ArticleApplied and environmental microbiology2005
Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates and for relief of PCR inhibitors.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 83 papers.
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Who cites it
83 citing papers in PubMed, 238 citations in OpenAlex.
- Occurrence and Genetic Diversity ofAnimals : an open access journal from MDPI · 2026Article
- Mouse Models for Use in Cryptosporidium Infection Studies, Quantification of Parasite Burden Using Flow Cytometry, qPCR and Histopathology, and Confocal Imaging of Oocysts.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Animal Species and Identity Testing: Developments, Challenges, and Applications to Non-Human Forensics.Genes · 2025Review
- PCR optimization for buccal swab-derived samples: overcoming sporadic inhibition with bovine serum albumin.BMC genomics · 2025Article
- Monitoring of Adenoviruses in Water and Wastewater through Applications of PCR Based Methods.Indian journal of microbiology · 2025Review
- First molecular identification of Cryptosporidium species isolated from canal water bodies in Minya Al-Qamh district, Northern Egypt.BMC research notes · 2025Article
- Occurrence and Molecular Characterization ofMicroorganisms · 2025Article
- Identification of two new genetic loci for high-resolution genotyping of Enterocytozoon bieneusi.Parasite (Paris, France) · 2025Article
- Distribution of human-pathogenicFrontiers in microbiology · 2025Article
- High prevalence and pathogenicity ofCurrent research in parasitology & vector-borne diseases · 2025Article
- Prevalence ofInternational journal of veterinary science and medicine · 2025Article
- Genetic characterization of Cryptosporidium spp. in the North African hedgehog (Atelerix algirus) in the Canary Islands, Spain.Parasitology research · 2024Article
- Assessment of the potential occurrence ofJournal of parasitic diseases : official organ of the Indian Society for Parasitology · 2024Article
- Molecular identification and antiprotozoal activity of silver nanoparticles on viability of Cryptosporidium parvum isolated from pigeons, pigeon fanciers and water.Scientific reports · 2023Article
- High infectivity and unique genomic sequence characteristics of Cryptosporidium parvum in China.PLoS neglected tropical diseases · 2022Article
- Detection ofMicroorganisms · 2022Review
- High zoonotic potential of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in wild nonhuman primates from Yunnan Province, China.Parasites & vectors · 2022Article
- DivergentFrontiers in cellular and infection microbiology · 2022Article
- Standardization of molecular techniques for the detection and characterization of intestinal protozoa and other pathogens in humans.The journal of venomous animals and toxins including tropical diseases · 2022Article
- Padlock probe-based rolling circle amplification lateral flow assay for point-of-need nucleic acid detection.The Analyst · 2021Article
23 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Extraction of high-quality DNA is a key step in PCR detection of Cryptosporidium and other pathogens in environmental samples. Currently, Cryptosporidium oocysts in water samples have to be purified from water concentrates before DNA is extracted. This study compared the effectiveness of six DNA extraction methods (DNA extraction with the QIAamp DNA minikit after oocyst purification with immunomagnetic separation and direct DNA extraction methods using the FastDNA SPIN kit for soil, QIAamp DNA stool minikit, UltraClean soil kit, or QIAamp DNA minikit and the traditional phenol-chloroform technique) for the detection of Cryptosporidium with oocyst-seeded samples, DNA-spiked samples, and field water samples. The study also evaluated the effects of different PCR facilitators (nonacetylated bovine serum albumin, the T4 gene 32 protein, and polyvinylpyrrolidone) and treatments (the use of GeneReleaser or ultrafiltration) for the relief from or removal of inhibitors of PCR amplification. The results of seeding and spiking studies showed that PCR inhibitors were presented in all DNA solutions extracted by the six methods. However, the effect of PCR inhibitors could be relieved significantly by the addition of 400 ng of bovine serum albumin/mul or 25 ng of T4 gene 32 protein/mul to the PCR mixture. With the inclusion of bovine serum albumin in the PCR mixture, DNA extracted with the FastDNA SPIN kit for soil without oocyst isolation resulted in PCR performance similar to that produced by the QIAamp DNA minikit after oocysts were purified by immunomagnetic separation.
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