دورية أكاديمية
The Development of a One-Step RT-qPCR for the Detection and Quantification of Viable Forms of Trypanosoma cruzi in Açai Samples from Areas at Risk of Chagas Disease through Oral Transmission.
| العنوان: | The Development of a One-Step RT-qPCR for the Detection and Quantification of Viable Forms of Trypanosoma cruzi in Açai Samples from Areas at Risk of Chagas Disease through Oral Transmission. |
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| المؤلفون: | Faier-Pereira A; Laboratory of Molecular Virology and Parasitology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro 21040-360, Brazil., Finamore-Araujo P; Laboratory of Molecular Virology and Parasitology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro 21040-360, Brazil., Brito CRDN; Departament of Clinical e Toxicological Analysis, Federal University of Rio Grande do Norte, Natal 59078-970, Brazil., Peres EG; Departament of Chemistry, Federal University of Amazonas, Manaus 69067-005, Brazil., de Lima Yamaguchi KK; Institute of Health and Biotecnology, Federal University of Amazonas, Coari 69460-000, Brazil., de Castro DP; Laboratory of Biochemistry and Physiology of Insects, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro 21040-360, Brazil., Moreira OC; Laboratory of Molecular Virology and Parasitology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro 21040-360, Brazil. |
| المصدر: | International journal of molecular sciences [Int J Mol Sci] 2024 May 18; Vol. 25 (10). Date of Electronic Publication: 2024 May 18. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Basel, Switzerland : MDPI, [2000- |
| مواضيع طبية MeSH: | Trypanosoma cruzi*/genetics , Trypanosoma cruzi*/isolation & purification , Chagas Disease*/epidemiology , Chagas Disease*/parasitology , Chagas Disease*/transmission , Chagas Disease*/diagnosis, Animals ; Real-Time Polymerase Chain Reaction/methods ; Euterpe ; Brazil/epidemiology ; Humans ; DNA, Protozoan/genetics |
| مستخلص: | Currently, approximately 70% of new cases of Chagas disease (CD) in Brazil are attributed to oral transmission, particularly through foods such as açaí, bacaba, and sugarcane juice, primarily in the northern and northeastern regions of the country. This underscores the imperative need to control the spread of the disease. The methods utilized to conduct quality control for food associated with outbreaks and to assess the potential for the oral transmission of CD through consuming açaí primarily rely on isolating the parasite or inoculating food into experimental animals, restricting the analyses to major research centers. While there are existing studies in the literature on the detection and quantification of T. cruzi DNA in açaí, the evaluation of parasites' viability using molecular methods in this type of sample and differentiating between live and dead parasites in açaí pulp remain challenging. Consequently, we developed a molecular methodology based on RT-qPCR for detecting and quantifying viable T. cruzi in açaí pulp samples. This protocol enables the stabilization and preservation of nucleic acids in açaí, along with incorporating an exogenous internal amplification control. The standardization of the RNA extraction method involved a simple and reproducible approach, coupled with a one-step RT-qPCR assay. The assay underwent validation with various T. cruzi DTUs and demonstrated sensitivity in detecting up to 0.1 viable parasite equivalents/mL in açaí samples. Furthermore, we investigated the effectiveness of a bleaching method in eliminating viable parasites in açaí samples contaminated with T. cruzi by comparing the detection of DNA versus RNA. Finally, we validated this methodology using açaí pulp samples positive for T. cruzi DNA, which were collected in a municipality with a history of oral CD outbreaks (Coari-AM). This validation involved comparing the detection and quantification of total versus viable T. cruzi . Collectively, our findings demonstrate the feasibility of this methodology in detecting viable forms of T. cruzi in açaí pulp samples, emerging as a crucial tool for monitoring oral outbreaks of Chagas disease resulting from açaí consumption. |
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| معلومات مُعتمدة: | 001 Coordenação de Aperfeicoamento de Pessoal de Nível Superior; 311539/2020-3 National Council for Scientific and Technological Development; CNE- E-26/201.096/2022 Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro; Inova Fiocruz/VPPCB/ VPPCB-007-FIO-18-2-72 Fundação Oswaldo Cruz |
| فهرسة مساهمة: | Keywords: Chagas disease; T. cruzi; açaí; one-step RT-qPCR; oral transmission; viability |
| المشرفين على المادة: | 0 (DNA, Protozoan) |
| تواريخ الأحداث: | Date Created: 20240525 Date Completed: 20240525 Latest Revision: 20240527 |
| رمز التحديث: | 20240527 |
| مُعرف محوري في PubMed: | PMC11122307 |
| DOI: | 10.3390/ijms25105531 |
| PMID: | 38791565 |
| قاعدة البيانات: | MEDLINE |
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