Detection and Genotyping of Oocysts of Cryptosporidium parvum by Real-Time PCR and Melting Curve Analysis

摘要: Several real-time PCR procedures for the detection and genotyping of oocysts Cryptosporidium parvum were evaluated. A 40-cycle amplification a 157-bp fragment from C. β-tubulin gene detected individual which introduced into reaction mixture by micromanipulation. SYBR Green I melting curve analysis was used to confirm specificity method when DNA extracted fecal samples spiked with analyzed. Because isolates infecting humans comprise two distinct genotypes, designated type 1 2, methods discriminating genotypes developed. The first same primers fluorescently labeled antisense oligonucleotide probes spanning 49-bp polymorphic sequence diagnostic 2. second fluorescence targeted coding region within GP900/poly(T) gene. Both discriminated between 2 on basis analysis. To our knowledge, this is report describing application oocysts.