Multiplex real-time PCR in the detection and differentiation of bovine respiratory disease pathogens

Bovine respiratory syncytial virus (BRSV), bovine viral diarrhea (BVD) and bovine parainfluenza virus Type 3 (BPI3) are ubiquitous respiratory pathogens of cattle, which contribute to causation of bovine respiratory disease complex. As these respiratory viral pathogens cause very similar clinical symptoms, differential diagnosis of the pathogens is required in one sample. Therefore, a single tube fluorogenic multiplex real time-PCR-based TaqMan assay was developed for simultaneous detection of BRSV and BPI3 from bovine respiratory samples. TaqMan-PCR was optimized to quantify the viruses using the BioRad iCycler IQ real-time PCR detection system and dual-labeled fluorogenic probes. Primers and probes for our study were designed from a 100% conserved region of BRSV N gene and from BPI3 NP gene using GenBank sequences such that they can be used to differentiate BRSV and BPI3 in one-tube in clinical respiratory samples obtained from bovine species. Once the technique was optimized with validated reference strains and field isolates, the assay was applied to routine diagnostic samples. Respiratory samples collected from 100 cattle across the state of Oregon were screened for BRSV and BPI3 viruses by real time-PCR and virus isolation. The monoplex real-time PCR identified 19 samples positive for BRSV, 11 for BPI-3 and 14 for BVD. The multiplex real-time PCR identified 18 samples positive for BRSV and 4 samples positive for BPI3 Among positive samples, 2 samples showed positive for both BRSV and BPI3. BVD was left out of the multiplex reaction due to primer interference. In conclusion, the multiplex real-time TaqMan-PCR described here for detection and quantitation of BRSV and BPI3 viruses has been shown to be sensitive and specific. Rapid turnaround time, reproducibility and ease of use make this technique a valuable diagnostic tool for detection of BRSV and BPI3 in individual or pooled respiratory samples

The Vehicle, 1967, Vol. 10 no. 1

Vol. 10, No. 1 Table of Contents Christy Bowercover Photographpage 4 The CityCatherine Waitepage 4 A New DealAnonymouspage 5 The Penny Ride of Carnation PinkAstaire Pappaspage 8 RapeSharon Nelsonpage 10 Born AgainCharles J. Mertzpage 10 DrawingRourkepage 11 UntitledRourkepage 11 DialogueMolly Evanspage 12 SeldomJamse Jonsepage 12 The DifferenceDennis Muchmorepage 13 First LoveBruce Czeluscinskipage 15 Photographpage 15 Immorality of Troilus and Criseyde Milo S. Metcalfpage 16 Answer UnknownDave Owenpage 20 I Am LostJean Lacypage 21 Photographpage 22 On the Eve of No TomorrowsJane Careypage 23 Unrelated TwinsByron Nelsonpage 24 if i sitRoger Zulaufpage 25 if i sitDennis Muchmorepage 26 Drawingpage 27 Bibliographypage 28https://thekeep.eiu.edu/vehicle/1015/thumbnail.jp

The Vehicle, 1969, Vol. 11 no. 2

Vol. 11, No. 2 Table of Contents Short Story: The TripCharles Whitepage 4 PhotoDale Huberpage 5 A NightRoger Zulaufpage 6 Sixteen-year-old Students...NBpage 6 LostJim Biropage 6 The AmateurRoger Zulaufpage 7 ManRoger Zulaufpage 7 My CamelotRonald Garnerpage 7 The Rose and the BriarKenneth L. Folkertspage 9 Who Am I?Frank McKennedypage 10 PhotoDale Huberpage 11 Mr. Samuel ClemensLarry A. Millerpage 11 Lock OutAra Childspage 12 Excuse MeRoger Zulaufpage 12 On Shadows from a Candle \u2767Michael G. McKeepage 12 beginning of an endCaryl Dagropage 12 DrawingMADpage 13 We Ain\u27t Un HurJames Birchlerpage 13 Genesis II, 18 \u2767Michael G. McKeepage 13 Short Story: A Patent Leather PaleEleanor Aikenpage 14 hungry childRoger Zulaufpage 15 DrawingRoger Zulaufpage 15 PhotoRoger Digglepage 16 Do You Like The Rain?Linda Boltmanpage 17 Seasons ChangePerry J. Carterpage 17 PhotoDale Huberpage 19 Whistling TreesPam McKinneypage 19 PostscriptThomas W. Reapage 20 PhotoDale Huberpage 20https://thekeep.eiu.edu/vehicle/1020/thumbnail.jp