title: Molecular detection of Toxoplasma gondii in water samples from Scotland and a comparison between the 529bp real-time PCR and ITS1 nested PCR.
abstract: Waterborne transmission of Toxoplasma gondii is a potential public health risk and there are currently no agreed optimised methods for the recovery, processing and detection of T. gondii oocysts in water samples. In this study modified methods of T. gondii oocyst recovery and DNA extraction were applied to 1427 samples collected from 147 public water supplies throughout Scotland. T. gondii DNA was detected, using real time PCR (qPCR) targeting the 529bp repeat element, in 8.79% of interpretable samples (124 out of 1411 samples). The samples which were positive for T. gondii DNA originated from a third of the sampled water sources. The samples which were positive by qPCR and some of the negative samples were reanalysed using ITS1 nested PCR (nPCR) and results compared. The 529bp qPCR was the more sensitive technique and a full analysis of assay performance, by Bayesian analysis using a Markov Chain Monte Carlo method, was completed which demonstrated the efficacy of this method for the detection of T. gondii in water samples.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: beth.wells@moredun.ac.uk.
authors.forename: Beth
authors.initials: B
authors.lastname: Wells
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: Hannah.shaw@moredun.ac.uk.
authors.forename: Hannah
authors.initials: H
authors.lastname: Shaw
authors.affiliation: Biomathematics and Statistics Scotland (BioSS), JCMB, King's Buildings, Peter Guthrie Tait Road, Edinburgh, EH9 3FD, Scotland, United Kingdom. Electronic address: giles@bioss.ac.uk.
authors.forename: Giles
authors.initials: G
authors.lastname: Innocent
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: Stefano.guido@moredun.ac.uk.
authors.forename: Stefano
authors.initials: S
authors.lastname: Guido
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: Emily.hotchkiss@moredun.ac.uk.
authors.forename: Emily
authors.initials: E
authors.lastname: Hotchkiss
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom; School of Agriculture and Veterinary Medicine, University of Bologna, Italy. Electronic address: maryparigi@gmail.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Parigi
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom; National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA, Bilthoven, The Netherlands. Electronic address: marieke.opsteegh@rivm.nl.
authors.forename: Marieke
authors.initials: M
authors.lastname: Opsteegh
authors.affiliation: Scottish Water, Juniper House, Heriot Watt Research Centre, Edinburgh, EH14 4AP, United Kingdom. Electronic address: J.Green@scottishwater.co.uk.
authors.forename: James
authors.initials: J
authors.lastname: Green
authors.affiliation: Scottish Water, Juniper House, Heriot Watt Research Centre, Edinburgh, EH14 4AP, United Kingdom. Electronic address: Simon.Gillespie@scottishwater.co.uk.
authors.forename: Simon
authors.initials: S
authors.lastname: Gillespie
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: lee.innes@moredun.ac.uk.
authors.forename: Elisabeth A
authors.initials: EA
authors.lastname: Innes
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Penicuik, Midlothian, EH26 0PZ, Scotland, United Kingdom. Electronic address: frank.katzer@moredun.ac.uk.
authors.forename: Frank
authors.initials: F
authors.lastname: Katzer
mesh_terms: D001499:Bayes Theorem; D016054:DNA, Protozoan / Q000235:genetics; D060766:Drinking Water / Q000469:parasitology*; D033661:Oocysts; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012606:Scotland; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014881:Water Supply
chemical_list: D016054:DNA, Protozoan; D060766:Drinking Water
keywords: Molecular detection; Public health; Toxoplasma gondii; Waterborne transmission