Serodiagnosis of bovine trypanosomosis caused by non-tsetse transmitted Trypanosoma (Duttonella) vivax parasites using the soluble form of a Trypanozoon variant surface glycoprotein antigen

Uzcanga, Graciela L.; Pérez-Rojas, Yenis; Camargo, Rocío; Izquier, Adriana; Noda, José A.; Chacín, Ronny; Parra, Nereida; Ron, Lenin; Rodríguez‐Hidalgo, Richar; Bubis, José

doi:10.1016/j.vetpar.2016.01.007

Cited by 13 publications

(4 citation statements)

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“…1. Purification of T. equiperdum sVSG and mVSG. (A) sVSG was purified as previously described (Uzcanga et al 2004, 2016), and mVSG was purified from freshly isolated T. equiperdum parasites by organic solvent extraction. Aliquots of both purified proteins were separated by SDS–PAGE and stained with Coomassie Brilliant Blue (left), or electrotransferred to a nitrocellulose filter and detected using mouse anti-sVSG polyclonal antibodies (right).…”

Section: Resultsmentioning

confidence: 99%

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Identification of potential protein partners that bind to the variant surface glycoprotein inTrypanosoma equiperdum

et al. 2017

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Trypanosoma equiperdum possesses a dense coat of a variant surface glycoprotein (VSG) that is used to evade the host immune response by a process known as antigenic variation. Soluble and membrane forms of the predominant VSG from the Venezuelan T. equiperdum TeAp-N/D1 strain (sVSG and mVSG, respectively) were purified to homogeneity; and antibodies against sVSG and mVSG were raised, isolated, and employed to produce anti-idiotypic antibodies that structurally mimic the VSG surface. Prospective VSG-binding partners were initially detected by far-Western blots, and then by immunoblots using the generated anti-idiotypic antibodies. Polypeptides of ~80 and 55 kDa were isolated when anti-idiotypic antibodies-Sepharose affinity matrixes were used as baits. Mass spectrometry sequencing yielded hits with various proteins from Trypanosoma brucei such as heat-shock protein 70, tryparedoxin peroxidase, VSG variants, expression site associated gene product 6, and two hypothetical proteins. In addition, a possible interaction with a protein homologous to the glutamic acid/alanine-rich protein from Trypanosoma congolense was also found. These results indicate that the corresponding orthologous gene products are candidates for VSG-interacting proteins in T. equiperdum.

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Section: Resultsmentioning

confidence: 99%

“…Purification of sVSG from T. equiperdum sVSG was purified from frozen TeAp-N/D1 T. equiperdum parasites as previously described (Uzcanga et al 2004(Uzcanga et al , 2016. Briefly, the parasite soluble fraction was loaded onto a Q-Sepharose column (40 mL) connected in tandem with an S-Sepharose column (15 mL).…”

Section: Parasitesmentioning

confidence: 99%

Identification of potential protein partners that bind to the variant surface glycoprotein inTrypanosoma equiperdum

et al. 2017

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“…Most studies have focused on the feeding of the vector of the parasite or on the serological detection of the parasite only (Mukhtar et al ., 2000; Singh et al ., 2008; Akter et al ., 2012). However, molecular techniques are important to determine the participation of different host animals in the cycle of the disease, as well as to identify the parasite species with which host animals are in contact, because the occurrence of other flagellated protozoa in these animals might show cross-reactions when using a serological technique for detection and diagnosis (Verlooo et al ., 2000; Luciano et al ., 2009; Eyford et al ., 2011; Syvagothi et al ., 2014; Uzcanga et al ., 2016).…”

Section: Discussionmentioning

confidence: 99%

First isolation of Leishmania infantum by blood culture in bovines from endemic area for canine visceral leishmaniasis

et al. 2019

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Leishmaniasis is considered a parasitic disease that still causes serious consequences for mankind, because it presents a high mortality rate worldwide. Considered multi-hosts, the parasites of the genus Leishmania are able of infecting a wide variety of animal species. The dog was considered the main source of infection of visceral leishmaniasis (VL), in the urban area. However, the role of other animal species in the epidemiological cycle of the disease, such as cattle, remains unclear. Therefore, the aim of the present study was to evaluate the occurrence of Leishmania spp. in 100 bovines (Bos taurus) from an area endemic for canine VL, using blood culture and molecular analysis. By the sequencing analysis, one sample showed 100% similarity with Leishmania infantum. The results provide the first case of L. infantum isolation in one bovine from the periurban areas of Bauru, state of São Paulo, Brazil.

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“…equiperdum p64, as cross-reactive diagnostic antigens for T . vivax cattle infections has been described [ 9 ] and these may lead to new diagnostic tools. Nevertheless, at the moment, farmers mostly rely upon symptom-based diagnosis, which is complicated by the numerous other diseases with similar manifestations in the endemic regions.…”

Section: Introductionmentioning

confidence: 99%

Proteomic Identification of Immunodiagnostic Antigens for Trypanosoma vivax Infections in Cattle and Generation of a Proof-of-Concept Lateral Flow Test Diagnostic Device

et al. 2016

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Trypanosoma vivax is one of the causative agents of Animal African Trypanosomosis in cattle, which is endemic in sub-Saharan Africa and transmitted primarily by the bite of the tsetse fly vector. The parasite can also be mechanically transmitted, and this has allowed its spread to South America. Diagnostics are limited for this parasite and in farm settings diagnosis is mainly symptom-based. We set out to identify, using a proteomic approach, candidate diagnostic antigens to develop into an easy to use pen-side lateral flow test device. Two related members the invariant surface glycoprotein family, TvY486_0045500 and TvY486_0019690, were selected. Segments of these antigens, lacking N-terminal signal peptides and C-terminal transmembrane domains, were expressed in E. coli. Both were developed into ELISA tests and one of them, TvY486_0045500, was developed into a lateral flow test prototype. The tests were all evaluated blind with 113 randomised serum samples, taken from 37 calves before and after infection with T. vivax or T. congolense. The TvY486_0045500 and TvY486_0019690 ELISA tests gave identical sensitivity and specificity values for T. vivax infection of 94.5% (95% CI, 86.5% to 98.5%) and 88.0% (95% CI, 75.7% to 95.5%), respectively, and the TvY486_0045500 lateral flow test prototype a sensitivity and specificity of 92.0% (95% CI, 83.4% to 97.0%) and 89.8% (95% CI, 77.8% to 96.6%), respectively. These data suggest that recombinant TvY486_0045500 shows promise for the development of a pen-side lateral flow test for the diagnosis of T. vivax animal African trypanosomosis.

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Serodiagnosis of bovine trypanosomosis caused by non-tsetse transmitted Trypanosoma (Duttonella) vivax parasites using the soluble form of a Trypanozoon variant surface glycoprotein antigen

Cited by 13 publications

References 52 publications

Identification of potential protein partners that bind to the variant surface glycoprotein inTrypanosoma equiperdum

Identification of potential protein partners that bind to the variant surface glycoprotein inTrypanosoma equiperdum

First isolation of Leishmania infantum by blood culture in bovines from endemic area for canine visceral leishmaniasis

Proteomic Identification of Immunodiagnostic Antigens for Trypanosoma vivax Infections in Cattle and Generation of a Proof-of-Concept Lateral Flow Test Diagnostic Device

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