Characterization of a candidate Trypanosoma rangeli small nucleolar RNA gene and its application in a PCR-based parasite detection

Morales, Liliana; Romero, Ibeth; Díez, Hugo; Portillo, Patricia Del; Montilla, Marleny; Nicholls, Santiago; Puerta, Concepción J.

doi:10.1016/s0014-4894(03)00027-4

Cited by 22 publications

(25 citation statements)

References 51 publications

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“…Liang et al 2002;Li et al 2003). One or the other of these two Nm residues is present in other eukaryotes, but in each case the corresponding snoRNA does not have the potential to target the other site (Lowe and Eddy 1999;Xu et al 2001;Higa et al 2002;Morales et al 2002).…”

Section: Cbf5p-associated Rnas In Euglena Gracilismentioning

confidence: 99%

Pseudouridine-guide RNAs and other Cbf5p-associated RNAs in Euglena gracilis

Russell¹,

Schnare²,

Gray³

2004

RNA

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In eukaryotes, box H/ACA small nucleolar RNAs (snoRNAs) guide sites of pseudouridine (⌿) formation in rRNA. These snoRNAs reside in RNP complexes containing the putative ⌿ synthase, Cbf5p. In this study we have identified Cbf5p-associated RNAs in Euglena gracilis, an early diverging eukaryote, by immunoprecipitating Cbf5p-containing complexes from cellular extracts. We characterized one box H/ACA-like RNA which, however, does not appear to guide ⌿ formation in rRNA. We also identified four single ⌿-guide box AGA RNAs. We determined target sites for these putative ⌿-guide RNAs and confirmed that the predicted ⌿ modifications do, in fact, occur at these positions in Euglena rRNA. The Cbf5p-associated snoRNAs appear to be encoded by multicopy genes, some of which are clustered in the genome together with methylation-guide snoRNA genes. These modification-guide snoRNAs and snoRNA genes are the first ones to be reported in euglenid protists, the evolutionary sister group to the kinetoplastid protozoa. Unexpectedly, we also found and have partially characterized a selenocysteine tRNA homolog in the anti-Cbf5p-immunoprecipitated sample.

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Section: Cbf5p-associated Rnas In Euglena Gracilismentioning

confidence: 99%

Pseudouridine-guide RNAs and other Cbf5p-associated RNAs in Euglena gracilis

Russell¹,

Schnare²,

Gray³

2004

RNA

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“…Interestingly, ELIAS et al (2003) 10 and SCHIJMAN et al (2004) 26 reported a nested PCR based on SIRE which is able of detecting parasite DNA in the heart tissue of chronic chagasic patients. On the other hand, the TrF/R2 PCR, based on the repetitive genes encoding for the sno-RNA-Cl1, is a specific test that amplify a product of the same size in both T. rangeli KP1(+) and KP1(-) strains and does not present any amplification signal either with the DNA from both T. cruzi and DNA from human, mouse or even other trypanosomatids 18 . In this work, with this PCR T. rangeli was detected in a specific way in artificial infections from 2 x 10 6 parasites.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, it is necessary to develop new techniques as better options to specifically identify each of these parasite species in mixed infections. In this study we aimed to determine the capacity of two previously standardized PCR tests to detect specifically T. rangeli and T. cruzi based on the small nucleolar RNA-C11 (sno-RNA-C11) gene and the SIRE (short interspersed repetitive element) sequence inserted into the histone H2A gene, respectively 18,20 .…”

Section: Introductionmentioning

confidence: 99%

Detection of Trypanosoma cruzi and Trypanosoma rangeli infection in triatomine vectors by amplification of the histone H2A/SIRE and the sno-RNA-C11 genes

Pavía

Vallejo

Montilla

et al. 2007

Rev. Inst. Med. trop. S. Paulo

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SUMMARYTrypanosoma rangeli is non pathogenic for humans but of important medical and epidemiological interest because it shares vertebrate hosts, insect vectors, reservoirs and geographic areas with T. cruzi, the etiological agent of Chagas disease. Therefore, in this work, we set up two PCR reactions, TcH2AF/R and TrFR2, to distinguish T. cruzi from T. rangeli in mixed infections of vectors based on amplification of the histone H2A/SIRE and the small nucleolar RNA Cl1 genes, respectively. Both PCRs were able to appropriately detect all T. cruzi or T. rangeli experimentally infected-triatomines, as well as the S35/S36 PCR which amplifies the variable region of minicircle kDNA of T. cruzi. In mixed infections, whereas T. cruzi DNA was amplified in 100% of samples with TcH2AF/R and S35/S36 PCRs, T. rangeli was detected in 71% with TrF/R2 and in 6% with S35/S36. In a group of Rhodnius colombiensis collected from Coyaima (Colombia), T. cruzi was identified in 100% with both PCRs and T. rangeli in 14% with TrF/R2 and 10% with S35/S36 PCR. These results show that TcH2AF/R and TrF/R2 PCRs which are capable of recognizing all T. cruzi and T. rangeli strains and lineages could be useful for diagnosis as well as for epidemiological field studies of T. cruzi and T. rangeli vector infections.

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“…Se trabajó con las cepas H14 (MHOM/Hond/H14), Choachí (IRHO/CO/86/Choachí) y Tre de T. rangeli y las cepas Shubacbarina (IRHO/CO/95/ Shubacbarina) y Munantá (IRHO/CO/85/Munantá) de T. cruzi (18,19). Estas cepas se caracterizaron mediante su patrón de isoenzimas (20) y la amplificación de los minicírculos con los oligonucleótidos S35/S36/KP1L (2).…”

Section: Materiales Y Métodos Parásitosunclassified