Pilot clinical trial: propidium monoazide PCR quantifies reduction of the viable bacterial load after antiseptic preparation of canine oral mucosa

Am J Vet Res. 2023 May 6;84(6):ajvr.23.02.0029. doi: 10.2460/ajvr.23.02.0029. Print 2023 Jun 1.

Abstract

Objective: A pilot clinical study to evaluate the use of propidium monoazide PCR (PMA-PCR) in quantifying a reduction of bacterial load after antiseptic use on the canine oral mucosa and skin, comparison of quantitative PCR (qPCR) to PMA-PCR, and comparison of patterns seen between PCR methods and bacterial culture.

Animals: Client-owned dogs (n = 10) undergoing general anesthesia and intravenous catheter placement.

Procedures: The oral mucosa and antebrachial skin of each dog underwent swabs for culture, qPCR, and PMA-PCR before and after antiseptic preparation of each site. Reduction in bacterial load between sampling times was evaluated for each quantification method.

Results: All testing methods found a significant decrease in bacterial load from oral mucosa after antiseptic preparation (culture P = .0020, qPCR P = .0039, PMA-PCR P = .0039). PMA-PCR had a significantly greater reduction of bacterial load after preparation than qPCR (P = .0494). Only culture detected a significant reduction after preparation of the skin (culture P = .0039, qPCR P = .3125, PMA-PCR P = .0703).

Clinical relevance: PMA-PCR was able to quantify a reduction of bacterial load after antiseptic preparation of the high-bacterial load environment, with a pattern similar to that of culture, and was more specific than qPCR for detecting viable bacterial load. The results of this study support the use of PMA-PCR for antiseptic effectiveness studies performed on a high-bacterial load environment, such as canine oral mucosa.

MeSH terms

  • Animals
  • Anti-Infective Agents, Local* / pharmacology
  • Anti-Infective Agents, Local* / therapeutic use
  • Azides / pharmacology
  • Bacterial Load / methods
  • Bacterial Load / veterinary
  • Dogs
  • Mouth Mucosa
  • Propidium
  • Real-Time Polymerase Chain Reaction / veterinary

Substances

  • propidium monoazide
  • Anti-Infective Agents, Local
  • Propidium
  • Azides