ArticleParasitology research2014
Quantification of viable Giardia cysts and Cryptosporidium oocysts in wastewater using propidium monoazide quantitative real-time PCR.
Article in Parasitology research, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Climate-Sensitive Redistribution of Veterinary Parasites: An Attribution Framework for One Health Surveillance and Control.Biology · 2026Review
- Optimization ofJournal of environmental engineering (New York, N.Y.) · 2025Article
- Molecular Identification of ProtozoanLife (Basel, Switzerland) · 2022Article
- Hastening Progress inMicroorganisms · 2022Article
- Detection ofMicroorganisms · 2022Review
- Optimization and validation of a loop-mediated isothermal amplification (LAMP) assay for detection ofFood and waterborne parasitology · 2021Article
- Determination of the viability of Toxoplasma gondii oocysts by PCR real-time after treatment with propidium monoazide.Revista do Instituto de Medicina Tropical de Sao Paulo · 2020Article
- Evaluation of propidium monoazide-based qPCR to detect viable oocysts of Toxoplasma gondii.Parasitology research · 2019Article
- The occurrence ofGastroenterology and hepatology from bed to bench · 2019Article
- Detection and molecular characterization of Cryptosporidium species and Giardia assemblages in two watersheds in the metropolitan region of São Paulo, Brazil.Environmental science and pollution research international · 2018Article
- Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts) of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods.Parasite (Paris, France) · 2018Review
- Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania: genetic characterization and public health potential of the isolates.Environmental science and pollution research international · 2017Article
- Quantitative detection of viable helminth ova from raw wastewater, human feces, and environmental soil samples using novel PMA-qPCR methods.Environmental science and pollution research international · 2016Article
- Detection and Quantification of Viable and Nonviable Trypanosoma cruzi Parasites by a Propidium Monoazide Real-Time Polymerase Chain Reaction Assay.The American journal of tropical medicine and hygiene · 2016Article
- Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents.Journal of water and health · 2016Article
- Advances and Challenges in Viability Detection of Foodborne Pathogens.Frontiers in microbiology · 2016Review
- Development and Evaluation of Three Real-Time PCR Assays for Genotyping and Source Tracking Cryptosporidium spp. in Water.Applied and environmental microbiology · 2015Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Real-time PCR (qPCR) is a rapid tool to quantify pathogens in the aquatic environment; however, it quantifies all pathogens, including both viable and nonviable. Propidium monoazide (PMA) is a membrane-impairment dye that penetrates only membrane-damaged cells. Once inside the cell, PMA is covalently cross-linked to DNA through light photoactivation, and PCR amplification is strongly inhibited. The goal of this study was to evaluate PMA-qPCR assays for rapid quantification of viable and heat-treated Giardia cysts and Cryptosporidium oocysts in wastewater. We observed a reduction in detection of heat-treated Giardia duodenalis cysts of 83.2, 89.9, 98.2, or 97% with PMA-qPCR assays amplifying a 75 base-pair (bp) β-giardin target, 77-bp triosephosphate isomerase (tpi), 133-bp glutamate dehydrogenase (GDH), and 143-bp β-giardin gene target, respectively. Thus, the exclusion of dead cysts was more effective when qPCR assays that produced larger amplicons were used. The PMA treatment of Cryptosporidium oocysts plus/minus heat treatment abolished the fluorescent signal for dead oocysts with a PMA-qPCR assay amplifying a Cryptosporidium parvum (150-bp) oocyst wall protein (COWP) gene. The PMA-qPCR 143-bp β-giardin assay for Giardia and the PMA-qPCR 150-bp COWP assay for Cryptosporidium accurately quantified live oo(cysts), and failed to detect dead oo(cysts), when phosphate-buffered saline and tertiary effluent wastewater were spiked with concentrations of 10(3) or 10(2) dead oo(cysts), respectively. Therefore, these assays are suitable for the detection of viable parasites that are typically present in tertiary wastewater effluents at concentrations of <10(3) oo(cysts)/l and can provide rapid risk assessments of environmental water.
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