| Autor: |
Gobet, P, Buisson, J C, Vagner, O, Naciri, M, Grappin, M, Comparot, S, Harly, G, Aubert, D, Varga, I, Camerlynck, P, Bonnin, A |
| Zdroj: |
Journal of Clinical Microbiology; January 1997, Vol. 35 Issue: 1 p254-256, 3p |
| Abstrakt: |
We developed a PCR-based method that can be used to identify Cryptosporidium parvum in human feces. Fecal oocysts were concentrated by centrifugation on a sodium chloride gradient and filtration on a nitrocellulose filter prior to DNA extraction and PCR amplification of a 452-bp C. parvum-specific DNA sequence with a protocol including dUTP and uracil-N-glycosylase. All samples obtained from naturally infected humans (n = 10), calves (n = 4), and goats (n = 2) were positive. A 100% detection rate was achieved with both formed and solid stools (n = 10) seeded with 1,000 C. parvum oocysts per g. Procedures based on stool concentration by a modified Ritchie method and subsequent oocyst identification by immunofluorescent labeling or acid-fast staining require concentrations of 50,000 to 500,000 oocysts per g to achieve a 100% detection rate with formed stools. The described PCR-based assay thus has a 50- to 500-fold increase in sensitivity compared to those of the methods commonly used to analyze formed feces. |
| Databáze: |
Supplemental Index |
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