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Evaluating Contamination Risk When Aspirating a Prosthetic Joint Using Microbial Next-Generation DNA Sequencing: An
Edward J McPherson1, Madhav Chowdhry, Devin R Young
1From the Departmenrt of Orthopaedic Surgery, David Geffen School of Medicine, UCLA, Los Angeles, CA (Dr. McPherson and Dr. Stavrakis); the Nuffield Department of Primary Care Health Sciences and Department of Continuing Education, Kellogg College, University of Oxford, Oxford, United Kingdom (Dr. Chowdhry); and the Oakland University William Beaumont School of Medicine, Rochester, MI (Mr. Young).
Next-generation sequencing (NGS) for detecting microbes in joint infections has a low false-positive rate of 0.8% when sterile techniques are used. Contamination from ambient air is a likely source of false positives.
Area of Science:
- Microbiology
- Genomics
- Medical Diagnostics
Background:
- Detecting microbiota in periprosthetic joint infections is challenging.
- Next-generation sequencing (NGS) of microbial ribosomal DNA is a potential diagnostic method.
- Concerns exist regarding false-positive results with NGS testing.
Purpose of the Study:
- To assess the false-positive rates of NGS testing in simulated joint aspirations.
- To identify potential sources of contamination during the aspiration process.
Main Methods:
- A simulated joint aspiration study was conducted using sterile saline in 50-mL bottles.
- Four different aspiration techniques were tested, with 20 replicates per technique.
- Fluid aspirates were transferred to sterile vacutainers, analyzing contamination points.
- A negative control group of 80 sterile saline bottles was tested directly at the NGS laboratory.
Main Results:
- The overall positive detection rate across three simulated techniques was 0.8%.
- The fourth technique, involving aspiration of 10 mL of ambient air, showed a 10% positive detection rate.
- The positive detection rate in the negative control bottles was 1.2%.
Conclusions:
- NGS can detect DNA signals from sterile saline aspirations using sterile techniques.
- The study found a low false-positive rate of 0.8% with proper sterile technique.
- Errant microbe contamination from ambient air drawn into the needle bore is theorized as a source of false positives.
- Advocating for needle exchange at every fluid transfer point is recommended to minimize contamination.
