated the T. cruzi stocks and clones in three groups: I compris ing the stock hSLU239 and clone m4, which was classified as homozygous CC, BB, AA, and AA for the ALD, GPI, PYK, and GAPDH genes, respective ly, II. formed by the parasite stock mSLU142 and clones h1 and h2 (der ived from hSLU239). which was classified as homozygous AA and BE for t he GPI and GAPDH, respectively, and classified as AC for the ALD genes ; and III, which included the clones m1, m2, and m3, (derived from mSL U142). which was classified as homozygous AA, AA, CC, and BB for ALD, GPI, PYK, and GAPDH genes. respectively. These findings show that the infection of each Chagas disease patient may be produced by geneticall y diverse mixed parasite populations. (C) 1996 Academic Press, Inc.