Article
Schizodeme analysis of Trypanosoma cruzi stocks from South and Central America by analysis of PCR-amplified minicircle variable region sequences
Registro en:
AVILA, Herbert et al. Schizodeme analysis of Trypanosoma cruzi stocks from South and Central America by analysis of PCR-amplified minicircle variable region sequences. Molecular and Biochemical Parasitology, v. 42, p. 175-188, 1990.
0166-6851
10.1016/0166-6851(90)90160-N
1872-9428
Autor
Avila, Herbert
Gonçalves, Antonio M.
Nehme, Nedia Saad
Morel, Carlos Medicis
Simpson, Larry
Resumen
Kinetoplast DNA (kDNA) was isolated from 56 stocks of Trypanosoma cruzi isolated from human patients, animals and insects
from Brazil, Venezuela, Colombia and Costa Rica. Comparison of the patterns of digested kDNA on acrylamide gels led to
the grouping of several stocks into two schizodemes. Schizodeme analysis was also performed using a set of 330-bp fragments
representing all the variable regions of the minicircle DNA molecules, which were obtained by PCR amplification of the kDNA
using conserved region primers. The results of this analysis were consistent with the analysis using total kDNA, but the more
informative restriction profiles allowed the construction of additional schizodemes. In addition, two oligomers were generated from
variable region sequences of cloned minicircles from a Y and a CI strain, and these were used as schizodeme-specific probes
to detect homologous sequences in the amplified minicircle DNAs. The results indicate that a combination of restriction enzyme
fingerprinting and hybridization of amplified variable region minicircle DNA with schizodeme-specific probes can be used for both
sensitive detection and classification of T. cruzi.