Evaluation of pathogen detection from clinical samples by real-time polymerase chain reaction using a sepsis pathogen DNA detection kit

Yanagihara, Katsunori; Kitagawa, Yuko; Tomonaga, Masao; Tsukasaki, Kunihiro; Kohno, Shigeru; Seki, Masafumi; Sugimoto, Hisashi; Shimazu, Takeshi; Tasaki, Osamu; Matsushima, Asako; Ikeda, Yasuo; Okamoto, Shinichiro; Aikawa, Naoki; Hori, Shingo; Obara, Hideaki; Ishizaka, Akitoshi; Hasegawa, Naoki; Takeda, Junzo; Kamihira, Shimeru; Sugahara, Kazuyuki; Asari, Seishi; Murata, Mitsuru; Kobayashi, Yoshio; Ginba, Hiroyuki; Sumiyama, Yoshinobu; Kitajima, Masaki

doi:10.1186/cc9234

Cited by 93 publications

(90 citation statements)

References 20 publications

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“…In contrast, in the case of blood culture, antibiotic administration reduced the probability of pathogen isolation by half. This finding is consistent with results reported by other researchers 8,21,33 and supports the usefulness of SeptiFast in cases where antibiotic therapy has already been started and shows its advantages over blood cultures.…”

Section: Discussionsupporting

confidence: 83%

Microbiological Diagnosis of Sepsis: Polymerase Chain Reaction System Versus Blood Cultures

Suberviola

Márquez-López

Castellanos-Ortega

et al. 2015

American Journal of Critical Care

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Objective To compare the utility of a multiplex polymerase chain reaction system (SeptiFast) and blood cultures for detecting bacteria and fungi in blood samples from patients with severe sepsis or septic shock. Methods In a prospective observational study, whole blood samples for SeptiFast testing and for culture were collected on admission from all patients with severe sepsis or septic shock admitted to the intensive care unit between July 2011 and September 2012. SeptiFast results were compared with blood and other culture results. Results The probability of at least 1 microorganism being isolated at 6 hours was 13-fold higher with the SeptiFast test than with blood cultures (relative risk, 13.5; 95% CI, 5.05-36.06). Unlike culture results, SeptiFast test results were not associated with previous antibiotic consumption. The median time to the first positive blood culture result was 17 hours; SeptiFast results were available in 6 hours. SeptiFast detected genetic material from potentially multiresistant microorganisms in patients whose blood cultures showed no growth at all. Conclusions The SeptiFast test provided quicker microbiological diagnosis and identified significantly more microorganisms than blood cultures did, particularly when samples were collected after antibiotic therapy had started or infections were due to resistant bacteria and yeast.

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Section: Discussionsupporting

confidence: 83%

Microbiological Diagnosis of Sepsis: Polymerase Chain Reaction System Versus Blood Cultures

Suberviola

Márquez-López

Castellanos-Ortega

et al. 2015

American Journal of Critical Care

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“…It is a relatively rapid molecular method allowing detection of sepsis pathogens in approximately 6 h. There are several studies to date about diagnostic feasibility and potential clinical utility of SF [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24]. Fewer studies have evaluated this assay in routine clinical practice regarding impact on therapy [25,26].…”

Section: Discussionmentioning

confidence: 99%

Diagnostic performance and therapeutic impact of LightCycler SeptiFast assay in patients with suspected sepsis

Herne¹,

Nelovkov²,

Kutt³

et al. 2013

European Journal of Microbiology and Immunology

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Rapid and reliable identification of pathogens is very important in the management of septic patients. We retrospectively evaluated the diagnostic accuracy and clinical utility of a multiplex real-time polymerase chain reaction (PCR) assay (SeptiFast (SF)) in patients with suspected sepsis in a tertiary care hospital in Tallinn, Estonia. A total of 160 blood samples from 144 patients were included in the study. SF results were compared with corresponding blood culture (BC) results. The concordance between SF and BC was 78.8%. The rate of positive results was significantly higher in SF than in BC (33.7% vs. 21.2%, respectively; p < 0.001). A total of 27 samples were found positive by both SF and BC, 27 by SF only, and seven by BC only. Of a total of 83 microorganisms detected SF identified 71, and BC 42 (p < 0.001). SF detected markedly more patients with candidemia: 11 patients were detected by SF compared to four patients by BC. Antimicrobial treatment was changed in 21 (38.9%) of 54 SF positive cases. In conclusion, our results demonstrated the high diagnostic accuracy of SF in detection of sepsis pathogens. In conjunction with its impact on therapeutic decisions, SF proved to be a useful adjunct to conventional blood culture in the diagnosis of sepsis etiology.

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“…The targets for amplification are the internal transcribed sequences between the bacterial 16S and 23S genes, and the fungal 18S and 5.6S rRNA genes (Lehmann et al, 2008). In several studies it has been demonstrated that this test can be more sensitive than blood culture for the diagnosis of sepsis in different settings (Mancini et al, 2008;Yanagihara et al, 2010), especially in patients already receiving antibiotics (Vince et al, 2008). Moreover, in nonsepsis infections, such as bacterial endocarditis, joint or other deep infections, it is often difficult to isolate bacterial agents from clinical specimens, especially if patients undergo antibiotic treatment.…”

Section: Introductionmentioning

confidence: 99%

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

Mencacci

Leli

Cardaccia

et al. 2011

Journal of Medical Microbiology

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Early detection of aetiological agents is pivotal for adequate therapy for bacterial infections. Although culture is still considered the mainstay for laboratory diagnosis, it often lacks sensitivity, especially in patients already treated with antibiotics. The present study investigated the potential clinical utility of the commercial real-time-PCR-based system SeptiFast (SF), originally intended for diagnosis of sepsis from blood specimens, in the aetiological diagnosis of other bacterial infections, in patients undergoing antibiotic therapy. A total of 53 non-blood specimens were analysed for microbial pathogen detection by conventional culture and with SF real-time PCR: 19 (35.8 %) synovial fluids, 9 (17.0 %) cardiac valve tissues and 25 (47.2 %) purulent exudates from various body sites. Overall, the number of specimens positive for a pathogen by SF (26/53; 49.1 %) was significantly greater (P50.001) than that of specimens positive by culture (10/53; 18.9 %). In particular, SF was superior to culture for pathogen detection in cardiac valve tissues and synovial fluids. The analysis of concordance showed a fair agreement between the two methods (kappa value50.314; 95 % confidence interval50.531-0.097). Even with the limitation of the low number of specimens, this study confirmed the great potential of diagnosing bacterial infections by a molecular approach, and indicates that the real-time PCR SF system can be used for specimens other than blood, from patients undergoing antibiotic treatment.

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Evaluation of pathogen detection from clinical samples by real-time polymerase chain reaction using a sepsis pathogen DNA detection kit

Cited by 93 publications

References 20 publications

Microbiological Diagnosis of Sepsis: Polymerase Chain Reaction System Versus Blood Cultures

Microbiological Diagnosis of Sepsis: Polymerase Chain Reaction System Versus Blood Cultures

Diagnostic performance and therapeutic impact of LightCycler SeptiFast assay in patients with suspected sepsis

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

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