title: Comparison of diagnostic techniques for the detection of Cryptosporidium oocysts in animal samples.
abstract: While a large number of laboratory methods for the detection of Cryptosporidium oocysts in faecal samples are now available, their efficacy for identifying asymptomatic cases of cryptosporidiosis is poorly understood. This study was carried out to determine a reliable screening test for epidemiological studies in livestock. In addition, three molecular tests were compared to identify Cryptosporidium species responsible for the infection in cattle, sheep and horses. A variety of diagnostic tests including microscopic (Kinyoun's staining), immunological (Direct Fluorescence Antibody tests or DFAT), enzyme-linked immunosorbent assay (ELISA), and molecular methods (nested PCR) were compared to assess their ability to detect Cryptosporidium in cattle, horse and sheep faecal samples. The results indicate that the sensitivity and specificity of each test is highly dependent on the input samples; while Kinyoun's and DFAT proved to be reliable screening tools for cattle samples, DFAT and PCR analysis (targeted at the 18S rRNA gene fragment) were more sensitive for screening sheep and horse samples. Finally different PCR primer sets targetedat the same region resulted in the preferential amplification of certain Cryptosporidium species when multiple species were present in the sample. Therefore, for identification of Cryptosporidium spp. in the event of asymptomatic cryptosporidiosis, the combination of different 18S rRNA nested PCR primer sets is recommended for further epidemiological applications and also tracking the sources of infection.
authors: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; School of Public Health and Population Science, University College Dublin, Belfield, Dublin 4, Ireland. Electronic address: marzieh.mirhashemi@tufts.edu.
authors: Marzieh
authors: M
authors: Ezzaty Mirhashemi
authors: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.
authors: Annetta
authors: A
authors: Zintl
authors: School of Public Health and Population Science, University College Dublin, Belfield, Dublin 4, Ireland.
authors: Tim
authors: T
authors: Grant
authors: Centre for Environmental Research Innovation and Sustainability (CERIS), Institute of Technology, Sligo, Ireland.
authors: Frances E
authors: FE
authors: Lucy
authors: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.
authors: Grace
authors: G
authors: Mulcahy
authors: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.
authors: Theo
authors: T
authors: De Waal
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D003457:Cryptosporidiosis / Q000175:diagnosis*; D003458:Cryptosporidium / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005243:Feces / Q000469:parasitology; D019085:Fluorescent Antibody Technique, Direct / Q000662:veterinary; D006734:Horse Diseases / Q000175:diagnosis* / Q000469:parasitology; D006736:Horses; D008403:Mass Screening / Q000379:methods / Q000662:veterinary; D033661:Oocysts; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000469:parasitology; D013194:Staining and Labeling / Q000379:methods / Q000662:veterinary
chemical_list: D016054:DNA, Protozoan; D012337:RNA, Ribosomal, 18S
keywords: Cattle; Cryptosporidium; Diagnostic techniques; Horse; Latent class analysis; Molecular methods; Sheep