Laboratory of Hygiene and Food Quality, Institute of Veterinary Medicine, Federal University of Pará (UFPA), Castanhal, Brazil.
Undergraduate Course in Nutrition Maurício de Nassau University (UniNassau), Belém, Brazil.
Foodborne Pathog Dis. 2020 Jul;17(7):466-469. doi: 10.1089/fpd.2019.2745. Epub 2020 Feb 20.
Chagas disease, which is found widely in Latin America and has a great impact on public health, is caused by the parasite . 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 from experimentally contaminated açai fruit samples. We used discrete typing units, TcI, containing 3.5 × 10 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 DNA, with a detection limit of 2.65 × 10 g/μL DNA. The qPCR technique presented here could serve as an important tool for the diagnosis of parasites in açai.
恰加斯病广泛存在于拉丁美洲，对公共卫生有重大影响，是由寄生虫引起的。它是一种被忽视的寄生虫病，迫切需要快速诊断方法。本研究旨在开发一种 SYBR Green 实时定量聚合酶链反应（qPCR）技术，用于直接鉴定和定量从实验污染的巴西莓果样本中的寄生虫。我们使用离散分型单位 TcI，含有 3.5×10 个细胞/毫升，感染巴西莓果肉。随后采用标准化程序提取 DNA。在特定的时间和温度间隔内对 DNA 样本进行定量和扩增。通过使用不同浓度的 DNA 计算引物解离曲线（熔解曲线）和检测阈值，评估 qPCR 检测中使用的寡核苷酸启动子的特异性。这里使用的方法显示了检测和定量 DNA 寄生虫的良好效率和精度，检测限为 2.65×10 g/μL DNA。本文提出的 qPCR 技术可作为巴西莓中寄生虫诊断的重要工具。
