Comparison of a mycobacterial phage assay to detect viable Mycobacterium avium subspecies paratuberculosis with standard diagnostic modalities in cattle with naturally infected Johne disease.

Robert J. Greenstein*, Liya Su, Irene R. Grant, Antonio Foddai, Amy Turner, Jason S Nagati, Sheldon T. Brown, Judith R. Stabel

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Downloads (Pure)

Abstract

Background Mycobacterium avium subspecies paratuberculosis (MAP), the cause of Johne disease, is a slow growing mycobacterium. Viable MAP detection is difficult, inconstant and time-consuming. The purpose of this study was to compare a rapid phage/qPCR assay performed on peripheral blood mononuclear cells (PBMCs) with three standard methods of MAP detection: fecal MAP PCR; plasma antigen-specific IFN-γ & serum MAP ELISA hypothesizing that, if sensitive and specific, Johne animals would be positive and Control animals negative. We studied a well characterized herd of Holstein cattle that were naturally infected with MAP and their Controls. Results With phage/qPCR 72% (23/32) of Johne and 35% (6/17) of Controls were MAP positive. With fecal PCR 75% (24/32) of Johne and 0% (0/17) of Controls were MAP positive. With plasma antigen-specific IFN-γ 69% (22/32) of Johne and 12% (2/17) of Controls were MAP positive. With serum MAP ELISA, 31% (10/32) of Johne and 0% (0/17) of Controls were MAP positive. When phage / qPCR and fecal PCR results were combined, 100% (32/32) Johne and 35% (6/17) of Control animals were MAP positive. Younger Control animals (1–3 years) had significantly fewer plaques (25 ± 17 SEM) than older Controls (4–12 years) (309 ± 134 p = 0.04). The same trend was not observed in the Johne animals (p = 0.19). Conclusions In contrast to our hypothesis, using the phage/qPCR assay we find that viable circulating MAP can rapidly be detected from the blood of animals infected with, as well as those in the Control group evidently colonized by MAP. These data indicate that the presence of viable MAP in blood does not necessarily signify that an animal must of necessity be demonstrably ill or be MAP positive by standard diagnostic methods.
Original languageEnglish
Article number30
Number of pages12
JournalGut Pathogens
Volume13
DOIs
Publication statusPublished - 06 May 2021

Keywords

  • Mycobacterium avium subsp. paratuberculosis (MAP)
  • mycobacteriophage D29
  • Johne's disease
  • veterinary diagnostics
  • mycobacteria

Fingerprint Dive into the research topics of 'Comparison of a mycobacterial phage assay to detect viable<i> Mycobacterium avium</i> subspecies <i>paratuberculosis </i>with standard diagnostic modalities in cattle with naturally infected Johne disease.'. Together they form a unique fingerprint.

Cite this