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Application of Nucleic Acid-Based Strategies to Detect Infectious Pathogens in Orthopaedic Implant-Related Infection
Emily Ann McClure1, Paul Werth2, Benjamin Ross1
1Dartmouth College, Hanover, New Hampshire.
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Implant-associated infection in orthopaedic surgery remains an enormous and largely unsolved clinical problem with a high rate of persistent or recurrent infection. This may be due, at least in part, to the potential for underdiagnosis by traditional microbial culture or the potential for culture to incompletely identify the microbial species present.
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Nucleic acid-based diagnostic techniques, focused on using the diagnostic information contained in DNA or RNA to identify microbial species, have been developing rapidly and have garnered escalating interest for both clinical and research applications.
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Commonly applied techniques include end-point polymerase chain reaction (PCR), quantitative PCR, Sanger sequencing, and next-generation sequencing. Understanding the specific strengths and weaknesses of each technique is critical to understanding their utility, applying the correct assessment strategy, and critically understanding and interpreting research.
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The best practices for interpreting nucleic acid-based diagnostic techniques include considering positive and negative controls, reads per sample, detection thresholds (for differentiating contaminants from positive results), and the primer set or targeted regions.
Insights
Orthopaedic implant infections are hard to diagnose with traditional methods. Nucleic acid-based diagnostics offer a promising alternative for accurate microbial identification, improving patient outcomes.
Area of Science:
- Orthopaedic Surgery
- Infectious Diseases
- Molecular Diagnostics
Background:
- Implant-associated infections in orthopaedic surgery present significant clinical challenges, often leading to persistent or recurrent infections.
- Traditional microbial culture methods may underdiagnose or incompletely identify causative microbial species, contributing to treatment failures.
Purpose of the Study:
- To review the rapidly developing field of nucleic acid-based diagnostic techniques for microbial identification in orthopaedic infections.
- To elucidate the strengths and weaknesses of commonly applied molecular methods.
- To outline best practices for interpreting results from these advanced diagnostic tools.
Main Methods:
- Review of nucleic acid-based diagnostic techniques including end-point polymerase chain reaction (PCR), quantitative PCR, Sanger sequencing, and next-generation sequencing.
- Discussion of critical factors for accurate interpretation of molecular diagnostic data.
Main Results:
- Nucleic acid-based techniques provide a powerful alternative to traditional culture for identifying microbial species.
- Understanding the specific characteristics of each molecular method is crucial for appropriate application and interpretation.
- Adherence to best practices in result interpretation enhances diagnostic accuracy.
Conclusions:
- Nucleic acid-based diagnostics are increasingly valuable for diagnosing orthopaedic implant-associated infections.
- Proper selection and interpretation of molecular techniques are essential for clinical and research success.
- Further research and standardized protocols will optimize the utility of these advanced diagnostic methods.
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