ArticleThe Korean journal of parasitology2014
DNA extraction from protozoan oocysts/cysts in feces for diagnostic PCR.
Article in The Korean journal of parasitology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers, 1 of them a synthesis that pooled it.
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Who cites it
30 citing papers in PubMed, 1 synthesis or guideline pooled it, 57 citations in OpenAlex.
- Gut protozoa of wild rodents - a meta-analysis.Parasitology · 2024Pooled it
- Molecular characterization and zoonotic potential ofFood and waterborne parasitology · 2026Article
- Genome-targeted enrichment and sequencing of human-infecting Cryptosporidium spp.Nature communications · 2026Article
- Development of optimized DNA extraction protocols for the quantification of Eimeria spp. from chicken feces using real-time PCR.Poultry science · 2026Article
- Pre-Analytical Cleanup of Feline Feces Improves DNA Extract Quality, Reduces Post-Extraction PCR Inhibition, and Enhances Molecular Detectability of Intestinal Protozoa.Current issues in molecular biology · 2026Article
- Genomic Characterization of Cryptosporidium spp. via iNextEra Library Preparation and Hybridization Bait Capture.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Evidence of Waterborne Parasites in Mussels for Human Consumption Harvested from a Recreational and Highly Productive Bay.Microorganisms · 2025Article
- Metagenomic detection of protozoan parasites on leafy greens aided by a rapid and efficient DNA extraction protocol.Frontiers in microbiology · 2025Article
- Article
- Enhancing diagnostic accuracy: Direct immunofluorescence assay as the gold standard for detecting Giardia duodenalis and Cryptosporidium spp. in canine and feline fecal samples.BMC veterinary research · 2024Article
- Nanoparticle Lysis ofMethods and protocols · 2024Article
- Multicenter comparative study of Enterocytozoon bieneusi DNA extraction methods from stool samples, and mechanical pretreatment protocols evaluation.Scientific reports · 2024Article
- Molecular Detection of a PathogenicPolish journal of microbiology · 2024Article
- An Epidemiological Assessment ofAnimals : an open access journal from MDPI · 2023Article
- Comparison Study of Four Extraction Methods Combined with PCR and LAMP for FelinePathogens (Basel, Switzerland) · 2022Article
- Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.Molecular biology reports · 2021Article
- Detection and Molecular Identification of Cryptosporidium Species Among Children with Malignancies.Acta parasitologica · 2021Article
- Prevalence and Phylogenetic Analysis of MicrosporidiumPathogens (Basel, Switzerland) · 2021Article
- Apparent lack of spill-over of parasites from an invasive anuran: PCR detectsInternational journal for parasitology. Parasites and wildlife · 2020Article
- Dientamoeba fragilis Infection in Patients with Digestive and Non-Digestive Symptoms: A Case-Control Study.The Korean journal of parasitology · 2020Article
Corrections and comments
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Authors and funding
1 author at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. QIAamp® DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume (50-100 µl) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, ≈ 2 oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.
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Registered trials
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