ArticleJournal of clinical microbiology2005
Detection of cryptosporidium and identification to the species level by nested PCR and restriction fragment length polymorphism.
Article in Journal of clinical microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 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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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
31 citing papers in PubMed, 118 citations in OpenAlex.
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- Source tracking of fecal contamination in Asian green mussels (Journal of parasitic diseases : official organ of the Indian Society for Parasitology · 2024Article
- An Epidemiological Assessment ofAnimals : an open access journal from MDPI · 2023Article
- Article
- Efficient nested-PCR-based method development for detection and genotype identification of Acanthamoeba from a small volume of aquatic environmental sample.Scientific reports · 2021Article
- Challenges forGenes · 2021Review
- Performance of three rapid diagnostic tests for the detection of Cryptosporidium spp. and Giardia duodenalis in children with severe acute malnutrition and diarrhoea.Infectious diseases of poverty · 2019Article
- Cryptosporidium spp., prevalence, molecular characterisation and socio-demographic risk factors among immigrants in Qatar.PLoS neglected tropical diseases · 2019Article
- Detection and molecular characterization of Cryptosporidium species and Giardia assemblages in two watersheds in the metropolitan region of São Paulo, Brazil.Environmental science and pollution research international · 2018Article
- First detection of Cryptosporidium DNA in blood and cerebrospinal fluid of HIV-infected patients.Parasitology research · 2018Article
- Microscopic and Molecular Tracing of Cryptosporidium Oocysts: Identifying a Possible Reservoir of Infection in Red Grouse.Pathogens (Basel, Switzerland) · 2017Article
- Molecular Analysis of the Enteric Protozoa Associated with Acute Diarrhea in Hospitalized Children.Frontiers in cellular and infection microbiology · 2017Article
- Occurrence of Giardia intestinalis and Cryptosporidium sp. in wastewater samples from São Paulo State, Brazil, and Lima, Peru.Environmental science and pollution research international · 2016Article
- Molecular Identification of Cryptosporidium Species from Pet Snakes in Thailand.The Korean journal of parasitology · 2016Article
- INTESTINAL AND PULMONARY INFECTION BY Cryptosporidium parvum IN TWO PATIENTS WITH HIV/AIDS.Revista do Instituto de Medicina Tropical de Sao Paulo · 2016Article
- Genotyping of Cryptosporidium Species and Their Clinical Manifestations in Patients with Renal Transplantation and Human Immunodeficiency Virus Infection.Journal of pathogens · 2016Article
- Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina.Memorias do Instituto Oswaldo Cruz · 2016Article
- Molecular and parasitological study of cryptosporidium isolates from cattle in ilam, west of iran.Iranian journal of parasitology · 2014Article
- Molecular Characterization of Cryptosporidium spp. Isolated From Immunocompromised Patients and Children.Jundishapur journal of microbiology · 2014Article
Corrections and comments
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cryptosporidiosis is an emerging protozoan disease associated with large waterborne outbreaks. Diagnosis relies on microscopic examination of stools, but this method cannot identify the infecting species of Cryptosporidium. We have developed a test based on nested PCR and restriction fragment length polymorphism (RFLP) that offers simple identification of Cryptosporidium hominis, Cryptosporidium parvum, and most other human infective species in stool samples. Purified C. parvum oocysts were used for PCR development. Extracted DNA was amplified by nested PCR targeting a 214-bp fragment of the 18S RNA gene. Enzymatic restriction sites were identified by bioinformatic analysis of all published Cryptosporidium 18S rRNA sequences. Experiments with spiked stool samples gave an estimated PCR detection limit of one oocyst. Specificity was assessed by testing 68 stool samples from patients with microscopically proven cryptosporidiosis and 31 Cryptosporidium-negative stools. Sixty-seven (98.5%) of the 68 stool samples from patients with microscopically proven cryptosporidiosis and 2 of the other stool samples were positive by PCR and could be genotyped. RFLP analysis identified 36 C. hominis, 19 C. parvum, 8 Cryptosporidium meleagridis, and 6 Cryptosporidium felis or Cryptosporidium canis samples. Species determination in 26 PCR-positive cases was in full agreement with DNA sequencing of the 18S rRNA hypervariable region. The excellent sensitivity of PCR, coupled with the accuracy of RFLP for species identification, make this method a suitable tool for routine diagnosis and genotyping of Cryptosporidium in stools.
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