Lack of Additional Diagnostic Yield of 16s rRNA Gene PCR for Prosthetic Joint Infections

Abstract

Introduction Medical management of prosthetic joint infections (PJIs) relies on the identification of causative organisms through traditional culture-based approaches to guide therapy. However, diagnosis of many PJIs remains challenging, with many clinically apparent infections remaining culture-negative. Molecular diagnostics have the potential to increase diagnostic yield, particularly among culture-negative PJIs. Methods Bone, tissue, or synovial fluid from patients with clinically identified PJIs were collected for inclusion in this study. Samples were assessed with traditional cultures and classified as culture-positive or -negative after 48 h. Samples subsequently underwent a Staphylococcus aureus-/Kingella kingae-specific PCR followed by a 16s rRNA gene PCR. Results A total of 77 unique patients with clinically identified PJIs contributed a total of 89 samples for inclusion in the study. There were 54 culture-negative and 35 culture-positive samples evaluated. The sensitivity and specificity of S. aureus PCR in culture-positive samples was 57.1% (95% CI, 34.1%–78.1%) and 92.9% (95% CI, 66.1%–98.9%), respectively. Among culture-positive samples, 16s rRNA gene PCR correctly identified 3 of 21 (14.3%) samples with S. aureus and 2 of 5 (40%) samples with Streptococcus spp. All molecular tests were negative in those with clinically identified, culture-negative PJI. Conclusions Our study suggests that these diagnostic tools have a limited role in PJI diagnosis.

The current trend for prosthetic joint infection diagnosis from culture to molecular: a literature review

“…when using the 16S rRNA as the primer. 25 Hence, the clinical use of multiplex PCR and broad primer PCR currently limited to identify rare pathogens, when the inflammation was low, or when the culture was REVIEW negative. 20,21,26- 28 The latest trend in molecular methods is next-generation sequencing.…”

Clinical Use of a 16S Ribosomal RNA Gene-Based Sanger and/or Next Generation Sequencing Assay to Test Preoperative Synovial Fluid for Periprosthetic Joint Infection Diagnosis

“…Cultures of synovial fluid are recommended for preoperative diagnosis of PJI and pathogen identification, though they have been shown to have a relatively low sensitivity across numerous studies ( 7 , 8 ). Molecular methods such as 16S rRNA (rRNA) gene PCR followed by Sanger sequencing have been shown to have lower sensitivity than synovial fluid cultures ( 9 ), possibly due to technical limitations of Sanger sequencing, such as the inability to differentiate polymicrobial samples and a restricted limit of detection. In recent years, next-generation sequencing (NGS) has delivered metagenomic sequencing-based approaches for diagnosis of PJI; these approaches are of particular interest for culture-negative infections ( 10 – 12 ).…”

Protein detection based on rolling circle amplification sensors

“…PCR-based technologies have been vastly exploited in all areas of research, [1][2][3][4][5][6][7][8] however these technologies need advanced and expensive instrumentation. [9,10] A short DNA strand can be extended using DNA polymerases and techniques such as rolling circle amplification (RCA).…”