Bibliographic Details
| Title: |
A novel multiplex qPCR targeting 23S rDNA for diagnosis of swine dysentery and porcine intestinal spirochaetosis. |
| Authors: |
Borgström, Anna1 anna.borgstroem@vetbakt.uzh.ch, Scherrer, Simone1, Kirchgässner, Constanze1, Schmitt, Sarah1, Frei, Daniel1, Wittenbrink, Max M.1 |
| Source: |
BMC Veterinary Research. 2/7/2017, p1-8. 8p. |
| Subject Terms: |
*SWINE disease diagnosis, *DYSENTERY, *TREPONEMA hyodysenteriae, *SPIROCHAETOSIS, *POLYMERASE chain reaction, *SURGICAL swabs, *DIAGNOSIS |
| Abstract: |
Background: A multiplex qPCR targeting a 128 bp region on the 23S rDNA gene was developed for detection of Brachyspira (B.) hyodysenteriae and B. pilosicoli, the agents of swine dysentery (SD) and porcine intestinal spirochaetosis (PIS), together with a triplet of apathogenic Brachyspira spp. (B. innocens, B. intermedia, B. murdochii) in porcine feces. The multiplex qPCR was evaluated against a duplex PCR (La et al., J Clin Microbiol 41:3372-5, 2003). Results: Using DNA extracted from fecal culture, the multiplex qPCR showed excellent agreement with the duplex PCR (k = 0.943 and 0.933). In addition, thanks to the three probes whereof one detecting the apathogenic Brachyspria spp., a more diversified overview of the brachyspiral flora in porcine fecal samples can be delivered as a part of the routine diagnostic. The multiplex qPCR with a limit of detection of 5-10 genomic equivalents (GE) per reaction (6 × 102 GE per gram) allows reliable detection of Brachyspira species directly from fecal swab DNA. In line with this, analysis of 202 fecal swabs in comparison with culture-based qPCR showed a high agreement for the causative agents of SD (B.hyodysenteriae: k = 0.853, sensitivity 87% specificity 98%). Conclusion: The novel multiplex qPCR is robust and has a high analytical sensitivity and is therefore suitable for high-throughput screening of porcine fecal swabs for the causative agents of SD. This assay can therefore be used for the direct proof of the pathogenic B. spp. in fecal swabs within the scope of a monitoring program. [ABSTRACT FROM AUTHOR] |
|
Copyright of BMC Veterinary Research is the property of BioMed Central and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) |
| Database: |
Academic Search Complete |
