Diagnostic Microbiology and Infectious Disease 75 (2013) 130134

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Diagnostic Microbiology and Infectious Disease

journal homepage: www.elsevier.com/locate/diagmicrobio

Routine use of a real-time polymerase chain reaction method for detection of

bloodstream infections in neutropaenic patients,
Michela Paolucci a, Marta Stanzani b, Fraia Melchionda c, Giulia Tolomelli b, Gastone Castellani d,
Maria Paola Landini a, Stefania Varani a, Russell E. Lewis e, Vittorio Sambri a,
a
b
c
d
e

Unit of Microbiology, Department of Specialistic, Diagnostic and Experimental Medicine, University of Bologna, 40138 Bologna, Italy
Institute of Hematology Lorenzo e Ariosto Sergnoli Sant'Orsola-Malpighi Hospital, University of Bologna, 40138 Bologna, Italy
Paediatric Oncology and Hematology Unit Lalla Sergnoli, University of Bologna, 40138 Bologna, Italy
Physics Department, Bologna University, 40127 Bologna, Italy
Texas Medical Center, University of Houston College of Pharmacy, Houston, TX 77030, USA

a r t i c l e

i n f o

Article history:
Received 24 July 2012
Received in revised form 2 October 2012
Accepted 14 October 2012
Available online 22 November 2012
Keywords:
Bloodstream infection
Neutropaenia
Blood culture
PCR
Haematologic cancer patient
Antibiotic therapy

a b s t r a c t
We examined the performance of a real-time polymerase chain reaction (PCR) test (SeptiFast) for early
detection of bloodstream infection in febrile neutropaenic patients. Blood samples from 201 patients were
screened for pathogens by blood culture and by PCR on the rst day of fever. PCR results were available earlier
(median 3 days for bacteria, 5 days fungal pathogens; P 0.01). The sensitivity (0.74) and specicity (0.96) of
the PCR test were acceptable for Gram negatives when culture was considered the gold standard, but
sensitivity of the test was poorer for Gram-positive organisms (0.39). The PCR assay also led to 22.9% of invalid
results. SeptiFast speeds the microbiological diagnosis of bloodstream infection in neutropaenic patients.
However, the frequent failure of instrumental control procedures, the relatively poor sensitivity of the test,
and the lack of phenotypic data on antimicrobial susceptibility associated with its high costs suggest that this
assay cannot replace the blood cultures.
2013 Elsevier Inc. All rights reserved.

1. Introduction
Neutropaenia is a major risk factor for bloodstream infections
(BSIs) in patients with haematological cancer. Fever develops in 65%
of cancer patients receiving uoroquinolone prophylaxis during the
neutropaenic period, but a microbiologically documented diagnosis
is made in only 22% of these cases (bacteraemia 18%) (Bucaneve
et al., 2005).
Bloodstream infections are routinely diagnosed with blood
cultures taken at the onset of fever (Hughes, 2005; Penack et al.,
2006). However, one disadvantage of this method is the turnaround time of 26 days before the results are available (Bucaneve
et al., 2005). Additionally, the sensitivity of blood culture is reduced
in neutropaenic patients with haematologic malignancies because
they often receive prophylactic antibiotics and are at risk for infections caused by cell-wall decient bacteria and lamentous fungi,

Funding: This work was supported by RFO 2009-2010 from the University of
Bologna (to VS, MPL and SV); the Italian Ministry of Education, University, and Research
MIUR (PRIN 2007, MPL, VS, SV, GC).
Transparency declarations: none to declare.
Corresponding author. Tel.: +39-051-6363013.
E-mail address: vittorio.sambri@unibo.it (V. Sambri).
0732-8893/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.diagmicrobio.2012.10.012

which are rarely detected by blood culture (Carrigan et al., 2004;

Woo et al., 2001).
The detection of microbial DNA in blood by polymerase chain
reaction (PCR) is a promising approach for diagnosing BSIs
(Mancini et al., 2008). A common limitation in the assessment of
novel molecular methods is the absence of a gold standard for
detection of BSIs. In neutropaenic cancer patients in particular,
the interpretation of PCR results is limited by negative blood
culture results due to antibiotic treatment (Peters et al., 2004).
Consequently, some authors have recommended that positive PCR
results for blood culturenegative febrile episodes be interpreted
based on corresponding clinical features of the infection rather
than on purely microbiological results (Nakamura et al., 2010;
Peters et al., 2004).
Previously, we assessed the clinical utility of a commercially
available multiplex real-time PCR assay (LightCycler SeptiFast Test
M GRADE; Roche Diagnostics, Mannheim, Germany) for the microbiological diagnosis of BSIs in 100 severely immunocompromised
patients (Varani et al., 2009). Since then, we have routinely used
this procedure along with standard blood culture, evaluating in total
201 neutropaenic patients over the past 2 years. Based on this
experience and on our analysis, we have identied advantages and
limitations of this technology for diagnosis of infections in neutropaenic patients.

M. Paolucci et al. / Diagnostic Microbiology and Infectious Disease 75 (2013) 130134

2. Patients and methods

2.1. Study patients, settings, and denitions
In this prospective interventional study, we analysed 437 blood
samples from 339 consecutive febrile episodes, obtained between
June 1, 2008, and March 31, 2010. Blood samples were drawn from
201 severely neutropaenic (absolute neutrophil count b500/mm 3)
patients (23 children and 178 adults) with haematological malignancies (105 acute myeloid leukaemia, 23 acute lymphoblastic leukaemia, 34 lymphoma, 15 multiple myeloma, and 8 chronic
myeloproliferative disorders), severe aplastic anaemia (4 patients),
solid tumours (9 patients), or other disorders (2 cases of autoimmune
thrombocytopaenia, 1 case of haemophagocytic lymphohistiocytosis).
Patients were admitted to the Institute of Haematology and the
Paediatric Oncology and Haematology Unit, St. Orsola-Malpighi
University Hospital, Bologna, Italy. All patients enrolled were febrile
(38.5 C), neutropaenic, and received antibiotic and antifungal
prophylaxis, as specied by in-house guidelines. Specically, all adults
received antibiotic prophylaxis with uoroquinolones, and 91
patients who underwent allogeneic hematopoietic stem cell transplantation also received uconazole as antifungal prophylaxis. All
other patients received itraconazole or posaconazole. Paediatric
patients received cotrimoxazole and, for those who were undergoing
allogeneic haematopoietic stem cell transplantation (n = 4),
prophylaxis with uoroquinolones and uconazole.
Clinical and microbiological data were used to judge the clinical
relevance of a positive PCR or blood culture result by the attending
physician caring for the patient. Specically, coagulase-negative
staphylococci (CoNS) or Streptococcus spp. identied by culture or
PCR were not considered to be true pathogens in persistently
neutropaenic patients if a) the neutropaenic patient defervesced in
the absence of antibiotic therapy targeting the specic microorganism
and/or b) only 1 set of blood cultures tested positive in concomitance
of a PCR-negative nding.
Test samples were collected from the patients at the onset of the
febrile episode and before empirical antibiotic therapy was
administered. Blood was drawn from peripheral veins in adults. In
paediatric patients, blood was drawn from central venous catheters
(CVC), as venous access is often difcult in this population (Hall and
Lyman, 2006).
2.2. Microbiological methods
All blood samples were processed at the hospital microbiology
unit. For culture, blood was collected twice at the onset of fever within
a 30-min interval, while 3 samples were taken if CVC was present. Five
to 10 mL of blood was put into each aerobic and anaerobic culture
bottle (BacT/Alert 3D system, BioMerieux Italia, Florence, Italy),
according to the Clinical and Laboratory Standards Institute protocol
(Wilson et al., 2007). The cultures were incubated up to 132 h before
assessing a negative result. Positive blood cultures were smeared and
Gram stained. Subcultures were simultaneously started by seeding on
both nutritive and selective agar medium to obtain isolated colonies.
The isolated bacteria were biochemically identied (Vitek2 instruments and panel, BioMerieux), and antimicrobial susceptibility
testing (AST) was performed by automated instruments, as follows:
Gram-positive bacteria were tested using the Sensititre Aris system
(Trek, Cleveland, OH, USA), and Gram-negative bacteria were tested
with the Vitek2 instrument (BioMerieux).
For PCR testing, 3 mL of blood was sampled and processed for
patients who weighed 45 kg and 1.5 mL for those who were b45 kg.
Specimens were collected once at the onset of the febrile episode and
then processed by LightCycler SeptiFast Test M GRADE as described
(Varani et al., 2009). Samples for blood culture and PCR testing were
collected through a single venipuncture.

131

The PCR assay is based on 3 principal steps: 1) specimen

preparation by mechanical lysis and purication of DNA from whole
blood; 2) real-time PCR amplication of target DNA in 3 parallel
reactions (Gram-positive bacteria, Gram-negative bacteria, fungi);
and 3) detection by specic hybridization probes and automated
identication of species and controls, as described (Lehmann et al.,
2008). A dened volume of internal control was introduced into each
specimen to verify the amplication reaction. Target bacterial (Grampositive and Gram-negative) and fungal DNA were simultaneously
amplied as reagent controls. When amplication of internal control
or reagent control failed, the results of PCR were considered invalid.
Blood cultures were accepted for automated incubation from
8:00 a.m. to 7:00 p.m. (Monday to Saturday). PCR was performed
once per day from Monday to Friday (samples were received by
12:00 p.m.). Blood samples intended for PCR analyses arriving to
the laboratory after 12:00 p.m. were stored at 4 C until the next
PCR session was programmed (time limit for storage was 72 h, as
suggested by the manufacturer).
This study was conducted according to the regulations of the St.
Orsola-Malpighi University Hospital Ethical Committee.
2.3. Statistical analysis
The sensitivity, specicity, positive predictive value (PPV), and
negative predictive value (NPV) plus 95% condence intervals (CI)
were calculated for the PCR assay using blood culture results as a gold
standard reference excluding nonevaluable PCR test results. The
percentage of positive tests was compared in 2-by-2 contingency
tables using the chi-quare of Fisher's test. Time to positivity for PCR
and culture results was dened as the time interval from blood
collection to the complete identication of germs, including AST data.
The distribution of time to positivity for PCR versus cultures results
was compared using t test or MannWhitney test, when appropriate.
A 2-sided P value of b 0.05 was considered statistically signicant.
Analysis was performed using the SPSS 20 Statistical package (IBM,
Armonk, NY, USA).
3. Results
Of the 437 samples evaluated by blood culture and PCR, 100
(22.9%, corresponding to 75 febrile episodes) yielded a technically
invalid result by PCR because of the failure of the internal control or
reagent control to amplify and were excluded from further analysis.
The remaining 264 febrile episodes (corresponding to 337 samples)
were studied. The microorganisms identied are summarized in
Table 1.
In the 49 febrile episodes that were positive by both blood culture
and PCR, organisms were detected on average 2.5 days earlier by PCR
versus blood culture (P b 0.01); in 159 febrile episodes, PCR yielded a
negative result 5 days before blood culture (P b 0.01) (Fig. 1). The
overall concordance between the 2 tests was 79% (Table 2).
With blood cultures as the gold standard, the PPV of PCR assay
ranged from 0.39 (95% CI 0.0.250.53) for Gram positive, 0.74 (95% CI
0.530.88) for Gram negatives to 50% (95% CI 0.030.97) for fungi, and
67% (0.130.98) for mixed infections (Table 2). The relatively poor
sensitivity of the test for Gram-positive pathogens (0.39, 95% CI 0.25
0.53) reected the occasional failure of the PCR test to detect CoNS
growing in culture. However, CoNS may reect contamination during
venipuncture rather than true infection, especially in cases where
neutropaenic patients defervesced without antimicrobial therapy
active against CoNS. The PCR test appeared to be more useful for
detecting the presence of Gram-negative pathogens, with a more
acceptable sensitivity of 74%. Estimates of the sensitivity, specicity,
PPV, and NPV were less precise for fungi and mixed infections, given
the small number of cases (Table 2).

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M. Paolucci et al. / Diagnostic Microbiology and Infectious Disease 75 (2013) 130134

Table 1
Comparative identication of bacteria and fungi in 264 febrile episodes in neutropaenic patients.
Real-time PCR identication
Gram-positive (n = 28)

Gram-negative (n = 30)

Blood culture results

Gram positive
CoNS (12)
S. aureus (1)
(n = 49)

E. coli/CoNS (2)

Fungi (n = 2)

Mixed (n = 11)

Negative (n = 193)

ND

A. baumannii + CoNS (1) f

Klebsiella spp.+ Enterobacter
spp./CoNS (1) g

CoNS (14)
S. aureus (1)

Enterococcus spp. (4)

Streptococcus spp. (2)a

Gram negative
(n = 27)

ND

Fungi (n = 2)
Mixed (n = 3)

CoNS/Fusarium spp.b (1)

ND

Negative
(n = 181)

CoNS (7)
S. aureus (1)

E. coli (14) e
Klebsiella spp. (1)
P. aeruginosa (4)
Stenotrophomonas
maltophilia (1)
ND
E. coli + Corynebacterium
jeikeium b (1)
E. coli (5)
Klebsiella spp.(1)
Enterobacter spp.(1)

ND

E. coli + CoNS (1)

C. parapsilosis (1)
ND

ND
C. parapsilosis + CoNS (1)
E. coli + CoNS (1)
S. aureus + Klebsiella spp. (1)
Enterococcus spp. + CoNS + E. coli (1)
Streptococcus spp. + P. aeruginosa (1)
S. aureus + C. tropicalis (1)
Enterococcus spp. + CoNS (1)
CoNS + S. aureus (1)

A. fumigatus (1)

Streptococcus spp. (7)

Enterococcus spp. (1)
Corynebacterium spp. b (1)
Leuconostoc spp. b (1)
S. pluranimalium b (1)
E. coli (3)
P. aeruginosa (2)
A. lwofi b (1)

C. parapsilosis (1)
CoNS + C. glabrata (1)
ND (159)

ND = No growth or reaction detected.

a
S .mitis detected by PCR; S. pneumoniae detected by blood culture in 1 episode.
b
Microorganism not detectable by PCR test.
c
CoNS detected by SeptiFast; Fusarium sp. detected by blood culture.
d
E. coli detected by PCR; CoNS detected by blood culture in both episodes.
e
E. coli detected by PCR; K. oxytoca detected by blood culture in 1 episode.
f
SeptiFast, but not blood culture, detected mixed infection.
g
Klebsiella spp. and Enterobacter spp. detected by SeptiFast; CoNS detected by blood culture.

In 14 febrile episodes, PCR identied a Gram-negative bacterium

that did not grow in culture (Table 1). In 13 of these cases, the
identied microorganisms were judged clinically relevant. While in 1
of these 14 cases, Klebsiella pneumoniae/oxytoca was identied by PCR,
but the fever resolved without any antimicrobial therapy. This patient
was screened for other possible infection sites and no positive result
conrmed the microorganism that was detected by PCR.
With regard to fungal infections, PCR detected 1 febrile episode of
Aspergillus fumigatus and 1 episode of Candida tropicalis, for which
blood cultures were negative (Table 1). These patients were treated
immediately with targeted antifungal therapy, and the infections
resolved. We also identied 13 febrile episodes caused by polymicrobial infections; 9 were detected only by PCR and were caused by
life-threatening microorganisms (S. aureus, Enterococcus spp., Escherichia coli, Acinetobacter baumannii, Enterobacter spp., Pseudomonas
aeruginosa, Klebsiella spp., and C. tropicalis) (Dorn et al., 1976).
In 34 febrile episodes, PCR did not detect any microorganism,
while blood culture gave a positive result (Table 1). In 7 febrile
episodes, PCR failed to identify microorganisms that provoke severe
infections in neutropaenic patients, such as E. coli (n = 3), P.
aeruginosa (n = 2), Candida parapsilosis (n = 1), and Candida glabrata
(n = 1). In 21 additional febrile episodes, Gram-positive bacteria
grew in culture. The negative PCR nding in these cases was
consistent with clinical evidence indicating that CoNS and Streptococcus spp. detected by blood culture were irrelevant in 9 cases and 2
cases, respectively. In 10 cases, the clinical ndings were consistent
with an infection by Gram-positive bacteria, as the fever resolved only
after appropriate Gram-positive antibiotic coverage. Finally, in 6
samples, PCR failed to identify relevant microorganisms causing
sepsis (Acinetobacter lwofi, n = 1; Corynebacterium spp., n = 2;
Fusarium spp., n = 1; Leuconostoc spp., n = 1; Streptococcus
pluranimalium, n = 1), reecting the intrinsic limitation of the test
to detect these relatively uncommon pathogens.

In 22 febrile episodes, PCR identied microbial DNA, while the rst

peripheral blood sample was negative by blood culture. Extending the
comparison to the second or third blood sampling for cultures, we
found that, in 8 cases, the PCR results were conrmed by blood
culture, identifying CoNS (n = 6), E. coli (n = 1), and Enterobacter spp.
(n = 1) in the CVC blood sample.
In 5 febrile episodes, PCR and blood cultures generated discordant
microbial ndings (Table 1). In 1 case, S. pneumoniae was detected by
PCR and Streptococcus mitis by peripheral and catheter blood sample
cultures. As previously reported (Lehmann et al., 2008), the viridans
streptococci group can be erroneously identied by PCR as S.
pneumoniae because of a high degree of target sequence homology.
In another case, E. coli was detected by PCR and K. oxytoca was
detected by blood culture; because no other positive cultures were
available for this patient, we could not determine which test correctly
identied the infectious agent. In 3 febrile episodes, PCR detected
Gram-negative bacteria (E. coli in 2 and K. pneumoniae/oxytoca and
Enterobacter cloacae/aerogenes in 1), and the blood cultures were
positive for CoNS; the positive outcome after treatment against Gramnegative bacteria suggested that the correct results were generated
by PCR.
4. Discussion
In this study, we used a real-time, multiplex PCR assay daily in a
large cohort of neutropaenic patients with suspected BSI in a routine
setting, and the results were available in time to guide therapeutic
decisions. We also evaluated a larger number of febrile episodes for a
longer period of time than in previous studies (Lamoth et al., 2010;
von Lilienfeld-Toal et al., 2009) and identied advantages and
limitations of using this multiplex PCR assay to diagnose BSIs.
First, PCR signicantly reduced the turnaround time of the
microbiological diagnosis of BSIs in neutropaenic patients, which

M. Paolucci et al. / Diagnostic Microbiology and Infectious Disease 75 (2013) 130134

A
positive results

40

Gram positive
median 1 vs. 4 days
P < 0.001

30
20
10
0
0

20

positive results

Gram negative
median 1 vs. 4 days
P < 0.001

15
10
5
0
0
10

positive results

Fungal

median 1 vs. 6 days

P = 0.01

6
4
2
0
0

Time to result (days)

PCR
Culture
Fig. 1. Distribution of the time to positive tests for culture versus real-time PCR.
Concordant real-time PCR (black bars) and culture (grey bars) are shown for singleorganism infections. (A) Gram-positive; (B) Gram-negative; and (C) fungal organisms.
Distributions were compared using a 2-tailed MannWhitney test.

could encourage more pathogen-driven antimicrobial therapy

earlier in treatment.
PCR was also useful in identifying BSIs caused by fungiespecially
Candida species other than albicansand by lamentous fungi of the
genus Aspergillus. These ndings are relevant because early appropriate treatment is essential in reducing the mortality among highrisk patients (Morrell et al., 2005; Richardson, 2005).
PCR also detected polymicrobial bacteraemias and a single test
displayed respectable performance for the detection of Gram-

133

negative organisms, which are associated with high mortality rates

in neutropaenic patients (Klastersky and Aoun, 2004).
However, we found a major limitation of this PCR assay, i.e., the
frequent failure of instrumental control procedures, which occurred in
22.9% of samples. This problem was reported to the manufacturer, but
no technical explanation for this high failure rate could be found other
than the constitutive weakness of the method, likely due to the high
complexity of the reactions. Moreover, in 6 febrile episodes, PCR failed
to detect E. coli, P. aeruginosa, C. parapsilosis, or C. glabrata
microorganisms that can provoke severe infections in neutropaenic
patients (Raad et al., 2007). False-negative results were also obtained
in other studies (Bravo et al., 2011; Tsalik et al., 2010; von LilienfeldToal et al., 2009; Wallet et al., 2010; Westh et al., 2009). Lamoth et al.
(2010) hypothesized that part of these failures could be explained
considering the smaller blood volume used for PCR compared with
blood culture.
In 10 other cases, PCR failed to identify true infections by Grampositive bacteria; as suggested by Bloos et al. (2010), SeptiFast can fail
in detecting these organisms because Gram-positive bacterial lysis is
more difcult. Furthermore, the relatively low sensitivity of SeptiFast
for CoNS and streptococci is possibly associated with the semiquantitative analytical cut-off value that was arbitrarily established by
the manufacturer to distinguish contamination from infection in nonneutropaenic patients (Lehmann et al., 2008). Moreover, because of
the intrinsic inability of the test to detect certain microbes, PCR failed
to identify relevant microorganisms causing sepsis in 6 samples.
Overall, the PCR method led to false-negative results in 23 cases
(8%). Thus, a negative nding by PCR does not exclude the presence
of a BSI in neutropaenic patients. Neutropaenic patients exhibit
weak phagocytic activity, and even a low microbial load could be
enough to trigger a sepsis syndrome. It is possible that, in some
patients receiving antibacterial prophylaxis, the initial microbial
load at the onset of fever was below the limit of reliable detection
for the multiplex PCR assay, but sufcient to cause clinical infection
in these patients. It has been hypothesized also that the low
detection rate for microbial DNA could be related to the lack of
neutrophils containing microbial DNA in neutropaenic patients (von
Lilienfeld-Toal et al., 2009). Adjustment of the analytical aspects of
PCR may be warranted, or additional sampling may be required to
improve its sensitivity in neutropaenic patients.
One PCR-positive case (i.e., detection of K. pneumoniae/oxytoca)
was considered to be a false-positive nding; samples from other
possible infection sites did not conrm the PCR positivity in blood, and
the febrile episode resolved without any antimicrobial treatment.
These ndings underscore another limitation of the PCR method: PCRpositive/culture-negative results cannot differentiate microorganisms
in the blood that are rendered nonviable by ongoing antibiotic
treatment or from small amounts of cell-free DNA released from
remote sites of infection or microbial colonization.
The analysis of blood culture/PCR discordant results has been
performed on the basis of clinical outcome, including response to
antimicrobial therapy. Despite the limitations of this analysis, we

Table 2
Real-time PCR test performance analysed by blood culture result.
Blood culture results

Sensitivitya (95% CI)

Specicitya (95% CI)

Positive predictive valuea (95% CI)

Negative predictive valuea (95% CI)

Gram positive (n = 49)

Gram negative (n = 27)
Fungi (n = 2)
Mixed pathogens (n = 3)
All pathogens (n = 81)

0.39
0.74
0.50
0.67
0.52

0.96
0.96
0.99
0.97
0.89

0.67 (0.480.83)
0.67 (0.470.82)
0.5 (0.030.97)
0.18 (0.030.52)
0.59 (0.470.70)

0.87
0.97
0.99
0.99
0.86

(0.250.53)
(0.530.88)
(0.030.97)
(0.130.98)
(0.410.63)

(0.920.98)
(0.920.98)
(0.971.00)
(0.930.98)
(0.840.92)

(0.820.91)
(0.940.99)
(0.971.00)
(0.971.00)
(0.810.90)

Inclusion of non-technically valid PCR results in the analysis (n = 100) as negatives was also analysed by blood culture result: Gram positive: sensitivity (0.26, 0.170.38),
specicity (0.97, 0.940.98), PPV (0.68, 0.470.83), NPV (0.83, 0.790.87). Gram negative: sensitivity (0.54, 0.030.97), specicity (0.99, 0.981.0), PPV (0.5, 0.020.97), NPV (0.95,
0.920.97). Fungi: sensitivity (0.50, 0.170.38), specicity (0.97, 0.940.98), PPV (0.68, 0.470.83), NPV (0.99, 0.981.0). Mixed pathogens: sensitivity (0.67, 0.130.98), specicity
(0.97, 0.950.99), PPV (0.18, 0.030.52), NPV (0.99, 0.981.0). All pathogens: sensitivity (0.37, 0.280.46), specicity (0.91, 0.870.94), PPV (0.59, 0.470.70), NPV (0.80, 0.760.84).
a
Calculations were performed for technically valid PCR results only (n = 337).

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M. Paolucci et al. / Diagnostic Microbiology and Infectious Disease 75 (2013) 130134

could not consider any of the 2 techniques as infallible and completely

rely on it as a gold standard for diagnosis of BSIs. For example, blood
culture can be falsely negative if sampled after antimicrobial therapy
(Peters et al., 2004), while PCR can be intrinsically unable to pick up
microorganisms of dened aetiology (Lehmann et al., 2008).
We conclude that PCR signicantly speeds the microbiological
diagnosis of BSIs in neutropaenic patients and helps to identify fungal
and polymicrobial bacteraemias. However, the frequent failure of
instrumental control procedures, the false-negative results of this PCR
assay, as well as the inability to generate phenotypic data on
antimicrobial susceptibility with the current test platform, suggest
that SeptiFast cannot replace the blood cultures. Therefore PCR tests
are best used in conjunction with culture, to detect serious and
potentially multidrug-resistant pathogens earlier so clinicians can
make pre-emptive antibiotic modications until full antibiotic
susceptibility data are conrmed by standard methods.
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