Cryptosporidium and Giardia detection and discrimination of Cryptosporidium parvum genotypes by real-time PCR

摘要

We designed real-time PCR assays which amplify Cryptosporidium parvum sequences withmultiple nucleotide differences between type 1 and type 2. The presence of single-nucleotidepolymorphisms affected the amplicon melting temperature and the melting temperature ofinternal fluorescent probes, the basis for a type-specific diagnostic assay. By combining twopairs of PCR primers, one specific for Cryptosporidium parvum the other for Giardia lamblia, amultiplex assay for detecting both organisms was developed.