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BioMed Research International
Volume 2017, Article ID 7515409, 9 pages
https://doi.org/10.1155/2017/7515409
Research Article

Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

1Department 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
2Área de Parasitología, Departamento de Agroquímica y Medio Ambiente, Universidad Miguel Hernández de Elche, Ctra Valencia Km 8.7, San Juan, 03550 Alicante, Spain
3Aguas de Murcia, C/Rincón de San Antón, Llano de Brujas, 30161 Murcia, Spain

Correspondence should be addressed to Lucrecia Acosta Soto; se.hmu@atsocal

Received 28 November 2016; Revised 10 January 2017; Accepted 16 January 2017; Published 9 March 2017

Academic Editor: Pratik Banerjee

Copyright © 2017 Lucrecia Acosta Soto et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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. Ascaris lumbricoides 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 A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.