Acosta Soto Lucrecia, Santísima-Trinidad Ana Belén, Bornay-Llinares Fernando Jorge, Martín González Marcos, Pascual Valero José Antonio, Ros Muñoz Margarita
Department of Soil and Water Conservation and Organic Waste Management, Centro de Edafologia y Biologia Aplicada del Segura (CEBAS-CSIC), Campus de Espinardo, P.O. Box 164, Espinardo, 30100 Murcia, Spain; Área de Parasitología, Departamento de Agroquímica y Medio Ambiente, Universidad Miguel Hernández de Elche, Ctra Valencia Km 8.7, San Juan, 03550 Alicante, Spain.
Department of Soil and Water Conservation and Organic Waste Management, Centro de Edafologia y Biologia Aplicada del Segura (CEBAS-CSIC), Campus de Espinardo, P.O. Box 164, Espinardo, 30100 Murcia, Spain.
Biomed Res Int. 2017;2017:7515409. doi: 10.1155/2017/7515409. Epub 2017 Mar 9.
The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. is one of the soil-transmitted helminths (STH) with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO) recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR) and digital PCR (dPCR) were able to detect low amounts of eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one egg equivalent, while dPCR can detect DNA concentrations as low as five egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.
废水净化后再生水的回用在世界许多地区是一种广泛且必要的做法，并且必须伴随充分且持续的质量控制。蛔虫是土源性蠕虫之一，因其高传染性而对人类构成风险，而供水和卫生设施不足是传播的一个重要决定因素。世界卫生组织（WHO）建议，将再生水用于无限制灌溉时，每升水中寄生蠕虫卵的含量限制应等于或低于一个。我们提出了两种从大量再生水中提取DNA的新方案。定量PCR（qPCR）和数字PCR（dPCR）能够检测到少量的蛔虫卵。使用第一种处理500毫升再生水的提取方案时，qPCR能够检测到低至一个蛔虫卵当量的DNA浓度，而dPCR能够检测到低至五个蛔虫卵当量的DNA浓度。使用第二种处理10升再生水的方案时，qPCR能够检测到相当于20个蛔虫卵的DNA浓度。这一事实表明，开发新方法以更高的灵敏度和精度检测蠕虫卵、避免可能的人类感染风险非常重要。
