Development of a TaqMan quantitative PCR assay specific for Cryptosporidium parvum.
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.
ONDEO Services, Centre Technique et de Recherche, Paris, 38, Avenue du President Wilson, 78230 Le Pecq, France.
Melanie
M
Fontaine
Emmanuelle
E
Guillot
D000818:Animals; D016785:Cryptosporidium parvum / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D016054:DNA, Protozoan / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D019914:Taq Polymerase / Q000378:metabolism*; D014867:Water / Q000469:parasitology
D016054:DNA, Protozoan; D014867:Water; D019914:Taq Polymerase