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

2019 ◽  
Vol 4 (2) ◽  
pp. 224-228
Author(s):  
Michael A Lane ◽  
Neeraja Ganeshraj ◽  
Alice Gu ◽  
David K Warren ◽  
Carey-Ann D Burnham
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Diagnostic Yield ◽  
Diagnostic Tools ◽  
Rrna Gene ◽  
Joint Infections ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections ◽  
Culture Positive ◽  
Culture Negative

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.

scholarly journals Bacterial diversity in suspected prosthetic joint infections: an exploratory study using 16S rRNA gene analysis

2012 ◽  
Vol 65 (2) ◽  
pp. 291-304 ◽  
Author(s):  
Yijuan Xu ◽  
Vibeke Børsholt Rudkjøbing ◽  
Ole Simonsen ◽  
Christian Pedersen ◽  
Jan Lorenzen ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Diversity ◽  
Exploratory Study ◽  
Gene Analysis ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Joint Infections ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections

scholarly journals Detection of Prosthetic Hip Infection at Revision Arthroplasty by Immunofluorescence Microscopy and PCR Amplification of the Bacterial 16S rRNA Gene

1999 ◽  
Vol 37 (10) ◽  
pp. 3281-3290 ◽  
Author(s):  
Michael M. Tunney ◽  
Sheila Patrick ◽  
Martin D. Curran ◽  
Gordon Ramage ◽  
Donna Hanna ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Immunofluorescence Microscopy ◽  
Joint Infection ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Bacterial Dna ◽  
Prosthetic Joint ◽  
Culture Positive ◽  
Bacterial 16S Rrna Gene

In this study the detection rates of bacterial infection of hip prostheses by culture and nonculture methods were compared for 120 patients with total hip revision surgery. By use of strict anaerobic bacteriological practice during the processing of samples and without enrichment, the incidence of infection by culture of material dislodged from retrieved prostheses after ultrasonication (sonicate) was 22%. Bacteria were observed by immunofluorescence microscopy in 63% of sonicate samples with a monoclonal antibody specific forPropionibacterium acnes and polyclonal antiserum specific for Staphylococcus spp. The bacteria were present either as single cells or in aggregates of up to 300 bacterial cells. These aggregates were not observed without sonication to dislodge the biofilm. Bacteria were observed in all of the culture-positive samples, and in some cases in which only one type of bacterium was identified by culture, both coccoid and coryneform bacteria were observed by immunofluorescence microscopy. Bacteria from skin-flake contamination were readily distinguishable from infecting bacteria by immunofluorescence microscopy. Examination of skin scrapings did not reveal large aggregates of bacteria but did reveal skin cells. These were not observed in the sonicates. Bacterial DNA was detected in 72% of sonicate samples by PCR amplification of a region of the bacterial 16S rRNA gene with universal primers. All of the culture-positive samples were also positive for bacterial DNA. Evidence of high-level infiltration either of neutrophils or of lymphocytes or macrophages into associated tissue was observed in 73% of patients. Our results indicate that the incidence of prosthetic joint infection is grossly underestimated by current culture detection methods. It is therefore imperative that current clinical practice with regard to the detection and subsequent treatment of prosthetic joint infection be reassessed in the light of these results.

scholarly journals Role of Universal 16S rRNA Gene PCR and Sequencing in Diagnosis of Prosthetic Joint Infection

2011 ◽  
Vol 50 (3) ◽  
pp. 583-589 ◽  
Author(s):  
M. Marin ◽  
J. M. Garcia-Lechuz ◽  
P. Alonso ◽  
M. Villanueva ◽  
L. Alcala ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Prosthetic Joint Infection ◽  
Joint Infection ◽  
Rrna Gene ◽  
Prosthetic Joint

scholarly journals Prosthetic joint infection by Bordetella holmesii: Case report and a review of the literature

2020 ◽  
Vol 43 (11) ◽  
pp. 748-750
Author(s):  
Mariana Fernandez-Pittol ◽  
Jordi Bosch ◽  
Laura Morata ◽  
Luis Lozano ◽  
Juan Carlos Martínez Pastor ◽  
...  
Keyword(s):  
Case Report ◽  
Synovial Fluid ◽  
Antibiotic Treatment ◽  
Knee Prosthesis ◽  
Case Reports ◽  
Joint Effusion ◽  
Rrna Gene ◽  
Joint Infections ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections

Introduction: Bordetella holmesii is a Gram-negative coccobacillus involved in different infections mostly described in case reports. Prosthetic joint infections in relation to this pathogen are rare. Here, we present the third case of B. holmesii in a patient without anatomical or functional spleen dysfunction. Case report: The patient was a 62-year-old female with a total knee prosthesis implanted in 1997 that required multiple replacements of the femoral component due to aseptic loosening in the past years. The patient was admitted to our hospital for an elective replacement surgery due to new radiological signs of loosening. B. holmesii was isolated from synovial fluid obtained during surgery. The identification was performed by matrix-assisted laser desorption ionization–time of flight mass spectrometry and confirmed by 16S rRNA gene amplification and sequencing. Antibiotic treatment was started but 14 days after surgery the patient presented pain and joint effusion. An arthrocentesis was performed and synovial fluid culture was positive again for B. holmesii. Surgical debridement including polyethylene replacement was performed and antibiotic treatment was continued for 3 months. After a 2-year follow-up period, the patient remained asymptomatic and physical examination showed normal function of the prosthesis. Conclusion: B. holmesii is an uncommon cause of bone and joint infections. This case indicates that this microorganism is a potential pathogen of prosthetic or native arthritis, and it should be considered when cultures are negative and in cases presenting torpid evolution.

scholarly journals 69016s rRNA Gene PCR: An Unreliable Tool for the Diagnosis of Prosthetic Joint Infections (PJI)

2014 ◽  
Vol 1 (suppl_1) ◽  
pp. S195-S195
Author(s):  
Michael A. Lane ◽  
Carey-Ann Burnham ◽  
Alice P. Gu ◽  
Neeraja Ganeshraj ◽  
Qian Liu ◽  
...  
Keyword(s):  
Rrna Gene ◽  
Joint Infections ◽  
Prosthetic Joint ◽  
Prosthetic Joint Infections

scholarly journals First Experience of a Multicenter External Quality Assessment of Molecular 16S rRNA Gene Detection in Bone and Joint Infections

2014 ◽  
Vol 53 (2) ◽  
pp. 419-424 ◽  
Author(s):  
Chloé Plouzeau ◽  
Pascale Bémer ◽  
Anne Sophie Valentin ◽  
Geneviève Héry-Arnaud ◽  
Didier Tandé ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Joint Infection ◽  
Rrna Gene ◽  
Gene Detection ◽  
External Quality ◽  
Multicenter Studies ◽  
Joint Infections ◽  
Significant Difference ◽  
Bead Mill

The objective of this study was to assess the performance of seven French laboratories for 16S rRNA gene detection by real-time PCR in the diagnosis of bone and joint infection (BJI) to validate a large multicenter study. External quality control (QC) was required owing to the differences in extraction procedures and the molecular equipment used in the different laboratories. Three proficiency sets were organized, including four bacterial DNA extracts and four bead mill-pretreated osteoarticular specimens. Extraction volumes, 16S rRNA gene primers, and sequencing interpretation rules were standardized. In order to assess each laboratory's ability to achieve the best results, scores were assigned, and each QC series was classified as optimal, acceptable, or to be improved. A total of 168 QCs were sent, and 160 responses were analyzed. The expected results were obtained for 93.8%, with the same proportion for extracts (75/80) and clinical specimens (75/80). For the specimens, there was no significant difference between manual and automated extraction. This QC demonstrated the ability to achieve good and homogeneous results using the same 16S rRNA gene PCR with different equipment and validates the possibility of high-quality multicenter studies using molecular diagnosis for BJI.

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