Diagnostic accuracy of 16S rDNA PCR, multiplex PCR and metagenomic next-generation sequencing in periprosthetic joint infections: a systematic review and meta-analysis

The diagnostic accuracy of 16S rDNA PCR, multiplex PCR (mPCR), and metagenomic next-generation sequencing (mNGS) in periprosthetic joint infections (PJIs) remains unclear. This study aims to evaluate the diagnostic accuracy of 16S rDNA PCR, mPCR, and mNGS in PJI. Data sources: Data sources included...

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Published in:	Clinical microbiology and infection Vol. 31; no. 7; p. 1115
Main Authors:	Olearo, Flaminia, Zein, Said El, Portillo, Maria Eugenia, Zapf, Antonia, Rohde, Holger, Berbari, Elie F, Wouthuyzen-Bakker, Marjan
Format:	Journal Article
Language:	English
Published:	England 01.07.2025
Subjects:	DNA, Ribosomal - genetics High-Throughput Nucleotide Sequencing - methods Humans Metagenomics - methods Molecular Diagnostic Techniques - methods Multiplex Polymerase Chain Reaction - methods Polymerase Chain Reaction - methods Prosthesis-Related Infections - diagnosis Prosthesis-Related Infections - microbiology RNA, Ribosomal, 16S - genetics Sensitivity and Specificity 16S rDNA PCR Multiplex PCR Metagenomic next-generation sequencing 16S rDNA Diagnostic accuracy tests Prosthetic joint infection
ISSN:	1469-0691, 1469-0691
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Abstract	The diagnostic accuracy of 16S rDNA PCR, multiplex PCR (mPCR), and metagenomic next-generation sequencing (mNGS) in periprosthetic joint infections (PJIs) remains unclear. This study aims to evaluate the diagnostic accuracy of 16S rDNA PCR, mPCR, and mNGS in PJI. Data sources: Data sources included PubMed and Embase (January 1, 2000-March 1, 2024), with no language restrictions. Studies containing sufficient data to construct a 2 × 2 contingency table allowing for sensitivity and specificity calculation were considered. Participants included adults (≥18 years) with PJI and appropriate control groups. Tests included 16S rDNA PCR, mPCR, and mNGS. Diagnosis required adherence to Musculoskeletal Infection Society, Infectious Diseases Society of America, International Consensus Meeting, European Bone and Joint Infection Society criteria. Studies employing alternative author-defined criteria were included only if they did not rely solely on positive cultures to define PJI. Quality Assessment of Diagnostic Accuracy Studies 2 was used. A bivariate model calculated pooled diagnostic odds ratios (DORs), sensitivities, and specificities, each with 95% CIs. Seventy-nine studies were included, comprising 3940 PJI cases and 4700 uninfected controls. Pooled sensitivity/specificity were 80.0% (95% CI, 75.4-84.3%)/94.0% (95% CI, 91-96%) for 16S rDNA PCR; 62.2% (52.5-70.9%)/96.2% (93.2-97.9%) for mPCR; and 88.6% (83.3-92.4%)/93.2% (89.5-95.6%) for mNGS. Notably, mNGS had the highest DOR (105.9; 95% CI, 60-186.9). A sensitivity analysis excluding lower-quality studies resulted in increased DORs for all methods. These molecular techniques display strong diagnostic accuracy for identifying PJI. Although mNGS yielded the highest DOR, numerous technical and practical challenges preclude its routine use for PJI diagnosis. Significant heterogeneity across studies warrants cautious interpretation and underscores the need for future comparative research.
AbstractList	The diagnostic accuracy of 16S rDNA PCR, multiplex PCR (mPCR), and metagenomic next-generation sequencing (mNGS) in periprosthetic joint infections (PJIs) remains unclear.BACKGROUNDThe diagnostic accuracy of 16S rDNA PCR, multiplex PCR (mPCR), and metagenomic next-generation sequencing (mNGS) in periprosthetic joint infections (PJIs) remains unclear.This study aims to evaluate the diagnostic accuracy of 16S rDNA PCR, mPCR, and mNGS in PJI.OBJECTIVESThis study aims to evaluate the diagnostic accuracy of 16S rDNA PCR, mPCR, and mNGS in PJI.Data sources: Data sources included PubMed and Embase (January 1, 2000-March 1, 2024), with no language restrictions.METHODSData sources: Data sources included PubMed and Embase (January 1, 2000-March 1, 2024), with no language restrictions.Studies containing sufficient data to construct a 2 × 2 contingency table allowing for sensitivity and specificity calculation were considered.STUDY ELIGIBILITY CRITERIAStudies containing sufficient data to construct a 2 × 2 contingency table allowing for sensitivity and specificity calculation were considered.Participants included adults (≥18 years) with PJI and appropriate control groups.PARTICIPANTSParticipants included adults (≥18 years) with PJI and appropriate control groups.Tests included 16S rDNA PCR, mPCR, and mNGS.TESTSTests included 16S rDNA PCR, mPCR, and mNGS.Diagnosis required adherence to Musculoskeletal Infection Society, Infectious Diseases Society of America, International Consensus Meeting, European Bone and Joint Infection Society criteria. Studies employing alternative author-defined criteria were included only if they did not rely solely on positive cultures to define PJI.REFERENCE STANDARDDiagnosis required adherence to Musculoskeletal Infection Society, Infectious Diseases Society of America, International Consensus Meeting, European Bone and Joint Infection Society criteria. Studies employing alternative author-defined criteria were included only if they did not rely solely on positive cultures to define PJI.Quality Assessment of Diagnostic Accuracy Studies 2 was used.ASSESSMENT OF RISK OF BIASQuality Assessment of Diagnostic Accuracy Studies 2 was used.A bivariate model calculated pooled diagnostic odds ratios (DORs), sensitivities, and specificities, each with 95% CIs.METHODS OF DATA SYNTHESISA bivariate model calculated pooled diagnostic odds ratios (DORs), sensitivities, and specificities, each with 95% CIs.Seventy-nine studies were included, comprising 3940 PJI cases and 4700 uninfected controls. Pooled sensitivity/specificity were 80.0% (95% CI, 75.4-84.3%)/94.0% (95% CI, 91-96%) for 16S rDNA PCR; 62.2% (52.5-70.9%)/96.2% (93.2-97.9%) for mPCR; and 88.6% (83.3-92.4%)/93.2% (89.5-95.6%) for mNGS. Notably, mNGS had the highest DOR (105.9; 95% CI, 60-186.9). A sensitivity analysis excluding lower-quality studies resulted in increased DORs for all methods.RESULTSSeventy-nine studies were included, comprising 3940 PJI cases and 4700 uninfected controls. Pooled sensitivity/specificity were 80.0% (95% CI, 75.4-84.3%)/94.0% (95% CI, 91-96%) for 16S rDNA PCR; 62.2% (52.5-70.9%)/96.2% (93.2-97.9%) for mPCR; and 88.6% (83.3-92.4%)/93.2% (89.5-95.6%) for mNGS. Notably, mNGS had the highest DOR (105.9; 95% CI, 60-186.9). A sensitivity analysis excluding lower-quality studies resulted in increased DORs for all methods.These molecular techniques display strong diagnostic accuracy for identifying PJI. Although mNGS yielded the highest DOR, numerous technical and practical challenges preclude its routine use for PJI diagnosis. Significant heterogeneity across studies warrants cautious interpretation and underscores the need for future comparative research.DISCUSSIONThese molecular techniques display strong diagnostic accuracy for identifying PJI. Although mNGS yielded the highest DOR, numerous technical and practical challenges preclude its routine use for PJI diagnosis. Significant heterogeneity across studies warrants cautious interpretation and underscores the need for future comparative research. The diagnostic accuracy of 16S rDNA PCR, multiplex PCR (mPCR), and metagenomic next-generation sequencing (mNGS) in periprosthetic joint infections (PJIs) remains unclear. This study aims to evaluate the diagnostic accuracy of 16S rDNA PCR, mPCR, and mNGS in PJI. Data sources: Data sources included PubMed and Embase (January 1, 2000-March 1, 2024), with no language restrictions. Studies containing sufficient data to construct a 2 × 2 contingency table allowing for sensitivity and specificity calculation were considered. Participants included adults (≥18 years) with PJI and appropriate control groups. Tests included 16S rDNA PCR, mPCR, and mNGS. Diagnosis required adherence to Musculoskeletal Infection Society, Infectious Diseases Society of America, International Consensus Meeting, European Bone and Joint Infection Society criteria. Studies employing alternative author-defined criteria were included only if they did not rely solely on positive cultures to define PJI. Quality Assessment of Diagnostic Accuracy Studies 2 was used. A bivariate model calculated pooled diagnostic odds ratios (DORs), sensitivities, and specificities, each with 95% CIs. Seventy-nine studies were included, comprising 3940 PJI cases and 4700 uninfected controls. Pooled sensitivity/specificity were 80.0% (95% CI, 75.4-84.3%)/94.0% (95% CI, 91-96%) for 16S rDNA PCR; 62.2% (52.5-70.9%)/96.2% (93.2-97.9%) for mPCR; and 88.6% (83.3-92.4%)/93.2% (89.5-95.6%) for mNGS. Notably, mNGS had the highest DOR (105.9; 95% CI, 60-186.9). A sensitivity analysis excluding lower-quality studies resulted in increased DORs for all methods. These molecular techniques display strong diagnostic accuracy for identifying PJI. Although mNGS yielded the highest DOR, numerous technical and practical challenges preclude its routine use for PJI diagnosis. Significant heterogeneity across studies warrants cautious interpretation and underscores the need for future comparative research.
Author	Rohde, Holger Portillo, Maria Eugenia Berbari, Elie F Zein, Said El Zapf, Antonia Wouthuyzen-Bakker, Marjan Olearo, Flaminia
Author_xml	– sequence: 1 givenname: Flaminia surname: Olearo fullname: Olearo, Flaminia organization: Department of Microbiology, Institute for Medical Microbiology, Virology and Hygiene, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany – sequence: 2 givenname: Said El surname: Zein fullname: Zein, Said El organization: Division of Public Health, Infectious Diseases and Occupational Medicine, Department of Medicine, Mayo Clinic, Rochester, MN, USA – sequence: 3 givenname: Maria Eugenia surname: Portillo fullname: Portillo, Maria Eugenia organization: Department of Clinical Microbiology, University Hospital of Navarra, Instituto de Investigación Sanitaria de Navarra-CIBERESP, Pamplona, Spain – sequence: 4 givenname: Antonia surname: Zapf fullname: Zapf, Antonia organization: Department of Microbiology, Institute of Medical Biometry and Epidemiology, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany – sequence: 5 givenname: Holger surname: Rohde fullname: Rohde, Holger organization: Department of Microbiology, Institute for Medical Microbiology, Virology and Hygiene, Universitätsklinikum Hamburg-Eppendorf, Hamburg, Germany – sequence: 6 givenname: Elie F surname: Berbari fullname: Berbari, Elie F organization: Division of Public Health, Infectious Diseases and Occupational Medicine, Department of Medicine, Mayo Clinic, Rochester, MN, USA – sequence: 7 givenname: Marjan surname: Wouthuyzen-Bakker fullname: Wouthuyzen-Bakker, Marjan email: m.wouthuyzen-bakker@umcg.nl organization: Department of Medical Microbiology and Infection Prevention, The University Medical Center Groningen, The University of Groningen, Groningen, The Netherlands. Electronic address: m.wouthuyzen-bakker@umcg.nl
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SubjectTerms	DNA, Ribosomal - genetics High-Throughput Nucleotide Sequencing - methods Humans Metagenomics - methods Molecular Diagnostic Techniques - methods Multiplex Polymerase Chain Reaction - methods Polymerase Chain Reaction - methods Prosthesis-Related Infections - diagnosis Prosthesis-Related Infections - microbiology RNA, Ribosomal, 16S - genetics Sensitivity and Specificity
Title	Diagnostic accuracy of 16S rDNA PCR, multiplex PCR and metagenomic next-generation sequencing in periprosthetic joint infections: a systematic review and meta-analysis
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