On-farm colorimetric detection of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in crude bovine nasal samples

Vet Res. 2021 Oct 2;52(1):126. doi: 10.1186/s13567-021-00997-9.

Abstract

This work modifies a loop-mediated isothermal amplification (LAMP) assay to detect the bovine respiratory disease (BRD) bacterial pathogens Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in a colorimetric format on a farm. BRD causes a significant health and economic burden worldwide that partially stems from the challenges involved in determining the pathogens causing the disease. Methods such as polymerase chain reaction (PCR) have the potential to identify the causative pathogens but require lab equipment and extensive sample processing making the process lengthy and expensive. To combat this limitation, LAMP allows accurate pathogen detection in unprocessed samples by the naked eye allowing for potentially faster and more precise diagnostics on the farm. The assay developed here offers 66.7-100% analytical sensitivity, and 100% analytical specificity (using contrived samples) while providing 60-100% concordance with PCR results when tested on five steers in a feedlot. The use of a consumer-grade water bath enabled on-farm execution by collecting a nasal swab from cattle and provided a colorimetric result within 60 min. Such an assay holds the potential to provide rapid pen-side diagnostics to cattle producers and veterinarians.

Keywords: bovine respiratory disease; diagnostics; loop-mediated isothermal amplification; pen-side.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / microbiology
  • Colorimetry / instrumentation
  • Colorimetry / veterinary*
  • Diagnostic Tests, Routine / instrumentation
  • Diagnostic Tests, Routine / veterinary*
  • Mannheimia haemolytica / isolation & purification
  • Molecular Diagnostic Techniques / instrumentation
  • Molecular Diagnostic Techniques / veterinary*
  • Nose / microbiology
  • Nucleic Acid Amplification Techniques / instrumentation
  • Nucleic Acid Amplification Techniques / veterinary*
  • Pasteurella Infections / diagnosis
  • Pasteurella Infections / microbiology
  • Pasteurella Infections / veterinary
  • Pasteurella multocida / isolation & purification
  • Pasteurellaceae / isolation & purification*
  • Pasteurellaceae Infections / diagnosis
  • Pasteurellaceae Infections / microbiology
  • Pasteurellaceae Infections / veterinary*

Supplementary concepts

  • LAMP assay