A Trypanosoma cruzi Genome Tandem Repetitive Satellite DNA Sequence as a Molecular Marker for a LAMP Assay for Diagnosing Chagas' Disease.

A Trypanosoma cruzi Genome Tandem Repetitive Satellite DNA Sequence as a Molecular Marker for a LAMP Assay for Diagnosing Chagas' Disease. Dis Markers. 2020;2020:8074314 Authors: Ordóñez D, Fernández-Soto P, Fernández-Martín AM, Crego-Vicente B, Febrer-Sendra B, Diego JG, Vicente B, López-Abán J, Belhassen-García M, Muro A, Patarroyo MA Abstract Chagas' disease is a neglected tropical disease caused by Trypanosoma cruzi which is endemic throughout Latin America and is spread by worldwide migration. Diagnosis is currently limited to serological and molecular techniques having variations regarding their sensitivity and specificity. This work was aimed at developing a new sensitive, applicable, and cost-effective molecular diagnosis technique for loop-mediated isothermal amplification-based detection of T. cruzi (Tc-LAMP). The results led to determining a highly homologous satellite repeat region (231 bp) among parasite strains as a molecular marker for diagnosing the disease. Tc-LAMP was performed correctly for detecting parasite DNA (5 fg for the CL Brener strain and 50 fg for the DM28, TcVI, and TcI strains). Assay results proved negative for DNA from 16 helminth species and 7 protozoa, including Leishmania spp. Tc-LAMP based on the highly repeated T. cruzi satellite region is thus proposed as an important alternative for diagnosing T. cruzi infection, overcoming other methods' l...
Source: Disease Markers - Category: Laboratory Medicine Tags: Dis Markers Source Type: research

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CONCLUSION: These results suggest that the double or triple infection is a phenomenon existing in almost all the coendemics areas and mammals studied, which might influence the mechanisms of adaptation and pathogenicity of these parasites. PMID: 32655075 [PubMed - in process]
Source: Journal of Vector Borne Diseases - Category: Infectious Diseases Tags: J Vector Borne Dis Source Type: research
This study was carried out in accordance with the recommendations of Institutional Review Board of the Federal University of Bahia, Brazil, with written informed consent from all subjects. All subjects gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the Institutional Review Board of the Federal University of Bahia, Brazil (approval number 693.111). Human Blood Samples and Preparation of Peripheral Blood Cells Peripheral blood mononuclear cells (PBMC) were separated from heparinized venous blood by Ficoll-Hypaque gradient centrifugation. Cells were then washed in ...
Source: Frontiers in cellular and infection microbiology - Category: Microbiology Source Type: research
This study was performed in accordance with the ethical standards of the Institutional Animal Care and Use Committee (IACUC, approval FUA-007-14) from the Unidad de Biología Comparativa (UBA) at Pontificia Universidad Javeriana (PUJ, Bogotá, Colombia). All animal studies were conducted in accordance with the “Guide for the Care and Use of Laboratory Animals” from UBA-PUJ. The present study was described according to the Animal Research: Reporting in vivo Experiments (ARRIVE) criteria from the National Center for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (32). Mice ...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
Abstract The choice of cost-effective molecular methods for diagnosing and monitoring of Chagas disease before and after treatment is crucial in endemic countries with high patients' demand and limited financial resources. To this end, a kDNA was compared to a satellite real-time quantitative PCR (sat-qPCR), both amplifications using Sybr Green instead of Taqman hydrolysis probes. Non-isogenic Swiss albino mice were infected with a small inoculum of the highly virulent and partially resistant to benznidazole Y strain, belonging to T. cruzi discrete typing unit II (DTU-II) that predominates in Atlantic and Central ...
Source: Experimental Parasitology - Category: Parasitology Authors: Tags: Exp Parasitol Source Type: research
This study aimed to evaluate the performance of flow cytometry with fixed parasites for VL diagnosis, comparing it with four other serological tests. Samples from two endemic VL regions in Brazil, diagnosed by direct examination (DG1) and by at least two or one standard serological test (DG2 and DG3, respectively), as well as patients with chronic Chagas' disease (CG1) and healthy controls (CG2) were used in this study. The flow cytometry results were expressed as levels of IgG reactivity, based on the percentage of positive fluorescent parasites (PPFP). Using a 1:4096 serum dilution, a ROC curve analysis of the serum titr...
Source: Journal of Immunological Methods - Category: Allergy & Immunology Authors: Tags: J Immunol Methods Source Type: research
Abstract Neglected Tropical Diseases (NTDs) comprise of a group of seventeen infectious conditions endemic in many developing countries. Among these diseases are three of protozoan origin, namely leishmaniasis, Chagas disease, and African trypanosomiasis, caused by the parasites Leishmania spp., Trypanosoma cruzi, and Trypanosoma brucei respectively. These diseases have their own unique challenges which are associated with the development of effective prevention and treatment methods. Collectively, these parasitic diseases cause more deaths worldwide than all other NTDs combined. Moreover, many current therapies for these ...
Source: Journal of Venomous Animals and Toxins including Tropical Diseases - Category: Tropical Medicine Source Type: research
Conclusion Although NTDs may increasingly pose a threat to health in the U.S., insufficient data exists to ascertain the prevalence and geographic distribution of risk factors relevant to these diseases. Concern over the potential for increasing rates of NTD infection and transmission in Texas led the Texas State Legislature to mandate the establishment of a surveillance program for NTDs. We tested one potential methodology for the surveillance of NTDs; the use of CASPER for assessing the prevalence of risk factors in Brazos County, Texas. Although Texas has a population of more than 25 million, the second largest in the U...
Source: PLOS Currents Outbreaks - Category: Epidemiology Authors: Source Type: research
Chagas disease, caused by Trypanosoma cruzi, although endemic in many parts of Central and South America, is emerging as a global health threat through the potential contamination of blood supplies. Consequently, in the absence of a gold standard assay for the diagnosis of Chagas disease, additional antigens or strategies are needed. A proteomic analysis of the trypomastigote excreted-secreted antigens (TESA) associated with exosomal vesicles shed by T. cruzi identified ~80 parasite proteins, with the majority being trans-sialidases. Mass spectrometry analysis of immunoprecipitation products performed using Chagas immune s...
Source: Journal of Clinical Microbiology - Category: Microbiology Authors: Tags: Immunoassays Source Type: research
PREVALENCE OF AMERICAN TRYPANOSOMIASIS AND LEISHMANIASES IN DOMESTIC DOGS IN A RURAL AREA OF THE MUNICIPALITY OF SÃO JOÃO DO PIAUÍ, PIAUÍ STATE, BRAZIL. Rev Inst Med Trop Sao Paulo. 2016 Nov 03;58:79 Authors: Perez TD, Figueiredo FB, Junior AA, Silva VL, Madeira MF, Brazil RP, Coura JR Abstract Chagas disease and the leishmaniases are endemic zoonoses of great importance to public health in the state of Piauí, Brazil. The domestic dog (Canis familiaris) is a major reservoir, host of Trypanosoma cruzi and Leishmania spp. in both urban and rural areas, playing an impor...
Source: Revista do Instituto de Medicina Tropical de Sao Paulo - Category: Tropical Medicine Authors: Tags: Rev Inst Med Trop Sao Paulo Source Type: research
by Geraldo Pedral-Sampaio, Jess é S. Alves, Albert Schriefer, Andréa Magalhães, Roberto Meyer, Marshall J. Glesby, Edgar M. Carvalho, Lucas P. Carvalho Diagnosis of cutaneous leishmaniasis (CL) relies on clinical presentation, parasite isolation, histopathologic evaluation and positive Montenegro skin test. However, the low amounts of parasites in the lesion of these individuals make parasite isolation and histopatologic diagnosis unreliable, oft en leading to false-negative results. Also, 15% of people living in endemic areas have sub-clinical infection characterized by positive Montenegro skin test,...
Source: PLoS One - Category: Biomedical Science Authors: Source Type: research
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