title: Selective detection of viable Cryptosporidium oocysts by PCR.
abstract: Although the diagnosis of cryptosporidiosis in diarrheic patients is relatively simple, as large amounts of oocysts are usually shed, environmental samples can contain only few oocysts which have a comparatively high epidemiological relevance. Very sensitive detection methods are therefore requested in environmental hygiene. Additionally these methods should allow a statement about the viability of the detected organisms. The combination of a DNA-digest and a viability assay followed by a PCR might reveal a very promising method. A DNA-digest destroys free DNA in the sample. In the following excystation protocol only viable sporozoites excyst. A PCR carried out now gives definitive proof of viable oocysts. Dead sporozoites within intact oocyst-walls are not detected.
authors.affiliation: Abteilung für Allgemeine Hygiene und Umwelthygiene, Eberhard-Karls-Universität Tübingen.
authors.forename: R
authors.initials: R
authors.lastname: Filkorn
authors.forename: A
authors.initials: A
authors.lastname: Wiedenmann
authors.forename: K
authors.initials: K
authors.lastname: Botzenhart
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003458:Cryptosporidium / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis* / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan