Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1997 Jan;35(1):254-6.
doi: 10.1128/jcm.35.1.254-256.1997.

Detection of Cryptosporidium parvum DNA in formed human feces by a sensitive PCR-based assay including uracil-N-glycosylase inactivation

P Gobet  1 , J C Buisson, O Vagner, M Naciri, M Grappin, S Comparot, G Harly, D Aubert, I Varga, P Camerlynck, A Bonnin
Affiliations

Detection of Cryptosporidium parvum DNA in formed human feces by a sensitive PCR-based assay including uracil-N-glycosylase inactivation

P Gobet et al. J Clin Microbiol. 1997 Jan.

Abstract

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.

PubMed Disclaimer

References

    1. J Infect Dis. 1986 Feb;153(2):365-8 - PubMed
    1. J Infect Dis. 1995 Oct;172(4):1160-4 - PubMed
    1. J Infect Dis. 1988 Feb;157(2):225-9 - PubMed
    1. Crit Rev Microbiol. 1988;16(2):113-59 - PubMed
    1. Ann Intern Med. 1990 Jan 1;112(1):75-6 - PubMed

Publication types