Related Experiment Video
Updated: Jul 27, 2025
Novel Diagnostics in Revision Arthroplasty: Implant Sonication and Multiplex Polymerase Chain Reaction
Published on: December 3, 2017
Multiplex PCR-based next generation sequencing as a novel, targeted and accurate molecular approach for
Changyu Huang1,2,3, Ying Huang1,2,3, Ziwen Wang4
1Department of Orthopaedic Surgery, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
Objectives:
Periprosthetic joint infection (PJI) diagnosis remains challenging, and the identification of the causative microorganism is, by far, the most important aspect. Here, we use multiple PCR-based targeted next-generation sequencing (tNGS) to detect pathogens in PJI. To explore 1. the ability of targeted next-generation sequencing (tNGS) to detect pathogens in PJI; 2. the consistency of tNGS, metagenomic NGS (mNGS), and culture results; and 3. the ability of tNGS to detect drug resistance genes in PJI.
Methods:
PJI was diagnosed according to the Musculoskeletal Infection Society (MSIS) criteria. The microorganisms were detected by culture, mNGS and tNGS to compare the diagnostic effectiveness of the three methods for PJI and to compare their consistency in detecting microorganisms. Drug resistance genes were detected using tNGS. The costs and turnaround times of mNGS and tNGS were compared.
Results:
Forty-three patients with PJI, 21 patients without PJI and 10 negative control cases were included. The culture, tNGS, and mNGS sensitivities for PJI diagnosis were 74.41%, 88.37%, and 93.02%, respectively, with no significant differences. The specificities were 90.48%, 95.24%, and 95.24%, respectively, with no significant differences. tNGS detected drug resistance genes in 37.5% of culture-positive PJIs. tNGS was superior to mNGS for turnaround time (14.5 h vs. 28 h) and cost ($150 vs. $260).
Conclusions:
tNGS can effectively identify PJI pathogens and may provide drug resistance information, while tNGS is superior to mNGS regarding cost and turnaround time. A multidisciplinary, multi-technology based algorithm to diagnose PJI is appropriate.
Highlights:
298 microorganisms and 86 drug resistance genes were included in the tNGS panel.Diagnostic efficacy of tNGS is not inferior to that of commonly used indicators.tNGS is superior to mNGS in cost and turnaround time.
Insights
Targeted next-generation sequencing (tNGS) effectively identifies pathogens in periprosthetic joint infection (PJI) and detects drug resistance genes. This method offers faster turnaround times and lower costs compared to metagenomic sequencing, making it a valuable diagnostic tool.
Area of Science:
- Microbiology
- Genomics
- Infectious Diseases
Background:
- Periprosthetic joint infection (PJI) diagnosis is challenging, with pathogen identification being critical.
- Current diagnostic methods for PJI have limitations in speed and comprehensive pathogen detection.
Purpose of the Study:
- To evaluate the efficacy of targeted next-generation sequencing (tNGS) in detecting pathogens in PJI.
- To compare the consistency of tNGS with metagenomic NGS (mNGS) and traditional culture methods.
- To assess the ability of tNGS to identify antimicrobial resistance genes in PJI pathogens.
Main Methods:
- Patients with and without PJI, along with negative controls, were analyzed.
- Microorganisms were detected using culture, mNGS, and tNGS.
- Diagnostic sensitivities and specificities of the methods were compared.
- tNGS was used to detect drug resistance genes, and cost-effectiveness was evaluated.
Main Results:
- tNGS demonstrated high sensitivity (88.37%) and specificity (95.24%) for PJI diagnosis, comparable to mNGS and culture.
- tNGS identified drug resistance genes in 37.5% of culture-positive PJIs.
- tNGS offered a significantly shorter turnaround time (14.5 hours vs. 28 hours) and lower cost ($150 vs. $260) than mNGS.
Conclusions:
- tNGS is an effective tool for identifying PJI pathogens and can provide crucial drug resistance information.
- tNGS presents advantages over mNGS in terms of cost and turnaround time for PJI diagnostics.
- A multidisciplinary approach integrating multiple technologies is recommended for optimal PJI diagnosis.
