Open AccessDevelopment of a TaqMan quantitative PCR assay specific for Cryptosporidium parvum
Author(s) -
Fontaine Melanie,
Guillot Emmanuelle
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11318.x
Subject(s) - cryptosporidium parvum , taqman , serial dilution , primer (cosmetics) , biology , polymerase chain reaction , real time polymerase chain reaction , microbiology and biotechnology , cryptosporidium , genomic dna , apicomplexa , dna , chemistry , protozoal disease , gene , genetics , feces , medicine , alternative medicine , organic chemistry , pathology , malaria , immunology
A rapid detection method that is both quantitative and specific for the water‐borne human parasite Cryptosporidium parvum is reported. Real‐time polymerase chain reaction (PCR) combined with fluorescent TaqMan technology was used to develop this sensitive and accurate assay. The selected primer–probe set identified a 138‐bp section specific to a C. parvum genomic DNA sequence. The method was optimized on a cloned section of the target DNA sequence, then evaluated on C. parvum oocyst dilutions. Quantification was accomplished by comparing the fluorescence signals obtained from test samples of C. parvum oocysts with those obtained from standard dilutions of C. parvum oocysts. This real‐time PCR assay allowed reliable quantification of C. parvum oocysts over six orders of magnitude with a baseline sensitivity of six oocysts in 2 h.