Polymerase Chain Reaction Can Detect Bacterial DNA in Aseptically Loose Total Hip Arthroplasties

Clarke, M.; Roberts, C. S.; Lee, Paul T. H.; Gray, John; Keene, G; Rushton, Neil

doi:10.1097/01.blo.0000136839.90734.b7

Cited by 69 publications

(44 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Broad-range PCR is a very sensitive method and, thus, prone to false-positive results from contaminating DNA from different sources [20]. Attempts have been made to reduce the false-positive rates by introducing a detection threshold [21] but this may diminish the sensitivity of the test.…”

Section: Discussionmentioning

confidence: 99%

Diagnosis of prosthetic joint infections using UMD-Universal Kit and the automated multiplex-PCR Unyvero i60 ITI® cartridge system: a pilot study

et al. 2015

View full text Add to dashboard Cite

cultures of periprosthetic tissue samples were compared with the results of broad-range 16S rRNA gene real-time PCR (UMD-Universal Pathogen DNA Extraction and PCR Analysis, Molzym GmbH, Germany) and the multiplex-PCR Unyvero ITI ® cartridge system (U-ITI; Curetis AG, Germany). Conventional culture and broad-range 16S rRNA gene real-time PCR were performed on all samples. U-ITI was used in a subgroup of 28 cases including all culture-positive cases. The agreement of the results from the methods was assessed. Results Of 54 cases, seven were culture-positive. Broadrange 16S rRNA gene real-time PCR gave 6, U-ITI 3 concordant positive results. Of the 47 culture-negative samples, 46 were also negative by broad-range 16S rRNA gene realtime PCR resulting in a 96 % (52/54) agreement between 16S rRNA gene PCR and culture. Of the 21 culture-negative samples analysed with U-ITI, 20 gave negative results, including the single 16S rRNA gene PCR-positive/culturenegative specimen. The rate of agreement between U-ITI and culture results was 82 % (23/28). Conclusion This pilot study gave no indication of superiority of the used NAATs over conventional culture methods for the microbiological diagnosis of PJI. Drawbacks are susceptibility to contamination in the case of 16S rRNA gene real-time PCR, labour-intensive DNA extraction and limited pathogen panel in the case of the multiplex cartridge PCR system. More prospective trials are needed to evaluate the diagnostic performance of NAATs and their impact on the clinical management of PJI. Keywords

show abstract

Section: Discussionmentioning

confidence: 99%

Diagnosis of prosthetic joint infections using UMD-Universal Kit and the automated multiplex-PCR Unyvero i60 ITI® cartridge system: a pilot study

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Mariani et al, the first investigator who applied PCR for detection of bacteria in patients with joint implants, reported that 59.1% (preoperative) or 51.4% (intraoperative) of culture-negative cases were PCR positive. Clarke et al, 1 who reported that 46% (revision tissue) or 21.4% (primary tissue) of culture-negative specimens were PCR positive, noted the problem of poor specificity of a universal PCR assay compared to its high sensitivity. They also mentioned the difficulty that we too experienced, namely choosing between a less sensitive protocol with better specificity or underestimation of very low levels of bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…These limitations may be especially important for the detection of orthopedic infections, because occult infections involving prosthetic joint implants may not be detectable by conventional culture. [1][2][3] Furthermore, skeletal infections caused by Mycobacterium tuberculosis (Tb) take a long time to cultivate from clinical specimens because of the prolonged generation time of this organism. 4 Infections associated with orthopedic surgery are caused by a variety of bacteria.…”

Section: Introductionmentioning

confidence: 99%

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections

Kobayashi

Bauer

Tuohy

et al. 2006

Journal Orthopaedic Research

View full text Add to dashboard Cite

ABSTRACT:We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of ''molecular Gram stain'' with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results. ß

show abstract

“…This inflammation is characterised by macrophage infiltration and proinflammatory cytokine secretion, and the pattern of cytokines consists of tumour necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), and interleukin-6 (IL-6) [10][11][12]. After phagocytosis or the contact of the implant wear debris with the adhering bacterial components [6][7][8][9][13][14][15][16][17], the nuclear transcription factor-kappa B (NFkappa B) of the macrophage is activated, and the macrophages express the above mentioned proinflammatory cytokines in response [11,12,18]. These proinflammatory cytokines disrupt the local balance of osteogenesis and osteolysis through the proliferation of osteoclasts and the inhibition of osteoblasts [12,[19][20][21].…”

Section: Introductionmentioning

confidence: 99%

Presence of interleukin-17C in the tissue around aseptic loosened implants

Hou

Zhang

et al. 2013

International Orthopaedics (SICOT)

View full text Add to dashboard Cite

Purpose The most common long-term complication of joint arthroplasty is aseptic loosening. The proinflammatory cytokines secreted by macrophages are involved in aseptic loosening. Recently, a novel proinflammatory cytokine IL-17C was reported to participate in inflammatory diseases by synergising with proinflammatory cytokines. However, the relationship between IL-17C and the aseptic loosening is unclear. Methods The tissues around aseptic loosened implants were collected during revision surgery and handled by formalin fixation and embedded in paraffin. The presence of IL-17C in the tissues around the aseptic loosened implants was investigated in 12 aseptic loosening patients using immunofluorescence. Results The presence of IL-17C protein in the tissues around aseptic loosened implants was detected by immunofluorescence. There are no statistical differences between optical density of IL-17C in aseptic loosening samples and in rheumatoid arthritis samples (positive control). Conclusions These results suggest the presence of IL-17C in aseptic loosening. Interleukin-17C was related to the inflammation of aseptic loosening, possibly by contributing to the inflammation and osteolysis in the tissues surrounding aseptic loosened implants.

show abstract

Polymerase Chain Reaction Can Detect Bacterial DNA in Aseptically Loose Total Hip Arthroplasties

Cited by 69 publications

References 18 publications

Diagnosis of prosthetic joint infections using UMD-Universal Kit and the automated multiplex-PCR Unyvero i60 ITI® cartridge system: a pilot study

Diagnosis of prosthetic joint infections using UMD-Universal Kit and the automated multiplex-PCR Unyvero i60 ITI® cartridge system: a pilot study

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections

Presence of interleukin-17C in the tissue around aseptic loosened implants

Contact Info

Product

Resources

About