Quantitative detection of Neospora caninum in bovine aborted fetuses and experimentally infected mice by real-time PCR.

ABSTRACT: We report the development of a real-time PCR assay for the quantitative detection of Neospora caninum in infected host tissues. The assay uses the double-stranded DNA-binding dye SYBR Green I to continuously monitor product formation. Oligonucleotide primers were designed to amplify a 76-bp DNA fragment corresponding to the Nc5 sequence of N. caninum. A similar method was developed to quantify the 28S rRNA host gene in order to compare the parasite load of different samples and to correct for the presence of potential PCR-inhibiting compounds in the DNA samples. A linear quantitative detection range of 6 logs with a calculated detection limit of 10(-1) tachyzoite per assay was observed with excellent linearity (R(2) = 0.998). Assay specificity was confirmed by using DNA from the closely re

SUBMITTER: Collantes-Fernandez E 

PROVIDER: S-EPMC140374 | biostudies-literature | 2002 Apr

REPOSITORIES: biostudies-literature