Identification of Molecular Marker of Coagulase-negative Staphylococci for the Diagnosis of Prosthetic Joint Infections

• ClinicalTrials.gov Identifier: NCT04661345
• Recruitment Status: Recruiting
• First Posted: December 10, 2020
• Last Update Posted: December 10, 2020
• Sponsor: Istituto Ortopedico Galeazzi
• Information provided by (Responsible Party): Istituto Ortopedico Galeazzi

Study Description

Brief Summary:

One of the major causes of prosthetic joint failure is infection. Recently, coagulase-negative Staphylococci (CoNS) have been identified as emergent, nosocomial pathogens involved in subclinical prosthetic joint infections (PJIs). The diagnosis of PJIs mediated by CoNS is complex and demanding due to the absence of clear clinical signs derived from the host immune system response. In this scenario, the key to successful surgical treatment is the capability to differentiate between aseptic implant loosening and septic failure. Hence, the central hypothesis of this study is that proteomic analysis of the secretome of CoNS clinical isolates associated with the characterization of patient synovial fluids will reveal a panel of putative biomarkers tightly linked to PJIs. The confirmation of the presence of bacterial PJI biomarkers in synovial fluids of infected patients will pave the way for the development of a new reliable test capable of aiding in the diagnosis of subclinical PJIs.

Condition or disease	Intervention/treatment
Prosthetic Joint Infection	Other: Proteomic analysis of synovial fluids; Other: Proteomic analysis of the isolated coagulase-negative Staphylococci

Detailed Description:

Task 1.1: Synovial fluids will be collected from 60 patients referred to IRCCS Galeazzi Orthopedic Institute for the revision of a septic prosthesis and from 40 patients undergoing revision due to aseptic failure. The collected synovial fluids will be stored at -80°C until analysis.

According to MSIS guidelines, the measurement of CRP and ESR levels, synovial WBC count, and the microbiological culture of the explanted device and periprosthetic tissue will be conducted following routine protocols. Only synovial fluid collected from patients of the infected group diagnosed with a PJI mediated by CoNS will be eligible for subsequent proteomic analyses. Task 1.2: The secretome of at least 10 isolates of the retrieved species of CoNS will be characterized. In particular, CoNS clinical isolates will be cultured in a drip flow reactor to induce the production of biofilm and the secretome analyzed to perform the first selection of proteins of interest.

Task 2.1: Proteins extracted from the synovial fluids of patients with or without PJI will be carbamidomethylated and digested by trypsin. Thereafter, peptide mixtures will be analyzed by nano liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS). Label-free quantifications (LFQ) will be applied to assess the levels of proteins expressed by bacteria and secreted in patient synovial fluids.

Task 2.2: Using proteomic software (i.e. MaxQuant, Perseus, etc.), the bioinformatic analysis will be performed to determine a pool of proteins of interest present only in samples belonging to the infected group of patients which would be closely related to the presence of bacteria.

Task 3.1: Proteins of interest identified in Task 1.2 and Task 2.2 as putative biomarkers related to CoNS-mediated PJIs will be selected for targeted proteomic analyses by means of additional bioinformatic analyses.

Task 3.2: To confirm the presence of putative biomarkers defined in Task 3.1 in synovial fluid of enrolled patients, targeted proteomic analysis will be performed on all the infected/not infected samples collected in Task 1.1. Prototypic peptides for each putative biomarker will be selected by Skyline software. Ionization and fragmentation behaviors of all candidate peptides will be studied.

Task 3.3: Finally, the information from the targeted proteomic analysis will be correlated to the outcomes of the diagnosis formulated following the MSIS guidelines in order to evaluate the diagnostic value of each selected biomarker.

Study Design

• Study Type: Observational
• Estimated Enrollment: 100 participants
• Observational Model: Case-Control
• Time Perspective: Prospective
• Official Title: Molecular Hallmarks of Coagulase-negative Staphylococci: Toward the Identification of Biomarkers for the Rapid Diagnosis of Subclinical Periprosthetic Joint Infections
• Actual Study Start Date: October 14, 2020
• Estimated Primary Completion Date: September 14, 2023
• Estimated Study Completion Date: October 14, 2023

Groups and Cohorts

Group/Cohort	Intervention/treatment
Septic group; The septic group is composed of patients with prosthetic joint infection mediated by coagulase-negative Staphylococci (CoNS)	Other: Proteomic analysis of synovial fluids; Characterization of the proteome of infected/aseptic synovial fluid for the identification of molecular hallmarks strictly related to the presence of a bacterial infection mediated by coagulase-negative Staphylococci; Other: Proteomic analysis of the isolated coagulase-negative Staphylococci; Characterization of the secretome of coagulase-negative Staphylococci isolated from infected prosthetic implants
Aseptic group; The aseptic group is composed of patients with implant failure unrelated to infection.	Other: Proteomic analysis of synovial fluids; Characterization of the proteome of infected/aseptic synovial fluid for the identification of molecular hallmarks strictly related to the presence of a bacterial infection mediated by coagulase-negative Staphylococci

Outcome Measures

Primary Outcome Measures :

1. Identification of molecular hallmarks strictly related to the presence of coagulase-negative Staphylococci in synovial fluids [ Time Frame: 3 years ]
   Characterization of the synovial fluids of septic/aseptic enrolled patients for the identification of a pool of protein of interest strictly related to the presence of coaglase-negative Staphylococci

Secondary Outcome Measures :

1. Proteomic analysis of coagulase-negative Staphylococci [ Time Frame: 3 years ]
   Characterization of the secretome of CoNS clinical isolate, and the characterization of the proteome of the synovial fluids of septic/aseptic enrolled patients

Biospecimen Retention:   Samples With DNA

bacterial strain

Eligibility Criteria

• Ages Eligible for Study: 18 Years and older (Adult, Older Adult)
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: No
• Sampling Method: Non-Probability Sample

Study Population

Patients subjected to a revision of the articular prosthesis due to bacterial infection or to an aseptic loosening of the implant

Criteria

Inclusion Criteria:

• ≥ 18 years old patients
• Patients subjected to a revision of the articular prosthesis due to bacterial infection mediated by coagulase-negative Staphylococci (CoNS) or to an aseptic failure of the implant
• Collection of a sufficient volume of synovial fluid
• Patient consent by signing an informed consent form

Exclusion Criteria:

• Patients not fulfilling the inclusion criteria
• Collection of a hemolytic sample

Contacts and Locations

Contacts

Contact: Elena Cittera, MSc; 00390266214057; elena.cittera@grupposandonato.it

Locations

Italy

IRCCS Istituto Ortopedico Galeazzi; Recruiting

Milan, Italy, 20161

Contact: Elena Cittera, MSc 00390266214057 elena.cittera@grupposandonato.it

Sub-Investigator: Marta Bottagisio, Ph.D.

Sponsors and Collaborators

Istituto Ortopedico Galeazzi

Investigators

• Principal Investigator: Marta Bottagisio, Ph.D.; IRCCS Istituto Ortopedico Galeazzi

More Information

• Responsible Party: Istituto Ortopedico Galeazzi
• ClinicalTrials.gov Identifier: NCT04661345
• Other Study ID Numbers: SINOVIOMA
• First Posted: December 10, 2020
• Last Update Posted: December 10, 2020
• Last Verified: December 2020

Individual Participant Data (IPD) Sharing Statement:

• Plan to Share IPD: Undecided

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

Keywords provided by Istituto Ortopedico Galeazzi:

Orthopedics; Proteomics; Synovial Fluids Biomarkers; Coagulase-negative Staphylococci

Additional relevant MeSH terms:

Infections; Communicable Diseases; Arthritis, Infectious; Disease Attributes; Pathologic Processes; Arthritis; Joint Diseases; Musculoskeletal Diseases; Coagulase; Coagulants