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Applied and Environmental Microbiology, October 1999, p. 4431-4435, Vol. 65, No. 10
Division of Parasitic Diseases, National
Center for Infectious Diseases, Centers for Disease Control and
Prevention, Atlanta, Georgia 30341
Received 21 December 1998/Accepted 10 July 1999
We evaluated the specificity and sensitivity of 11 previously
described species differentiation and genotyping PCR protocols for
detection of Cryptosporidium parasites. Genomic DNA from
three species of Cryptosporidium parasites (genotype 1 and
genotype 2 of C. parvum, C. muris, and C. serpentis), two Eimeria species (E. neischulzi and E. papillata), and Giardia
duodenalis were used to evaluate the specificity of primers.
Furthermore, the sensitivity of the genotyping primers was tested by
using genomic DNA isolated from known numbers of oocysts obtained from
a genotype 2 C. parvum isolate. PCR amplification was
repeated at least three times with all of the primer pairs. Of the 11 protocols studied, 10 amplified C. parvum genotypes 1 and
2, and the expected fragment sizes were obtained. Our results indicate
that two species-differentiating protocols are not
Cryptosporidium specific, as the primers used in these
protocols also amplified the DNA of Eimeria species. The
sensitivity studies revealed that two nested PCR-restriction fragment
length polymorphism (RFLP) protocols based on the small-subunit rRNA
and dihydrofolate reductase genes are more sensitive than single-round
PCR or PCR-RFLP protocols.
0099-2240/99/$04.00+0
Evaluation of Cryptosporidium parvum
Genotyping Techniques
*
Corresponding author. Mailing address: Division of
Parasitic Diseases, National Center for Infectious Diseases, Centers
for Disease Control and Prevention, Building 22, Mail Stop F-12, 4770 Buford Highway, Atlanta, GA 30341-3717. Phone: (770) 488-4047. Fax:
(770) 488-4454. E-mail: AAL1{at}CDC.GOV.
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