ArticleApplied and environmental microbiology2009
Cryptosporidium propidium monoazide-PCR, a molecular biology-based technique for genotyping of viable Cryptosporidium oocysts.
Article in Applied and environmental microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed.
- From conventional to biosensor-based detection ofFood and waterborne parasitology · 2026Review
- Refining microbial biomarker identification in rumen microbiome studies: a viability PCR-based approach.Applied and environmental microbiology · 2025Article
- Optimization ofJournal of environmental engineering (New York, N.Y.) · 2025Article
- Propidium monoazide PCR, a method to determine OsHV-1 undamaged capsids and to estimate virus Lethal Dose 50.Virus research · 2024Review
- Application of propidium monoazide quantitative PCR to discriminate of infectious African swine fever viruses.Frontiers in microbiology · 2023Article
- Cryptosporidiosis: From Prevention to Treatment, a Narrative Review.Microorganisms · 2022Review
- Propidium Monoazide Combined With RT-qPCR Detects Infectivity of Porcine Epidemic Diarrhea Virus.Frontiers in veterinary science · 2022Article
- Portable on-chip colorimetric biosensing platform integrated with a smartphone for label/PCR-free detection of Cryptosporidium RNA.Scientific reports · 2021Article
- A novel method for rapid and sensitive detection of viable Escherichia coli cells using UV-induced PMA-coupled quantitative PCR.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2020Article
- Review
- Evaluation of propidium monoazide-based qPCR to detect viable oocysts of Toxoplasma gondii.Parasitology research · 2019Article
- 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
- A comparison of methods to assess the antimicrobial activity of nanoparticle combinations on bacterial cells.PloS one · 2018Article
- Article
- Most probable number - loop mediated isothermal amplification (MPN-LAMP) for quantifying waterborne pathogens in <25min.Journal of microbiological methods · 2017Article
- Recent developments in detection and enumeration of waterborne bacteria: a retrospective minireview.MicrobiologyOpen · 2016Review
- 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
- Microbial succession in an inflated lunar/Mars analog habitat during a 30-day human occupation.Microbiome · 2016Article
- Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents.Journal of water and health · 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
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cryptosporidium is an important waterborne protozoan parasite that can cause severe diarrhea and death in the immunocompromised. The current methods used to monitor for Cryptosporidium oocysts in water are the microscopy-based USEPA methods 1622 and 1623. These methods assess total levels of oocysts in source waters, but do not determine oocyst viability or genotype. Recently, propidium monoazide (PMA) has been used in conjunction with molecular diagnostic tools to identify species and assess the viability of bacteria. The goal of this study was the development of a Cryptosporidium PMA-PCR (CryptoPMA-PCR) assay that includes PMA treatment prior to PCR analysis in order to prevent the amplification of DNA from dead oocysts. The results demonstrated that PMA penetrates only dead oocysts and blocks amplification of their DNA. The CryptoPMA-PCR assay can also specifically detect live oocysts within a mixed population of live and dead oocysts. More importantly, live oocysts, not dead oocysts, were detected in raw waste or surface water samples spiked with Cryptosporidium oocysts. This proof-of-concept study is the first to demonstrate the use of PMA for pre-PCR treatment of Cryptosporidium oocysts. The CryptoPMA-PCR assay is an attractive approach to specifically detect and genotype viable Cryptosporidium oocysts in the water, which is critical for human health risk assessment.
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