Innovative Molecular Approaches for the Diagnosis of Prosthetic Joint Infections
Trial Snapshot
- Phase
- Not Applicable
- Status
- Enrolling By Invitation
- Sponsor
- Enrollment
- 70
- Locations
- 1
- Primary Endpoint
- Concordance between analytical methods
Study Overview
Brief Summary
The goals of this observational study are:
- to evaluate the concordance between traditional method (culture) and genomic method in diagnosing prosthetic joint infections;
- to define an algorithm for laboratory diagnosis of prosthetic joint infections, which could be easily applied in our hospital setting;
- to test different DNA extraction protocols to obtain high yields of microbial nucleic acids from infected human tissue samples.
Detailed Description
Subjects A total of 70 patients from the Department of Reconstructive Surgery and Osteoarticular Infections (CRIO) and the Hip Units of IRCCS Ospedale Galeazzi -Sant'Ambrogio for suspected infection of their prosthesis will be enrolled in the study. Each sample will be analized with both traditional culture and Next Generation Sequencing (NGS).
On the basis of culture results patients will be divided into 3 groups.
Sample collection for Molecular analysis Sampling from peri-prosthetic tissue with macroscopic signs of infection will be taken from the synovial membrane, bone-prosthetic interface and intramedullary canals. Synovial fluid will be sterilely collected in DNAse and RNAse free containers for NGS analysis before performing the arthrotomy. Sterile swabs and pads will be used to collect samples from interface between tissue and bone implant and from intramedullary canal for NGS analysis. Bone tissue will be collected with new sterile instrumentation. Samples will be frozen within 2 hours from collection at -80°C until .
Sample collection for culture Sample collection of periprosthetic tissues and implant will be performed during the surgery after the proper exposure of the affected site. Synovial fluid will be collected in sterile screw cap tubes. Periprosthetic tissues and implant will be collected in sterile containers by using new sterile instrumentation. All samples will be sent to the Laboratory within 2 hours from collection. Samples after treatment with 0.1% (w:v) Dithiothreitol to dislodge bacteria from biofilm will be cultured on agar plates and broth for aerobes and anaerobes for 15 days. Microbial identification will be performed by biochemical assays.
A further tissue sample, collected in a separate sterile screw cap tube, will be used for host DNA removal to isolate microbial DNA, for shotgun NGS. Samples will be treated with saponin and/or different commercial kits for microbial DNA extraction. The dsDNA will be quantified with a Qubit fluorometer, and its purity will be determined with a nanodrop spectophotometer.
Study Design
- Study Type
- Observational
- Observational Model
- Cohort
- Time Perspective
- Prospective
Eligibility Criteria
- Ages
- 18 Years to — (Adult, Older Adult)
- Sex
- All
- Accepts Healthy Volunteers
- No
Inclusion Criteria
- •Age >18 years
- •Males and females
- •Patients with a clinical indication for knee and hip revision surgery admitted at IRCCS Ospedale Galeazzi - Sant'Ambrogio
Exclusion Criteria
- •Insufficient amount of periprosthetic tissue and synovial fluid collected
- •Patients unable to sign the Informed Consent
- •Pregnancy and breast feeding with declaration of patient
- •Age <18 years
Outcomes
Primary Outcomes
Concordance between analytical methods
Time Frame: 24 months
Concordance between methods will be analyzed by 1. Patient categorization (Infected/not infected) 2. Microorganisms identification
Secondary Outcomes
- Definition of an algorithm for laboratory diagnosis of prosthetic joint infections(36 months)