Detection of bacterial DNA in serial synovial samples obtained during antibiotic treatment from patients with septic arthritis

Heijden, I. M. Van Der; Wilbrink, Bert; Vije, Arianne E. M.; Schouls, Leo M.; Breedveld, Ferdinand C.; Tak, Paul P.

doi:10.1002/1529-0131(199910)42:10<2198::aid-anr23>3.0.co;2-n

Cited by 86 publications

(42 citation statements)

References 15 publications

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“…Both live and dead bacteria may result in positive PCR assays (17,25). Despite negative bacterial cultures, the presence of bacterial DNA in clinical specimens has been shown for prolonged periods in septic arthritis (6,44), pulmonary tuberculosis (14,49), and leptospirosis (3,27). The significance of the persistence of DNA for long periods after antibiotic treatment is uncertain (17,25).…”

Section: Discussionmentioning

confidence: 99%

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PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

et al. 2005

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We used broad-range PCR amplification and sequencing to detect and identify bacterial DNA in 156 valves of patients treated for infective endocarditis (IE). Bacterial DNA was found more frequently in patients who underwent valve replacement while on antibiotic treatment for IE (60%) than in patients who had completed antibiotic treatment for IE (37%; P ‫؍‬ 0.02). We found specific bacterial DNA in valves removed from 11 of 30 patients who had completed antibiotic treatment for IE. Six had no histological evidence of IE. The presence of DNA was significantly correlated with the presence of histologic lesions (P ‫؍‬ 0.001) and with the presence of bacteria detected by Gram staining (P < 0.001). Bartonella and streptococci were detected for much longer after antibiotic treatment by PCR than other species (P ‫؍‬ 0.047 and 0.04, respectively), and coagulase-negative staphylococci were detected for much shorter periods (P ‫؍‬ 0.02). The finding that bacterial DNA was more likely to be detected in valves of patients with active IE than in patients who had completed antibiotic treatment for IE shows that bacterial DNA is cleared slowly. There was no significant correlation between the duration of antibiotic therapy and the presence of bacterial DNA in valves. Since the persistence of bacterial DNA in valves does not necessarily indicate the persistence of viable bacteria, the detection of bacterial DNA in valves from IE patients should be interpreted with caution, in particular in those patients with a past history of treated IE.

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Section: Discussionmentioning

confidence: 99%

“…In patients with septic arthritis, bacterial DNA can be found in synovial tissue for long periods after cultures become negative (44). The persistence of bacterial DNA despite effective antibiotic treatment has also been demonstrated in patients with pulmonary tuberculosis (14,49) and leptospirosis (3,27).…”

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PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

et al. 2005

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“…However, PCR (polymerase chain reaction), based on molecular techniques, has led to an increase in diagnosis of bacterial etiologies for clinical specimens with negative bacterial culture (12,13) ids of patients (14)(15)(16)(17). In this study, we aimed at detecting Staphylococcal enterotoxin E in synovial fluids of patients with rheumatoid arthritis.…”

Section: Introductionmentioning

confidence: 99%

Molecular Assay of Staphylococcal Enterotoxin E in Synovial Fluid of Patients with Rheumatoid Arthritis

et al. 2018

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Background: The role of Staphylococcal enterotoxin E (Superantigen E) in rheumatoid arthritis pathogenesis has been considered. This paper aimed at determining Staphylococcal enterotoxin E in the synovial fluid of rheumatoid arthritis patients. Methods: In this study, 100 synovial fluid samples of patients with rheumatoid arthritis were examined. The primers pairs were designed based on the S. baureus enterotoxin type E (entE) gene, GenBank: M21319.1. All samples were subjected to DNA extraction separately. Then, polymerase chain reaction (PCR) was implemented. Data were analyzed by descriptive statistics. Results:The PCR results indicated that Staphylococcal enterotoxin E gene existed in synovial fluid samples of 25% of patients with rheumatoid arthritis, with a high percentage. Conclusions:The study results revealed that a high percentage of patients with rheumatoid arthritis have Staphylococcal enterotoxin type E gene in their synovial fluid. However, further studies are needed to assess other Staphylococcal enterotoxins. This finding may provide a model for diagnosing rheumatoid arthritis disease. The results of this study have presented some evidence regarding endogenous origin of involved superantigen in patients with rheumatoid arthritis.

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“…The use of PCR amplification of 16S rRNA gene has been proposed for broad-range detection of eubacteria in synovial fluid (17,18). However, nearly all broad-based PCR assays reported thus far involve laborious time-consuming postamplification processing (e.g., gel electrophoresis, Southern blotting, or sequencing), making them impractical for routine clinical use (8,14,17,18).…”

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confidence: 99%

“…However, nearly all broad-based PCR assays reported thus far involve laborious time-consuming postamplification processing (e.g., gel electrophoresis, Southern blotting, or sequencing), making them impractical for routine clinical use (8,14,17,18). For example, to date, the largest recent study using broad-based real-time PCR study for diagnosis of SA demonstrated high sensitivity and specificity but relied on sequencing for definitive pathogen identification (5).…”

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Rapid PCR-Based Diagnosis of Septic Arthritis by Early Gram-Type Classification and Pathogen Identification

et al. 2008

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Septic arthritis (SA) is a rheumatologic emergency associated with significant morbidity and mortality. Delayed or inadequate treatment of SA can lead to irreversible joint destruction and disability. Current methods of diagnosing SA rely on synovial fluid analysis and culture which are known to be imprecise and time-consuming. We report a novel adaptation of a probe-based real-time PCR assay targeting the 16S rRNA gene for early and accurate diagnosis of bacterial SA. The assay algorithm consists of initial broad-range eubacterial detection, followed by Gram typing and species characterization of the pathogen. The platform demonstrated a high analytical sensitivity with a limit of detection of 10 1 CFU/ml with a panel of SA-related organisms. Gram typing and pathogen-specific probes correctly identified their respective targets in a mock test panel of 36 common clinically relevant pathogens. One hundred twenty-one clinical synovial fluid samples from patients presenting with suspected acute SA were tested. The sensitivity and specificity of the assay were 95% and 97%, respectively, versus synovial fluid culture results. Gram-typing probes correctly identified 100% of eubacterial positive samples as to gram-positive or gram-negative status, and pathogen-specific probes correctly identified the etiologic agent in 16/20 eubacterial positive samples. The total assay time from sample collection to result is 3 h. We have demonstrated that a real-time broad-based PCR assay has high analytical and clinical performance with an improved time to detection versus culture for SA. This assay may be a useful diagnostic adjunct for clinicians, particularly those practicing in the acute care setting where rapid pathogen detection and identification would assist in disposition and treatment decisions.Septic arthritis (SA) is a rheumatologic emergency associated with significant morbidity and mortality (6, 9). Delayed or inadequate treatment of SA can lead to irreversible joint destruction with subsequent disability. Accordingly, prompt diagnosis and early initiation of therapy are critical in improving the outcome (7).The diagnosis of SA in the acute care setting is challenging because of the relatively poor sensitivity and specificity of clinical examination findings and lack of a rapid reliable diagnostic assay. Further, overreliance on conventional laboratory tests for synovial fluid analysis is hindered by the relatively poor performance characteristics of these methods (11,12,16). In particular, the sensitivity of Gram staining has been reported in the range of 29% to 50% (3, 4), and the sensitivity of culture may be only 82% (9). Lack of a rapid and accurate diagnostic tool results in acute care clinicians often choosing the conservative approach of hospital admission and empirical broadspectrum antibiotics for patients with suspected SA. The benefits of this management strategy may be offset, however, by added costs and potential iatrogenic complications associated with unnecessary treatment and hospitalizations, as well ...

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Detection of bacterial DNA in serial synovial samples obtained during antibiotic treatment from patients with septic arthritis

Abstract: PCR analysis can be used to monitor the presence of bacterial DNA in synovial samples from patients with septic arthritis during antibiotic treatment. The absence of bacterial DNA could help in the decision to discontinue antibiotic treatment.

Cited by 86 publications

References 15 publications

PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

Molecular Assay of Staphylococcal Enterotoxin E in Synovial Fluid of Patients with Rheumatoid Arthritis

Rapid PCR-Based Diagnosis of Septic Arthritis by Early Gram-Type Classification and Pathogen Identification

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