Ferreira Renata Trotta Barroso, Melandre Aline Martins, Cabral Maria Luiza, Branquinho Maria Regina, Cardarelli-Leite Paola
Programa de Pós-Graduação em Vigilância Sanitária, Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
Instituto Nacional de Controle de Qualidade em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, Rio de Janeiro, Brazil.
Rev Soc Bras Med Trop. 2016 Apr;49(2):190-5. doi: 10.1590/0037-8682-0414-2015.
Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met.
Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005.
DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important.
The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.
2004年以前，经口传播与食物相关的急性恰加斯病（ACD）的发生情况鲜为人知或鲜有研究。急性恰加斯病最初是散发性且偶发性的，现在在亚马逊地区已变得很常见，最近相关疫情已蔓延至巴西多个州。这些病例与流行地区被克氏锥虫感染的废物储存动物或昆虫媒介食用阿萨伊果汁有关。尽管存在将水果加工以尽量减少微生物和寄生虫污染的指南，但必须对阿萨伊基产品进行质量评估，为此必须满足对适当方法的需求。
将5至1000个克氏锥虫CL Brener细胞的稀释液与2毫升阿萨伊果汁混合。对该水果使用了四种克氏锥虫DNA提取方法，并根据欧盟联合研究中心（JRC）2005年的标准选择了十六烷基三甲基溴化铵（CTAB）法。
CTAB法提取的DNA在纯度和浓度方面均取得了令人满意的结果，可用于聚合酶链反应（PCR）。总体而言，所采用的方法证明，不仅提取效率，而且扩增中的高灵敏度都很重要。
食品中克氏锥虫的检测方法是疫情流行病学调查中的有力工具，因为它将流行病学证据转化为支持数据，用于确认食品中的克氏锥虫感染。它还有助于食品质量控制以及对涉及阿萨伊基产品的良好生产规范进行评估。
