Genotyping of Trypanosoma cruzi DTUs and Trypanosoma rangeli genetic groups in experimentally infected Rhodnius prolixus by PCR-RFLP

Sá, Amanda Regina Nichi; Dias, Greicy Brisa Malaquias; Kimoto, Karen Yuki; Steindel, Mário; Grisard, Edmundo C.; Toledo, M. J. O.; Gomes, Mônica Lúcia

doi:10.1016/j.actatropica.2016.01.006

Cited by 9 publications

(13 citation statements)

References 39 publications

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“…The Trypanosoma cruzi-triatomine interaction has been studied by several researchers over the years and although there are several reports 14,[35][36][37][38][39] , many gaps in the knowledge about the parasite cycle in invertebrate hosts exist. However, it is clear that Chagas disease depends highly on the degree of interaction between the vector and parasite 15 .…”

Section: Discussionmentioning

confidence: 99%

“…The excreta and intestinal contents of insects were stored in 70% ethanol prior to deoxyribonucleic acid (DNA) extraction 14 and confirmed for infection by molecular analysis through PCR. Triatomines that tested positive in at least one of four techniques used (FE, GC, DC, and PCR) were considered as infected.…”

Section: Parasitological and Molecular Evaluationmentioning

confidence: 99%

“…DNA extraction: Samples of the triatomine excreta and intestinal content were centrifuged at 2,500rpm for 20 min for removing the 70% ethanol and DNA was extracted using a standard phenol chloroform method 14,32 . Briefly, 500μL of lysis buffer (80mM NaCl/45mM ethylenediaminetetraacetic acid (EDTA), pH 8.0/1% sodium dodecyl sulfate) supplemented with 5μL of 10mg/mL proteinase K (Invitrogen, U.S.A.) was added and the resulting mixture was incubated at 37°C overnight.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…The natural populations of T. cruzi are polyclonal and its subpopulations can be exposed to selective pressure depending on the genetic constitution of the vertebrate host 10,11 , different species of triatomines in which the parasites develop and circulate 12,13 , and genetic constitution of these parasites 14 . However, some aspects of the coevolution of this parasite and vector bugs remain poorly understood 15 .…”

Section: Introductionmentioning

confidence: 99%

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Sympatry influence in the interaction of Trypanosoma cruzi with triatomine

Dworak

Araújo

Gomes

et al. 2017

Rev. Soc. Bras. Med. Trop.

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Introduction: Trypanosoma cruzi, the etiologic agent of Chagas disease, is widely distributed in nature, circulating between triatomine bugs and sylvatic mammals, and has large genetic diversity. Both the vector species and the genetic lineages of T. cruzi present a varied geographical distribution. This study aimed to verify the influence of sympatry in the interaction of T. cruzi with triatomines. Methods: The behavior of the strains PR2256 (T. cruzi II) and AM14 (T. cruzi IV) was studied in Triatoma sordida (TS) and Rhodnius robustus (RR). Eleven fifth-stage nymphs were fed by artificial xenodiagnosis with 5.6 × 10 3 blood trypomastigotes/0.1mL of each T. cruzi strain. Every 20 days, their excreta were examined for up to 100 days, and every 30 days, the intestinal content was examined for up to 120 days, by parasitological (fresh examination and differential count with Giemsa-stained smears) and molecular (PCR) methods. Rates of infectivity, metacyclogenesis and mortality, and mean number of parasites per insect and of excreted parasites were determined. Results: Sympatric groups RR+AM14 and TS+PR2256 showed higher values of the four parameters, except for mortality rate, which was higher (27.3%) in the TS+AM14 group. General infectivity was 72.7%, which was mainly proven by PCR, showing the following decreasing order: RR+AM14 (100%), TS+PR2256 (81.8%), RR+PR2256 (72.7%) and TS+AM14 (36.4%). Conclusions: Our working hypothesis was confirmed once higher infectivity and vector capacity (flagellate production and elimination of infective metacyclic forms) were recorded in the groups that contained sympatric T. cruzi lineages and triatomine species.

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Section: Discussionmentioning

confidence: 99%

Section: Parasitological and Molecular Evaluationmentioning

confidence: 99%

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Sympatry influence in the interaction of Trypanosoma cruzi with triatomine

Dworak

Araújo

Gomes

et al. 2017

Rev. Soc. Bras. Med. Trop.

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“…For XC, an aliquot of 250 μL of the IC was inoculated into 2.5 mL of LIT (Liver infusion tryptose) culture medium, with and without antibiotic (6.6 mg/mL of penicillin) (Bronfen et al, 1989;Bisugo et al, 1998). Finally, the remaining IC was added to 500 µL of 70 °GL ethyl alcohol and stored at -20 ºC until DNA extraction (Sá et al, 2013;Sá et al, 2016). The remaining 10 specimens of P. megistus from Janiópolis were received without life and were stored in bottles containing a 70 °GL ethyl alcohol solution, not allowing the performance of FE and XC.…”

Section: Analysis Of Excreta and Intestinal Contentmentioning

confidence: 99%

Investigation regarding the presence of Trypanosoma cruzi in triatomines and humans in rural households in the State of Paraná, southern Brazil

Abreu¹,

Silva²,

Sarto³

et al. 2021

RSD

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Trypanosoma cruzi, the etiologic agent of Chagas disease (CD), is transmitted by hematophagous insects belonging to the subfamily Triatominae. After elimination of Triatoma infestans, the infestation of human dwellings by secondary species of vectors continues to pose a risk of transmission of the parasite. Our aim was to investigate the T. cruzi presence in triatomines and humans in rural households in the State of Paraná, southern Brazil. The capture of the insects was carried out by technicians of the municipalities after residents reported the outbreak. Five residents and 27 triatomines captured in four municipalities in the North and Midwest of the state were evaluated. The research of T. cruzi was carried out using parasitological, serological, and molecular techniques, in human blood, excreta, intestinal contents and insect macerate. Panstrongylus megistus, P. geniculatus and Triatoma sp. were identified. Ten specimens of P. megistus were captured in a house in Mandaguari with five residents and presented an infection rate of 70% for T. cruzi like. All residents tested negative for T. cruzi infection. Another 15 P. megistus were captured in the peridomicile in Janiópolis and had 100.0% positivity. The only adult specimen of P. geniculatus captured in the intradomicile in Amaporã, as well as the nymph of Triatoma in the peridomicile in Paiçandu, were negative. The finding of P. megistus naturally infected by T. cruzi in households in rural area of Paraná demonstrates a potential risk of vector transmission of CD in these regions.

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Limit of detection of PCR/RFLP analysis of cytochrome oxidase II for the identification of genetic groups of Trypanosoma cruzi and Trypanosoma rangeli in biological material from vertebrate hosts

et al. 2018

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Mixed infections with Trypanosoma cruzi and Trypanosoma rangeli and their different genetic groups occur frequently in vertebrate hosts and are difficult to detect by serology. In the present study, we evaluated the limit of detection of polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis of cytochrome oxidase II (COII) for the identification of genetic groups of these two parasites in blood and tissue from vertebrate hosts. Reconstitution experiments were performed using human blood (TcI/TcII and KP1+/KP1-) and mouse tissue (TcI/TcII). We tested blood from patients who were in the chronic phase of Chagas disease and tissue from animals that were experimentally infected with all possible combinations of six discrete typing units. In blood samples, T. cruzi and T. rangeli were detected when 5 parasites (pa) were present in the sample, and genetic groups were identified when at least 50 pa were present in the sample. T. cruzi alone could be detected with 1 pa and genotyped (TcI/TcII) with 2 pa. T. rangeli was detected with 2 pa and genotyped (KP+/KP1-) with 25 pa. The present method more readily detected TcII and KP1- in both admixtures and alone. In mouse tissue, TcI and TcII were detected with at least 25 pa. The analysis of blood samples from patients and tissue from animals that were experimentally infected revealed low parasite loads in these hosts, which were below the limit of detection of the present method and could not be genotyped. Our findings indicate that the performance of PCR/RFLP analysis of COII is directly related to the amount and proportion of parasites that are present in the sample and the genetic groups to which the parasites belong.

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Genotyping of Trypanosoma cruzi DTUs and Trypanosoma rangeli genetic groups in experimentally infected Rhodnius prolixus by PCR-RFLP

Cited by 9 publications

References 39 publications

Sympatry influence in the interaction of Trypanosoma cruzi with triatomine

Sympatry influence in the interaction of Trypanosoma cruzi with triatomine

Investigation regarding the presence of Trypanosoma cruzi in triatomines and humans in rural households in the State of Paraná, southern Brazil

Limit of detection of PCR/RFLP analysis of cytochrome oxidase II for the identification of genetic groups of Trypanosoma cruzi and Trypanosoma rangeli in biological material from vertebrate hosts

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