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Diagnostic utility of broad range bacterial 16S rRNA gene PCR with degradation of human and free bacterial DNA in bloodstream infection is more sensitive than an in-house developed PCR without degradation of human and free bacterial DNA.

Rogina P, Skvarc M, Stubljar D, Kofol R, Kaasch A - Mediators Inflamm. (2014)

Bottom Line: Broad range 16S rRNA gene PCR was performed from whole blood (SepsiTest) or blood plasma (IHP) and compared to blood culture results.SepsiTest identified more relevant pathogens than blood cultures (P = 0.008); in three patients (13%) results from blood culture and SepsiTest were congruent, whereas in four cases (17.4%) relevant pathogens were detected by SepsiTest only.A diagnostic algorithm using CD64 index as a decision maker to perform SepsiTest shows improved detection of pathogens in patients with suspected blood stream infection and may enable earlier targeted antibiotic therapy.

View Article: PubMed Central - PubMed

Affiliation: Novo Mesto General Hospital, 8000 Novo Mesto, Slovenia.

ABSTRACT
We compared a commercial broad range 16S rRNA gene PCR assay (SepsiTest) to an in-house developed assay (IHP). We assessed whether CD64 index, a biomarker of bacterial infection, can be used to exclude patients with a low probability of systemic bacterial infection. From January to March 2010, 23 patients with suspected sepsis were enrolled. CD64 index, procalcitonin, and C-reactive protein were measured on admission. Broad range 16S rRNA gene PCR was performed from whole blood (SepsiTest) or blood plasma (IHP) and compared to blood culture results. Blood samples spiked with Staphylococcus aureus were used to assess sensitivity of the molecular assays in vitro. CD64 index was lower in patients where possible sepsis was excluded than in patients with microbiologically confirmed sepsis (P = 0.004). SepsiTest identified more relevant pathogens than blood cultures (P = 0.008); in three patients (13%) results from blood culture and SepsiTest were congruent, whereas in four cases (17.4%) relevant pathogens were detected by SepsiTest only. In vitro spiking experiments suggested equal sensitivity of SepsiTest and IHP. A diagnostic algorithm using CD64 index as a decision maker to perform SepsiTest shows improved detection of pathogens in patients with suspected blood stream infection and may enable earlier targeted antibiotic therapy.

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Related in: MedlinePlus

Flow diagram of patients enrolled in the study and results from CD64 index, BC, and ST (SIRS = systemic inflammatory response syndrome, ST = SepsiTest, and BC = blood culture).
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fig1: Flow diagram of patients enrolled in the study and results from CD64 index, BC, and ST (SIRS = systemic inflammatory response syndrome, ST = SepsiTest, and BC = blood culture).

Mentions: In the clinical study, 61 patients were screened for SIRS criteria in the emergency department. 23 of the patients met the inclusion criteria and were enrolled in the study (Figure 1). All patients were subsequently transferred to an intensive care unit for supportive treatment. Enrolled patients were between 24 and 88 years old (median age 59 years). Systemic bacterial infection was suspected in 19 patients (82.6%); four patients (17.4%) with other reasons for SIRS were identified: three patients suffered from a myocardial infarct and one patient from thyrotoxicosis. All but one of the patients with suspected systemic bacterial infection showed an elevated CD64 index (Figure 1).


Diagnostic utility of broad range bacterial 16S rRNA gene PCR with degradation of human and free bacterial DNA in bloodstream infection is more sensitive than an in-house developed PCR without degradation of human and free bacterial DNA.

Rogina P, Skvarc M, Stubljar D, Kofol R, Kaasch A - Mediators Inflamm. (2014)

Flow diagram of patients enrolled in the study and results from CD64 index, BC, and ST (SIRS = systemic inflammatory response syndrome, ST = SepsiTest, and BC = blood culture).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4120914&req=5

fig1: Flow diagram of patients enrolled in the study and results from CD64 index, BC, and ST (SIRS = systemic inflammatory response syndrome, ST = SepsiTest, and BC = blood culture).
Mentions: In the clinical study, 61 patients were screened for SIRS criteria in the emergency department. 23 of the patients met the inclusion criteria and were enrolled in the study (Figure 1). All patients were subsequently transferred to an intensive care unit for supportive treatment. Enrolled patients were between 24 and 88 years old (median age 59 years). Systemic bacterial infection was suspected in 19 patients (82.6%); four patients (17.4%) with other reasons for SIRS were identified: three patients suffered from a myocardial infarct and one patient from thyrotoxicosis. All but one of the patients with suspected systemic bacterial infection showed an elevated CD64 index (Figure 1).

Bottom Line: Broad range 16S rRNA gene PCR was performed from whole blood (SepsiTest) or blood plasma (IHP) and compared to blood culture results.SepsiTest identified more relevant pathogens than blood cultures (P = 0.008); in three patients (13%) results from blood culture and SepsiTest were congruent, whereas in four cases (17.4%) relevant pathogens were detected by SepsiTest only.A diagnostic algorithm using CD64 index as a decision maker to perform SepsiTest shows improved detection of pathogens in patients with suspected blood stream infection and may enable earlier targeted antibiotic therapy.

View Article: PubMed Central - PubMed

Affiliation: Novo Mesto General Hospital, 8000 Novo Mesto, Slovenia.

ABSTRACT
We compared a commercial broad range 16S rRNA gene PCR assay (SepsiTest) to an in-house developed assay (IHP). We assessed whether CD64 index, a biomarker of bacterial infection, can be used to exclude patients with a low probability of systemic bacterial infection. From January to March 2010, 23 patients with suspected sepsis were enrolled. CD64 index, procalcitonin, and C-reactive protein were measured on admission. Broad range 16S rRNA gene PCR was performed from whole blood (SepsiTest) or blood plasma (IHP) and compared to blood culture results. Blood samples spiked with Staphylococcus aureus were used to assess sensitivity of the molecular assays in vitro. CD64 index was lower in patients where possible sepsis was excluded than in patients with microbiologically confirmed sepsis (P = 0.004). SepsiTest identified more relevant pathogens than blood cultures (P = 0.008); in three patients (13%) results from blood culture and SepsiTest were congruent, whereas in four cases (17.4%) relevant pathogens were detected by SepsiTest only. In vitro spiking experiments suggested equal sensitivity of SepsiTest and IHP. A diagnostic algorithm using CD64 index as a decision maker to perform SepsiTest shows improved detection of pathogens in patients with suspected blood stream infection and may enable earlier targeted antibiotic therapy.

Show MeSH
Related in: MedlinePlus