Comparison of cultures and 16S rRNA sequencing for identification of bacteria in two-stage revision arthroplasties: preliminary report

Abstract

BackgroundThe use of a prefabricated spacer in two-stage revision arthroplasty remains one of the few surgery strategies for infected-joint arthroplasty treatment, despite the many unidentified microorganisms in the infected joint replacements reported in some recent studies. The aim of this prospective survey was to investigate if the sonication followed by polymerase chain reaction (PCR) can improve bacterial identification on the surfaces of prefabricated spacers and if the systemic laboratory mediators of infection and positive microbiological results can take a role of predictive factors of infection and clinical failures in 2-years follow-up.MethodsThirteen patients with prosthetic joint infection were investigated. Bacterial culture and deoxyribonucleic acid (DNA) sequencing were used to detect bacteria on the surface of prefabricated spacers removed during the second stage of revision arthroplasty. The results of pre- and intraoperative culture and DNA sequencing were compared. Minimum follow-up was 2 years.ResultsThe result of tissue cultures in second-stage revision arthroplasties revealed positive results in 15 % of patients with Coagulase-negative Staphylococci (CNS) growth. Bacterial DNA was found in over 90 % of patients with negative synovial fluid culture. Positive PCR results revealed potential pathogenic bacteria and species of human and environmental microflora with low virulence. Clinical failures at final follow-up were recorded in 2 (16.6 %) patients.ConclusionThe lack of clinical signs of infection, negative culture of preoperative joint aspirate, and intraoperative specimens do not exclude the presence of bacteria on the surfaces of spacers. The positive results of sonication and molecular tests should be interpreted as real pathogenicity factors in the light of the clinical and laboratory data, especially for patients with immunodeficiency. We confirmed our previous results that sonication followed by PCR and sequencing improved bacterial identification.

Diagnosis Of Persistent Infection In Prosthetic Two-Stage Exchange: PCR analysis of Sonication fluid From Bone Cement Spacers

“…Indeed, PCR cannot confirm the presence of viable bacteria in the samples 10 . As PCR is a very sensitive method, it can also show contamination 11 .…”

Propidium monoazide–polymerase chain reaction for detection of residual periprosthetic joint infection in two-stage revision

“…In the last decade, PCR has been evaluated in diagnosis of PJI in an attempt to overcome the limited sensitivity of culture methods [7][8][9]. While PCR is a rapid and relatively sensitive technique, it can falsely indicate the presence of residual infection at stage two revisions after antibiotic treatment due to dead bacteria.…”

Sonication fluid culture of antibiotic-loaded bone cement spacer has high accuracy to confirm eradication of infection before reimplantation of new prostheses

“…Nine studies [ 17 , 19 , 24 – 30 ] also provided the culture results of intraoperative tissue of bone cement spacers. Five [ 17 , 19 , 21 , 28 , 29 ] studies used a prospective design and the others [ 20 , 24 – 27 , 30 ] a retrospective cohort design. The QUASDAS-2 scores of qualities ranged from 9 to 11 with a mean of 10.27.…”