ArticleApplied and environmental microbiology1999
Detection of infectious Cryptosporidium parvum oocysts in surface and filter backwash water samples by immunomagnetic separation and integrated cell culture-PCR.
Article in Applied and environmental microbiology, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 142 citations in OpenAlex.
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- Public health significance of zoonoticInternational journal for parasitology. Parasites and wildlife · 2016Review
- Article
- Comparison of assays for sensitive and reproducible detection of cell culture-infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water.Applied and environmental microbiology · 2012Article
- Detection of viable Cryptosporidium parvum in soil by reverse transcription-real-time PCR targeting hsp70 mRNA.Applied and environmental microbiology · 2011Article
- Health risk from the use of roof-harvested rainwater in Southeast Queensland, Australia, as potable or nonpotable water, determined using quantitative microbial risk assessment.Applied and environmental microbiology · 2010Article
- Molecular characterization of sewage-borne pathogens and detection of sewage markers in an urban stream in Caracas, Venezuela.Applied and environmental microbiology · 2010Article
- Improved risk analysis by dual direct detection of total and infectious Cryptosporidium oocysts on cell culture in combination with immunofluorescence assay.Applied and environmental microbiology · 2010Article
- Cryptosporidium propidium monoazide-PCR, a molecular biology-based technique for genotyping of viable Cryptosporidium oocysts.Applied and environmental microbiology · 2009Article
- Application of PCR-based methods to assess the infectivity of enteric viruses in environmental samples.Applied and environmental microbiology · 2009Article
- Aged HCT-8 cell monolayers support Cryptosporidium parvum infection.Applied and environmental microbiology · 2007Article
- Detection of Toxoplasma gondii in water by an immunomagnetic separation method targeting the sporocysts.Parasitology research · 2007Article
- Detection of infectious Cryptosporidium oocysts by cell culture immunofluorescence assay: applicability to environmental samples.Applied and environmental microbiology · 2005Article
- Quantitative-PCR assessment of Cryptosporidium parvum cell culture infection.Applied and environmental microbiology · 2005Article
- Cryptosporidium taxonomy: recent advances and implications for public health.Clinical microbiology reviews · 2004Review
- Assessment of methods for detection of infectious Cryptosporidium oocysts and Giardia cysts in reclaimed effluents.Applied and environmental microbiology · 2003Article
- Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection.Applied and environmental microbiology · 2003Article
- Comparison of method 1623 and cell culture-PCR for detection of Cryptosporidium spp. in source waters.Applied and environmental microbiology · 2003Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A new strategy for the detection of infectious Cryptosporidium parvum oocysts in water samples, which combines immunomagnetic separation (IMS) for recovery of oocysts with in vitro cell culturing and PCR (CC-PCR), was field tested with a total of 122 raw source water samples and 121 filter backwash water grab samples obtained from 25 sites in the United States. In addition, samples were processed by Percoll-sucrose flotation and oocysts were detected by an immunofluorescence assay (IFA) as a baseline method. Samples of different water quality were seeded with viable C. parvum to evaluate oocyst recovery efficiencies and the performance of the CC-PCR protocol. Mean method oocyst recoveries, including concentration of seeded 10-liter samples, from raw water were 26.1% for IMS and 16.6% for flotation, while recoveries from seeded filter backwash water were 9.1 and 5.8%, respectively. There was full agreement between IFA oocyst counts of IMS-purified seeded samples and CC-PCR results. In natural samples, CC-PCR detected infectious C. parvum in 4.9% (6) of the raw water samples and 7.4% (9) of the filter backwash water samples, while IFA detected oocysts in 13.1% (16) of the raw water samples and 5.8% (7) of the filter backwash water samples. All CC-PCR products were confirmed by cloning and DNA sequence analysis and were greater than 98% homologous to the C. parvum KSU-1 hsp70 gene product. DNA sequence analysis also revealed reproducible nucleotide substitutions among the hsp70 fragments, suggesting that several different strains of infectious C. parvum were detected.
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