Development of a TaqMan quantitative PCR assay specific forCryptosporidium parvum
- Resource Type
- Authors
- Emmanuelle Guillot; Melanie Fontaine
- Source
- FEMS Microbiology Letters. 214:13-17
- Subject
- Serial dilution
animal diseases
Polymerase Chain Reaction
Sensitivity and Specificity
Microbiology
DNA sequencing
law.invention
Species Specificity
law
parasitic diseases
Genetics
TaqMan
Animals
Taq Polymerase
Molecular Biology
Polymerase chain reaction
Cryptosporidium parvum
biology
Water
DNA, Protozoan
biology.organism_classification
Virology
Molecular biology
Orders of magnitude (mass)
genomic DNA
Real-time polymerase chain reaction
- Language
- ISSN
- 1574-6968
0378-1097
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.