ArticleApplied and environmental microbiology2001
Species-specific, nested PCR-restriction fragment length polymorphism detection of single Cryptosporidium parvum oocysts.
Article in Applied and environmental microbiology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed.
- Morphological and molecular characteristics of swine coccidia based on single oocyst isolation.Parasitology research · 2025Article
- Molecular characterisation and additional morphological descriptions of Eimeria spp. (Apicomplexa: Eimeriidae) from brown kiwi (Apteryx mantelli Bartlett).Systematic parasitology · 2023Article
- Potential Risk of Three Zoonotic Protozoa (Foods (Basel, Switzerland) · 2020Review
- Detection ofThe Korean journal of parasitology · 2018Article
- Cryptosporidium species and subtype analysis in diarrhoeic pre-weaned lambs and goat kids from north-western Spain.Parasitology research · 2015Article
- Distribution of indigenous bacterial pathogens and potential pathogens associated with roof-harvested rainwater.Applied and environmental microbiology · 2014Article
- Multicentric evaluation of a new real-time PCR assay for quantification of Cryptosporidium spp. and identification of Cryptosporidium parvum and Cryptosporidium hominis.Journal of clinical microbiology · 2013Article
- Article
- AIDS diarrhea and antiretroviral drug concentrations: a matched-pair cohort study in Port au Prince, Haiti.The American journal of tropical medicine and hygiene · 2011Article
- 90-kilodalton heat shock protein, Hsp90, as a target for genotyping Cryptosporidium spp. known to infect humans.Eukaryotic cell · 2009Article
- Detection of Cryptosporidium spp. from human faeces by PCR-RFLP, cloning and sequencing.Parasitology research · 2009Article
- Cryptosporidium species and subtypes and clinical manifestations in children, Peru.Emerging infectious diseases · 2008Article
- Epidemiology of cryptosporidiosis in North American travelers to Mexico.The American journal of tropical medicine and hygiene · 2008Article
- Evaluation of two DNA template preparation methods for post-immunomagnetic separation detection of Cryptosporidium parvum in foods and beverages by PCR.Applied and environmental microbiology · 2007Article
- Sensitivity of nested PCR in the detection of low numbers of Giardia lamblia cysts.Applied and environmental microbiology · 2007Article
- An evaluation of primers amplifying DNA targets for the detection of Cryptosporidium spp. using C. parvum HNJ-1 Japanese isolate in water samples.Parasitology research · 2007Article
- Detection and quantification of Cryptosporidium in HCT-8 cells and human fecal specimens using real-time polymerase chain reaction.The American journal of tropical medicine and hygiene · 2007Article
- Cryptosporidium taxonomy: recent advances and implications for public health.Clinical microbiology reviews · 2004Review
- Electrophoretic analysis of genetic variability within Cryptosporidium parvum from imported and autochthonous cases of human cryptosporidiosis in the United Kingdom.Applied and environmental microbiology · 2003Article
- Detection and differentiation of Cryptosporidium parasites that are pathogenic for humans by real-time PCR.Journal of clinical microbiology · 2002Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Concurrent with recent advances seen with Cryptosporidium parvum detection in both treated and untreated water is the need to properly evaluate these advances. A micromanipulation method by which known numbers of C. parvum oocysts, even a single oocyst, can be delivered to a test matrix for detection sensitivity is presented. Using newly developed nested PCR-restriction fragment length polymorphism primers, PCR sensitivity was evaluated with 1, 2, 3, 4, 5, 7, or 10 oocysts. PCR detection rates (50 samples for each number of oocysts) ranged from 38% for single oocysts to 92% for 5 oocysts, while 10 oocysts were needed to achieve 100% detection. The nested PCR conditions amplified products from C. parvum, Cryptosporidium baileyi, and Cryptosporidium serpentis but no other Cryptosporidium sp. or protozoan tested. Restriction enzyme digestion with VspI distinguished between C. parvum genotypes 1 and 2. Restriction enzyme digestion with DraII distinguished C. parvum from C. baileyi and C. serpentis. Use of known numbers of whole oocysts encompasses the difficulty of liberating DNA from the oocyst and eliminates the standard deviation inherent within a dilution series. To our knowledge this is the first report in which singly isolated C. parvum oocysts were used to evaluate PCR sensitivity. This achievement illustrates that PCR amplification of a single oocyst is feasible, yet sensitivity remains an issue, thereby illustrating the difficulty of dealing with low oocyst numbers when working with environmental water samples.
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