Internal transcribed spacers (ITS) of Trypanosoma rangeli ribosomal DNA (rDNA): a useful marker for inter-specific differentiation

Beltrame-Botelho, I.T.; Gaspar-Silva, Daniela; Steindel, Mário; Dávila, Alberto M. R.; Grisard, Edmundo C.

doi:10.1016/j.meegid.2004.05.007

Cited by 31 publications

(15 citation statements)

References 36 publications

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“…In fact, this marker has been very useful in molecular analysis to establish relationships among and within trypanosome species such as T. b. brucei, T. b. gambiense, T. cruzi, T. rangeli, and T. evansi (Agbo et al 2001, Santos et al 2002, Beltrame-Botelho et al 2005, Khuchareontaworn et al 2007, Areekit et al 2008 *Corresponding author: wlrivera@science.upd.edu.ph Trypanosoma evansi isolates from the Philippines 7 2011). Previous study based on ITS regions of T. brucei has established genotypic discrimination between its two subspecies (Agbo et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Villareal

Mingala

Rivera³

2013

Acta Parasitologica

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Trypanosoma evansi infection in the Philippines is frequently reported to affect the country's livestock, particularly, the buffaloes. To assess the prevalence and intraspecific diversity of T. evansi in the country, blood samples from water buffaloes in different geographical regions were collected during an outbreak. T. evansi was detected in all 79 animals tested using PCR targeting the RoTat 1.2 VSG gene. Sequencing of the rDNA complete internal transcribed spacer (ITS) region including the 5.8S subunit showed high similarity (99-100%) between Philippine isolates and known T. evansi isolates in Genbank. Tree construction based on the same region confirmed the close relationship between Philippine and reported Thai isolates as compared to Egyptian isolates separated by relatively small genetic distances, 47 polymorphisms, despite the clustering in four branches. Overall, the results of this study prove genetic diversity within T. evansi species despite previous reports on limited heterogeneity among isolates worldwide.

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Section: Introductionmentioning

confidence: 99%

Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Villareal

Mingala

Rivera³

2013

Acta Parasitologica

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“…It has often been reported that the ITS regions of the rRNA gene are often highly variable with respect to nucleotide composition and that this characteristic can be used to distinguish both morphologically distinct fungal species and strains of the same fungal species (Beltrame-Botelho et al 2005;Healy et al 2004;Tanabe et al 2004). As such, the ITS region of the rRNA gene is generally believed to represent a convenient target for the molecular identification of specific species of fungi.…”

Section: Discussionmentioning

confidence: 99%

“…These apparent strain differences are often not stable upon subculture, and they may simply be artifacts due to specific growthcondition requirements for the colony or may arise solely due to the presence of contaminating bacteria. It has been suggested that polymorphism analysis of internal transcribed spacer (ITS) regions of the ribosomal RNA gene is a valuable technique for both strain typing and species identification for a number of important fungi (Beltrame-Botelho et al 2005;Healy et al 2004;Tanabe et al 2004). However, little use has been made of this technique in the study of phylogeny of genus Antrodia.…”

Section: Introductionmentioning

confidence: 99%

Phylogenetic analysis of Antrodia species and Antrodia camphorata inferred from internal transcribed spacer region

Chiu

2006

Antonie van Leeuwenhoek

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The species of Antrodia are one of the difficult-to-classify and obscure groups of poroid Aphyllophorales based on morphological appearance. However, it is becoming increasingly important to reliably identify the entire suite of Antrodia camphorata strains and Antrodia species due to the potential pharmaceutical value of their biologically active ingredients. In this study, the internal transcribed spacer (ITS) region of the ribosomal RNA gene (rDNA) was sequenced and phylogenetically analyzed in a number of Antrodia fungal species and strains. ITS amplicons from the Antrodia species tested ranged in size from 543 to 610 bp; the size of the ITS of A. camphorata strains ranged from 592 to 596 bp. The overall sizes of ITS2 and 5.8S ribosomal RNA gene of all A. camphorata strains tested in this study were shown to be 217 and 158 bp, respectively. A phylogenetic analysis of ITS data generated, which included sequences of 11 A. camphorata strains and nine other Antrodia species, showed three clearly distinct groups. Group 1 includes A. camphorata, Antrodia salmonea, and Antrodia carbinca strains. Within Group 2, Antrodia sinuosa and Antrodia xantha were clustered together. Group 3 contained Antrodia albida, A. heteromorpha, A. serialis, and A. malicola. The observed sequence diversity among ITS alleles provided an effective tool for differentiating strains of A. camphorata, A. salmonea, A. xantha, A. sinuosa, or A. serialis. Polymorphisms arising within the ITS1-5.8S-ITS2 region can provide practical markers for establishing a foundation for the further expansion of an ITS sequence database of medically important fungi.

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“…Accordingly, single or mixed infections with these species have been described (D'Alessandro 1976; D'Alessandro-Bacigalupo and Gore-Saravia 1992; Cuba-Cuba 1998;Guhl and Vallejo 2003;Sousa et al 2008a), being of utmost importance their accurate differential diagnosis. Classical parasitological and immunological techniques, besides several biochemical and molecular analyses (mainly based on polymerase chain reaction products) have been used for characterization and distinction between T. rangeli and T. cruzi (e.g., Miles et al 1983;Guhl et al 1985Guhl et al , 2002Holguín et al 1987;Steindel et al 1991Steindel et al , 1994Grisard et al 1999a, b;Toaldo et al 2001;Maia-da-Silva et al 2004;Santa-Izabel et al 2004;Beltrame-Botelho et al 2005;Sousa et al 2008a;Faissal et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Variable sensitivity to complement-mediated lysis among Trypanosoma rangeli reference strains

2011

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Six reference strains of Trypanosoma rangeli from different days of growth in axenic cultures were assayed for susceptibility to complement-mediated lysis by non-immune guinea-pig serum. Their authenticity was also confirmed by isoenzyme analyses. Parasites were incubated with 25% active or 68°C-inactivated serum (37°C, 30 min) for all tests; thereafter the lysis rates were determined. Highly variable lysis percentages were observed among T. rangeli strains and in the same stock at different growing days. In a few assays, three strains (Macias, R-1625 and Choachi) presented total or very high resistance. The others (H-14, San Agustín and SC-58) were generally most susceptible, and could reach lysis rates as high as Trypanosoma cruzi. After incubation with active sera, the epimastigotes were usually the predominant stages, being followed by spheromastigotes and/or transitional forms. Those stages and trypomastigotes could also be partially susceptible. In four strains, the short epimastigotes were more resistant to lysis than the long ones. Experiments with C3-deficient serum displayed total or partial participation of the alternative-complement pathway in T. rangeli lysis. This study confirmed the variable complement sensitivity of T. rangeli, which can be related to its intraspecific heterogeneity, to the remarkable complexity of its life-cycle stages, and to the methodology employed.

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Internal transcribed spacers (ITS) of Trypanosoma rangeli ribosomal DNA (rDNA): a useful marker for inter-specific differentiation

Cited by 31 publications

References 36 publications

Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Phylogenetic analysis of Antrodia species and Antrodia camphorata inferred from internal transcribed spacer region

Variable sensitivity to complement-mediated lysis among Trypanosoma rangeli reference strains

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