The detection of microbial DNA but not cultured bacteria is associated with increased mortality in patients with suspected sepsis—a prospective multi-centre European observational study

O'Dwyer, Michael J; Starczewska, ME; Schrenzel, Jacques; Zacharowski, Kai; Ecker, David J.; Sampath, Rangarajan; Brealey, David; Singer, Mervyn; Libert, Nicolas; Wilks, Martin F.; Vincent, Jean‐Louis

doi:10.1016/j.cmi.2016.11.010

Cited by 23 publications

(15 citation statements)

References 26 publications

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“…While the presence of bacterial DNA in these blood samples did not indicate the presence of viable organisms, it suggested that the clinical blood cultures were under-representing the presence of bloodstream infections in this cohort. These results are comparable to similar studies using molecular diagnostic platforms have also reported under representation of bloodstream infections when blood culture diagnostics were compared to PCR based methods [ 13 , 51 – 55 ]. This study also outlined several cases where the bacterial DNA amplified from the blood sample had good concordance with bacterial pathogens that were recovered from pertinent clinical diagnostic cultures.…”

Section: Discussionsupporting

confidence: 89%

“…However, the prevalence of polymicrobial DNA in whole blood from septic patients suggested there could be greater propensity for polymicrobial infections in sepsis than currently appreciated using cultivation-dependent and broth-enrichment based approaches. Similar results from direct blood analysis have shown utility of molecular profiling for identification of microbial DNA and its utility as an additional tool for sepsis diagnostics [ 13 , 54 , 55 ].…”

Section: Discussionmentioning

confidence: 77%

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Bacterial DNA patterns identified using paired-end Illumina sequencing of 16S rRNA genes from whole blood samples of septic patients in the emergency room and intensive care unit

et al. 2018

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BackgroundSepsis refers to clinical presentations ranging from mild body dysfunction to multiple organ failure. These clinical symptoms result from a systemic inflammatory response to pathogenic or potentially pathogenic microorganisms present systemically in the bloodstream. Current clinical diagnostics rely on culture enrichment techniques to identify bloodstream infections. However, a positive result is obtained in a minority of cases thereby limiting our knowledge of sepsis microbiology. Previously, a method of saponin treatment of human whole blood combined with a comprehensive bacterial DNA extraction protocol was developed. The results indicated that viable bacteria could be recovered down to 10 CFU/ml using this method. Paired-end Illumina sequencing of the 16S rRNA gene also indicated that the bacterial DNA extraction method enabled recovery of bacterial DNA from spiked blood. This manuscript outlines the application of this method to whole blood samples collected from patients with the clinical presentation of sepsis.ResultsBlood samples from clinically septic patients were obtained with informed consent. Application of the paired-end Illumina 16S rRNA sequencing to saponin treated blood from intensive care unit (ICU) and emergency department (ED) patients indicated that bacterial DNA was present in whole blood. There were three clusters of bacterial DNA profiles which were distinguished based on the distribution of Streptococcus, Staphylococcus, and Gram-negative DNA. The profiles were examined alongside the patient’s clinical data and indicated molecular profiling patterns from blood samples had good concordance with the primary source of infection.ConclusionsOverall this study identified common bacterial DNA profiles in the blood of septic patients which were often associated with the patients’ primary source of infection. These results indicated molecular bacterial DNA profiling could be further developed as a tool for clinical diagnostics for bloodstream infections.Electronic supplementary materialThe online version of this article (10.1186/s12866-018-1211-y) contains supplementary material, which is available to authorized users.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 77%

Bacterial DNA patterns identified using paired-end Illumina sequencing of 16S rRNA genes from whole blood samples of septic patients in the emergency room and intensive care unit

et al. 2018

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“…Thus, PCR allows for early diagnosis of sepsis and rapid treatment (antibacterial or antifungal), which lowers the treatment/hospital cost (LIESENFELD et al, 2014). Although, microbial DNA can be found even in healthy individuals, a previous study showed that the presence of microbial DNA in the blood of human patients with sepsis indicated a high risk of death (O'DWYER et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Polymerase Chain Reaction and blood culture for diagnosis of canine sepsis

et al. 2018

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Sepsis is characterized by the presence of organ dysfunction secondary to the dysregulated systemic inflammatory response associated with an infection, and has high mortality rates. Traditional diagnostic techniques based on non-microbiological isolation are time-consuming and may delay treatment. Thus, this study aimed to compare bacterial and fungal broad-range polymerase chain reaction (PCR) and blood culture for diagnosis of sepsis in dogs. Blood samples from 88 dogs with suspected sepsis were analyzed by blood culture, and PCR to detect bacterial and fungal DNA. On blood culture, 20 (22.7%) samples tested positive for bacterial isolates; however, none tested positive for fungi. Through PCR analysis, bacterial DNA was detected in 46 (52.3%) animals, whereas fungal DNA was present in one (1.1%) sample. Our results showed that PCR-based testing has important diagnostic value for canine blood infections because it has a shorter turnaround time and higher sensitivity than traditional blood culture.

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“…In another study, the same group ( 16 ) identified 84 BC-negative PCR/ESI-MS-positive microorganisms, of which 42 microorganisms (50%) had support from clinical findings. In a European ICU sepsis study ( 27 ) (a subgroup study of that described in reference 10 ), the 28-day mortality was found to be higher in patients with BC-negative PCR/ESI-MS-positive results than in patients with BC-negative PCR/ESI-MS-negative results (42% versus 26%; P = 0.001). This association with disease severity may perhaps be due to true bloodstream infection in a substantial proportion of cases with BC-negative PCR/ESI-MS-positive results.…”

Section: Discussionmentioning

confidence: 97%

Performance of PCR/Electrospray Ionization-Mass Spectrometry on Whole Blood for Detection of Bloodstream Microorganisms in Patients with Suspected Sepsis

et al. 2020

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Blood culture (BC) often fails to detect bloodstream microorganisms in sepsis. However, molecular diagnostics hold great potential. The molecular method PCR/electrospray ionization-mass spectrometry (PCR/ESI-MS) can detect DNA from hundreds of different microorganisms in whole blood. The aim of the present study was to evaluate the performance of this method in a multicenter study including 16 teaching hospitals in the USA (n=13) and Europe (n=3). First, on 2,754 contrived whole blood samples, with or without spiked microorganisms, PCR/ESI-MS produced 99.1% true positive and 97.2% true negative results. Secondly, among 1,460 patients with suspected sepsis (sepsis-2 definition), BC and PCR/ESI-MS on whole blood were positive in 14.6% and 25.6% of cases, respectively, with the following result combinations: BC+/PCR/ESI-MS-, 4.3%; BC+/PCR/ESI-MS+, 10.3%; BC-/PCR/ESI-MS+, 15.3%; and BC-/PCR/ESI-MS-, 70.1%. Compared with BC, PCR/ESI-MS showed the following sensitivities (coagulase-negative staphylococci not included): Gram-positive bacteria, 58%; Gram-negative bacteria, 78%; and Candida species, 83%. The specificities were > 94% for all individual species. Patients treated with prior antimicrobial medications (n=603) had significantly increased PCR/ESI-MS positivity rates compared with patients without prior antimicrobial treatment, 31% vs 22% (p<0.0001), with pronounced differences for Gram-negative bacteria and Candida species. In conclusion, PCR/ESI-MS showed excellent performance on contrived samples. On clinical samples, it showed high specificities, moderately high sensitivities for Gram-negative bacteria and Candida species, and elevated positivity rates during antimicrobial treatment. These promising results encourage further development of molecular diagnostics on whole blood for detection of bloodstream microorganisms in sepsis.

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The detection of microbial DNA but not cultured bacteria is associated with increased mortality in patients with suspected sepsis—a prospective multi-centre European observational study

Abstract: Presence of microbial DNA in patients with suspected sepsis might define a patient group at higher risk of death.

Cited by 23 publications

References 26 publications

Bacterial DNA patterns identified using paired-end Illumina sequencing of 16S rRNA genes from whole blood samples of septic patients in the emergency room and intensive care unit

Bacterial DNA patterns identified using paired-end Illumina sequencing of 16S rRNA genes from whole blood samples of septic patients in the emergency room and intensive care unit

Polymerase Chain Reaction and blood culture for diagnosis of canine sepsis

Performance of PCR/Electrospray Ionization-Mass Spectrometry on Whole Blood for Detection of Bloodstream Microorganisms in Patients with Suspected Sepsis

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