SYBR Green qPCR Technique for the Detection of Trypanosoma cruzi in Açaí Pulp. 2020

Gabrielle Virgínia Ferreira Cardoso, and Joelson Sousa Lima, and Andrey Carlos do Sacramento de Oliveira, and Josyane Brasil da Silva, and Talita Bandeira Roos, and Carina Martins de Moraes
Laboratory of Hygiene and Food Quality, Institute of Veterinary Medicine, Federal University of Pará (UFPA), Castanhal, Brazil.

Chagas disease, which is found widely in Latin America and has a great impact on public health, is caused by the parasite Trypanosoma cruzi. It is a neglected parasitic disease that urgently requires rapid diagnostic methods. The objective of this study was to develop a SYBR Green real-time quantitative polymerase chain reaction (qPCR) technique for the direct identification and quantification of T. cruzi from experimentally contaminated açai fruit samples. We used discrete typing units, TcI, containing 3.5 × 104 cells/mL, to infect the pulp of the açai fruit. This was followed by DNA extraction using a standardized procedure. The DNA samples were quantified and amplified at specific time and temperature intervals. The specificity of the oligoinitiators used in the qPCR assays was estimated by calculating the primer dissociation curve (melting curve) along with a detection threshold using different concentrations of DNA. The method used here demonstrated good efficiency and precision for the detection and quantification of T. cruzi DNA, with a detection limit of 2.65 × 10-14 g/μL DNA. The qPCR technique presented here could serve as an important tool for the diagnosis of T. cruzi parasites in açai.

UI MeSH Term Description Entries
D011804 Quinolines
D001938 Brazil A country located on the eastern coast of South America, located between Colombia and Peru, that borders the Atlantic Ocean. It is bordered on the north by Venezuela, Guyana, Suriname, and French Guiana, on the south by Uruguay, and on the west by Argentina. The capital is Brasilia.
D003959 Diamines Organic chemicals which have two amino groups in an aliphatic chain. Diamine
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000067030 Fruit and Vegetable Juices The liquids extracted from plants. Fruit Juices,Vegetable Juices,Fruit Juice,Juice, Fruit,Juice, Vegetable,Juices, Fruit,Juices, Vegetable,Vegetable Juice
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D014349 Trypanosoma cruzi The agent of South American trypanosomiasis or CHAGAS DISEASE. Its vertebrate hosts are man and various domestic and wild animals. Insects of several species are vectors. Trypanosoma cruzus,cruzi, Trypanosoma
D014355 Chagas Disease Infection with the protozoan parasite TRYPANOSOMA CRUZI, a form of TRYPANOSOMIASIS endemic in Central and South America. It is named after the Brazilian physician Carlos Chagas, who discovered the parasite. Infection by the parasite (positive serologic result only) is distinguished from the clinical manifestations that develop years later, such as destruction of PARASYMPATHETIC GANGLIA; CHAGAS CARDIOMYOPATHY; and dysfunction of the ESOPHAGUS or COLON. Trypanosomiasis, South American,American Trypanosomiasis,Chagas' Disease,Trypanosoma cruzi Infection,Infection, Trypanosoma cruzi,Infections, Trypanosoma cruzi,South American Trypanosomiasis,Trypanosoma cruzi Infections,Trypanosomiasis, American
D016054 DNA, Protozoan Deoxyribonucleic acid that makes up the genetic material of protozoa. Protozoan DNA

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