Multiplex PCR-based next generation sequencing as a novel, targeted and accurate molecular approach for periprosthetic joint infection diagnosis

Changyu Huang; Ying Huang; Ziwen Wang; Yiming Lin; Yongbin Li; Yang Chen; Xiaoqing Chen; Chaofan Zhang; Wenbo Li; Wenming Zhang; Xinyu Fang; Zida Huang

doi:10.3389/fmicb.2023.1181348

Multiplex PCR-based next generation sequencing as a novel, targeted and accurate molecular approach for periprosthetic joint infection diagnosis

Front Microbiol. 2023 May 18:14:1181348. doi: 10.3389/fmicb.2023.1181348. eCollection 2023.

Authors

Changyu Huang^{1

2

3}, Ying Huang^{1

2

3}, Ziwen Wang⁴, Yiming Lin¹, Yongbin Li¹, Yang Chen¹, Xiaoqing Chen⁵, Chaofan Zhang^{1

2

3}, Wenbo Li^{1

2

3}, Wenming Zhang^{1

2

3}, Xinyu Fang^{1

2

3}, Zida Huang^{1

2

3}

Affiliations

¹ Department of Orthopaedic Surgery, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
² Department of Orthopedic Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
³ Fujian Provincial Institute of Orthopedics, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
⁴ Fujian Medical University, Fuzhou, China.
⁵ Department of Orthopedic Surgery, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, China.

Abstract

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.

Keywords: culture; diagnostics; metagenomic next-generation sequencing; periprosthetic joint infection; targeted next-generation sequencing.