This site needs JavaScript to work properly. Please enable it to take advantage of the complete set of features!
Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log in
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 Mar;12(3):196-205.
doi: 10.1089/vbz.2011.0683. Epub 2011 Oct 24.

Trypanosoma cruzi genotypes of insect vectors and patients with Chagas of Chile studied by means of cytochrome b gene sequencing, minicircle hybridization, and nuclear gene polymorphisms

Affiliations

Trypanosoma cruzi genotypes of insect vectors and patients with Chagas of Chile studied by means of cytochrome b gene sequencing, minicircle hybridization, and nuclear gene polymorphisms

Marco Arenas et al. Vector Borne Zoonotic Dis. 2012 Mar.

Abstract

Fifty-six Trypanosoma cruzi stocks from Chile and neighboring countries and different hosts, humans, and Triatoma infestans and Mepraia sp., vectors of domiciliary and natural environments were characterized by using three molecular markers. These were cytochrome b (Cyt b) gene sequencing, minicircle DNA blotting, and hybridization with five genotype-specific DNA probes and nuclear analysis of 1f8 and gp72 by polymerase chain reaction-restriction fragment length polymorphism. The results with all three molecular markers are concordant, with minor limitations, grouping T. cruzi stocks into four discrete typing units (DTUs) (TcI, TcII, TcV, and TcVI). TcI and TcII stocks were heterogeneous. TcI and TcII stocks were clustered in two main subgroups determined by Cyt b gene sequencing and minicircle hybridization. However, TcV and TcVI stocks were homogeneous and not differentiated by Cyt b gene sequencing or minicircle DNA hybridization. The discriminatory power and limitations of the molecular markers are discussed, as well as the distribution of the four DTUs in the domiciliary and sylvatic transmission cycles of Chile and the limitations of each marker for molecular epidemiological studies performed with T. cruzi stocks rather than the analysis of direct T. cruzi samples from natural hosts.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Maximum likelihood phylogenetic tree of Trypanosoma cruzi strains based on Cyt b gene sequences inferred with the model GTR+G (G=0.1130, -lnL=3851.9282). Numbers at nodes are the support values derived from 1000 replicates. TcI, TcII, and TcV-TcVI are the three clusters supported by significant bootstrap values representing the genotypes of T. cruzi. A, B, subgroups within each cluster.
FIG. 2.
FIG. 2.
Representative results of hybridization patterns of different T. cruzi stocks belonging to TcI and other discrete typing units (DTUs). (A, C) Ethidium bromide staining of a minicircle polymerase chain reaction (PCR) product. (B, D) Hybridization with the TcI probe (sp104cl1). M, molecular marker.
FIG. 3.
FIG. 3.
Hybridization patterns of different T. cruzi stocks and clones belonging to DTU TcII and other DTUs. (A) Ethidium bromide staining from minicircle PCR products. (B) Hybridization with the TcII probe (CBBcl3). (C) Hybridization with the TcII probe (IVVcl4). (D) Hybridization with the TcV probe (NRcl3).
FIG. 4.
FIG. 4.
Representative results of hybridization patterns of different T. cruzi stocks belonging to different DTUs. (A) Ethidium bromide staining of a minicircle PCR product. (B) Hybridization with the TcI probe (sp104cl1). (C) Hybridization with the TcII probe (CBBcl3). (D) Hybridization with the TcIV probe (IVVcl4). (E) Hybridization with the TcV probe (NRcl3). (F) Hybridization with the TcVI probe (v195cl1). M, molecular weight marker.
FIG. 5.
FIG. 5.
Hybridization patterns of different T. cruzi clones belonging to different DTUs from different countries. (A) Ethidium bromide staining of a minicircle PCR amplification product. (B) Hybridization with the TcV probe (NRcl3). (C) Hybridization with the TcVI probe (v195cl1).
FIG. 6.
FIG. 6.
PCR–restriction fragment length polymorphism patterns of representative T. cruzi stocks belonging to different DTUs: sp54 (TcI); T and MxCh88 (TcII); M5631 (TcIII); CanIII (TcIV); Xd96 (TcV); and CH2 (TcVI). Ten or 15 μL of amplified PCR digested with the endonuclease were analyzed on a 3% agarose gel and stained with ethidium bromide. M, molecular weight marker.

References

    1. Akaike H. A new look at the statistical model identification. IEEE Trans Automat. Contr. 1974;19:716–723.
    1. Anonymous. Recommendations from a satellite meeting. Mem Inst Oswaldo Cruz. 1999;194:429–432. - PubMed
    1. Avila HA. Simpson L. Organization and complexity of minicircle-encoded guide RNAs in Trypanosoma cruzi. RNA. 1995;1:939–947. - PMC - PubMed
    1. Barnabé C. Neubauer K. Solari A. Tibayrenc M. Trypanosoma cruzi: presence of the two major phylogenetic lineages and of several lesser discrete typing units (DTUs) in Chile and Paraguay. Acta Trop. 2001;78:127–137. - PubMed
    1. Bosseno MF. Yacsik N. Vargas F. Brenière SF. Selection of Trypanosoma cruzi clonal genotypes (clonet 20 and 39) isolated from Bolivian triatomines following subculture in liquid medium. Mem Inst Oswaldo Cruz. 2000;95:601–607. - PubMed

Publication types

MeSH terms

Full text links
Atypon full text link Atypon Free PMC article
Cite

AltStyle によって変換されたページ (->オリジナル) /