Author: Elia, Gabriella; Cavalli, Alessandra; Desario, Costantina; Lorusso, Eleonora; Lucente, Maria Stella; Decaro, Nicola; Martella, Vito; Buonavoglia, Canio
Title: Detection of infectious canine parvovirus type 2 by mRNA real-time RT-PCR Cord-id: 6notzm6s Document date: 2007_8_10
ID: 6notzm6s
Snippet: A TaqMan real-time RT-PCR assay was developed for detection of RNA transcripts produced by replicating CPV-2. A pair of primers and a TaqMan probe targeting the spliced NS2 mRNA were designed. A synthetic DNA fragment was constructed to mimic the spliced NS2 mRNA by PCR-based gene assembly and was used for generation of standard RNAs. The detection limit of the assay was 1 × 10(2) RNA copies and standard curve displayed a linear range from 1 × 10(2) to 1 × 10(9) copies and a good reproducibil
Document: A TaqMan real-time RT-PCR assay was developed for detection of RNA transcripts produced by replicating CPV-2. A pair of primers and a TaqMan probe targeting the spliced NS2 mRNA were designed. A synthetic DNA fragment was constructed to mimic the spliced NS2 mRNA by PCR-based gene assembly and was used for generation of standard RNAs. The detection limit of the assay was 1 × 10(2) RNA copies and standard curve displayed a linear range from 1 × 10(2) to 1 × 10(9) copies and a good reproducibility. The assay was then applied to determine the mRNA loads in the tissues of dogs naturally infected by CPV-2. mRNA was detected in a variety of tissues, including the central nervous system.
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