ArticleJournal of clinical microbiology2013
Multicentric evaluation of a new real-time PCR assay for quantification of Cryptosporidium spp. and identification of Cryptosporidium parvum and Cryptosporidium hominis.
Article in Journal of clinical microbiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
40 citing papers in PubMed, 83 citations in OpenAlex.
- 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
- Molecular characterization and phylogenetic analysis of cryptosporidium spp. in pediatric acute gastroenteritis: Epidemiological insights from northeastern Iran.New microbes and new infections · 2025Article
- Comparative analysis of commercial and "In-House" molecular tests for the detection of intestinal protozoa in stool samples.Parasites & vectors · 2025Article
- [High prevalence ofMedecine tropicale et sante internationale · 2024Article
- Is Personal Protective Equipment Worth the Hassle? Annual Risk of Cryptosporidiosis to Dairy Farmers and How Personal Protective Equipment and Handwashing Can Mitigate It.Microorganisms · 2023Article
- Evaluation of the Performance of the NovodiagPathogens (Basel, Switzerland) · 2023Article
- Rapid Reverse Purification DNA Extraction Approaches to Identify Microbial Pathogens in Wastewater.Microorganisms · 2023Article
- Dual Transcriptomics To Determine Gamma Interferon-Independent Host Response to Intestinal Cryptosporidium parvum Infection.Infection and immunity · 2022Article
- Cryptosporidiosis outbreak in Amazonia, French Guiana, 2018.PLoS neglected tropical diseases · 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
- PersistentMicroorganisms · 2021Article
- Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis ofPathogens (Basel, Switzerland) · 2021Article
- Comparative Performance of Eight PCR Methods to DetectPathogens (Basel, Switzerland) · 2021Article
- Comparative Study of Eleven Mechanical Pretreatment Protocols forMicroorganisms · 2021Article
- DisseminatedIDCases · 2021Article
- Multicenter Comparative Study of SixMicroorganisms · 2020Article
- DetectingThe American journal of tropical medicine and hygiene · 2020Article
- Article
- Asymptomatic Cryptosporidium infections in ewes and lambs are a source of environmental contamination with zoonotic genotypes of Cryptosporidium parvum.Parasite (Paris, France) · 2020Article
- Assessment of differences between DNA content of cell-cultured and freely suspended oocysts of Cryptosporidium parvum and their suitability as DNA standards in qPCR.Parasites & vectors · 2019Article
Corrections and comments
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Authors and funding
13 authors at 9 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cryptosporidium is a protozoan parasite responsible for gastroenteritis, especially in immunocompromised patients. Laboratory diagnosis of cryptosporidiosis relies on microscopy, antigen detection, and nucleic acid detection and analysis. Among the numerous molecular targets available, the 18S rRNA gene displays the best sensitivity and sequence variations between species and can be used for molecular typing assays. This paper presents a new real-time PCR assay for the detection and quantification of all Cryptosporidium species associated with the identification of Cryptosporidium hominis and Cryptosporidium parvum. The sensitivity and specificity of this new PCR assay were assessed on a multicentric basis, using well-characterized Cryptosporidium-positive and -negative human stool samples, and the efficiencies of nine extraction methods were comparatively assessed using Cryptosporidium-seeded stool samples and phosphate-buffered saline samples. A comparison of extraction yields showed that the most efficient extraction method was the Boom technique in association with mechanical grinding, and column extraction showed higher binding capacity than extraction methods based on magnetic silica. Our PCR assay was able to quantify at least 300 oocysts per gram of stool. Satisfactory reproducibility between laboratories was observed. The two main species causing human disease, Cryptosporidium hominis and Cryptosporidium parvum, were identified using a duplex real-time PCR assay with specific TaqMan minor-groove-binding ligand (MGB) probes for the same amplicon. To conclude, this one-step quantitative PCR is well suited to the routine diagnosis of cryptosporidiosis since practical conditions, including DNA extraction, quantification using well-defined standards, and identification of the two main species infecting humans, have been positively assessed.
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