Evaluating Contamination Risk When Aspirating a Prosthetic Joint Using Microbial Next-Generation DNA Sequencing: An In Vitro Study.

Document Type

Article

Publication Date

12-16-2025

Publication Title

J Am Acad Orthop Surg Glob Res Rev

Abstract

BACKGROUND: A challenge in periprosthetic joint infection is the detection of microbiota. Next-generation sequencing (NGS) of microbial ribosomal DNA is one detection method, but false-positive results are of concern. We conducted a simulated joint aspiration study to assess false-positive rates with NGS testing.

METHODS: The simulated joint was a 50-mL bottle of sterile saline. Four techniques (n = 20 per technique) were tested. Fluid aspirates were inserted into sterile vacutainers with different needle exchanges analyzed for potential contamination points. The negative control group (n = 20 per technique) consisted of 80 sterile saline bottles tested directly at the NGS laboratory.

RESULTS: Eighty simulated aspirations were performed, two NGS tests per "joint." In three simulated techniques, the positive detection rate was 0.8%. In the fourth technique, where 10 mL of ambient air was aspirated into the syringe, the rate was 10%. The positive detection rate among the negative control bottles was 1.2%.

CONCLUSION: NGS detected DNA signals from sterile saline aspirations using a sterile technique. However, the false-positive rate was low (0.8%). We theorize that positive DNA signals originated from errant microbe contamination from ambient air drawn into the testing needle bore. We advocate needle exchange at every fluid transfer point.

Volume

9

Issue

12

DOI

10.5435/JAAOSGlobal-D-24-00144

ISSN

2474-7661

PubMed ID

41411511

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