Document Detail


Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites.
MedLine Citation:
PMID:  18154957     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The investigation of the importance of the genetics of Trypanosoma cruzi in determining the clinical course of Chagas disease will depend on precise characterisation of the parasites present in the tissue lesions. This can be adequately accomplished by the use of hypervariable nuclear markers such as microsatellites. However the unilocal nature of these loci and the scarcity of parasites in chronic lesions make it necessary to use high sensitivity PCR with nested primers, whose design depends on the availability of long flanking regions, a feature not hitherto available for any known T. cruzi microsatellites. Herein, making use of the extensive T. cruzi genome sequence now available and using the Tandem Repeats Finder software, it was possible to identify and characterise seven new microsatellite loci--six composed of trinucleotide (TcTAC15, TcTAT20, TcAAT8, TcATT14, TcGAG10 and TcCAA10) and one composed of tetranucleotide (TcAAAT6) motifs. All except the TcCAA10 locus were physically mapped onto distinct intergenic regions of chromosome III of the CL Brener clone contigs. The TcCAA10 locus was localised within a hypothetical protein gene in the T. cruzi genome. All microsatellites were polymorphic and useful for T. cruzi genetic variability studies. Using the TcTAC15 locus it was possible to separate the strains belonging to the T. cruzi I lineage (DTU I) from those belonging to T. cruzi II (DTU IIb), T. cruzi III (DTU IIc) and a hybrid group (DTU IId, IIe). The long flanking regions of these novel microsatellites allowed construction of nested primers and the use of full nested PCR protocols. This strategy enabled us to detect and differentiate T. cruzi strains directly in clinical specimens including heart, blood, CSF and skin tissues from patients in the acute and chronic phases of Chagas disease.
Authors:
Helder Magno Silva Valadares; Juliana Ramos Pimenta; Jorge Marcelo de Freitas; Tomás Duffy; Daniella C Bartholomeu; Riva de Paula Oliveira; Egler Chiari; Maria da Consolação Vieira Moreira; Geraldo Brasileiro Filho; Alejandro Gabriel Schijman; Glória Regina Franco; Carlos Renato Machado; Sérgio Danilo Junho Pena; Andréa Mara Macedo
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-11-07
Journal Detail:
Title:  International journal for parasitology     Volume:  38     ISSN:  0020-7519     ISO Abbreviation:  Int. J. Parasitol.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-04-21     Completed Date:  2008-12-04     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0314024     Medline TA:  Int J Parasitol     Country:  England    
Other Details:
Languages:  eng     Pagination:  839-50     Citation Subset:  IM    
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, Brazil.
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MeSH Terms
Descriptor/Qualifier:
Animals
Chagas Disease / genetics*,  parasitology
Chromosome Mapping
Chronic Disease
DNA Primers / genetics
Electrophoresis, Polyacrylamide Gel
Female
Gene Frequency
Genes, Protozoan*
Heart / parasitology
Humans
Male
Mice
Mice, Inbred BALB C
Microsatellite Repeats*
Parasitemia
Polymerase Chain Reaction
Polymorphism, Genetic
Pregnancy
Rectum / parasitology
Sequence Alignment
Skin / parasitology
Trypanosoma cruzi / genetics*,  isolation & purification
Chemical
Reg. No./Substance:
0/DNA Primers

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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