Professional Documents
Culture Documents
• Chairperson: Dr.Yashodha.H.T
• Presenter: Dr.Mohan.T.Shenoy
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HematologicIntroduction
Profile of Sepsis in Neonates:
Neutrophil CD64 as a Diagnostic Marker
Pediatrics 2008;121;129-134
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SEPSIS IN NEONATES
• Global problem and is a significant contributor to morbidity and
death.
• Most studies focus on infants of very low birth weight, given their
immature immune system and the added contribution of a variety
of risk factors.
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NEED FOR THE STUDY
It is difficult to identify neonates with sepsis before receipt of the blood
culture results.
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Acute phase reactants (for example, procalcitonin and C-reactive protein)
have also been studied as markers of neonatal sepsis.
Among the various cytokines, most studies have confirmed the utility of
Interleukin-6 as an early marker of neonatal sepsis. The cutoff values for
interleukin-6 to diagnose sepsis have ranged from 18 to 31 pg/mL.
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It is extremely important to make an early diagnosis of sepsis, because
prompt institution of antimicrobial therapy improves outcomes.
Isolation of bacteria from a central body fluid (usually blood )is the standard
and most-specific method to diagnose neonatal sepsis.
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Neutrophil surface CD64
Recently, attention has been directed to the leukocyte cell surface antigens as
diagnostic markers of sepsis.
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Technological
advances in flow
cytometry
Rapidly quantitate, with
precision, and, importantly for
neonates, with minimal blood
volumes,
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STUDY DETAILS
Authors hypothesized that neutrophil surface CD64 expression
would be a sensitive index for the detection of neonatal sepsis.
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OBJECTIVE
The goal was to determine the utility of neutrophil CD64 as a diagnostic
marker for sepsis in neonates.
METHODS
The length of antibiotic therapy varied on the basis of the severity of the
illness and the discretion of the attending neonatologist.
Complete blood count with differential, blood culture, and CD64 index
measurement were performed, and neutrophil CD64 indices were correlated
with the diagnoses of confirmed and suspected sepsis
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2 or more of the following clinical features were used to identify patients for
sepsis evaluations:
d) Hypotension; 12
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(3) Metabolic changes,
As part of the evaluation, blood was drawn for a complete blood count,
including manual differential,blood culture, and CD64 index.
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The following previously validated hematologic criteria were
used as indicators for sepsis:
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SEPSIS STRATIFICATION
A. Confirmed sepsis
B. Suspected sepsis
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CD64 Index
The first commercially available assay for PMN CD64, developed by Trillium
Diagnostics, LLC is a fluorescence based, no wash flow cytometric assay,
namely the Leuko64.
I. to CD64 and
II. to CD163,
D. Software program for automated analysis of the flow cytometric data that
reports PMN CD64 as a CD64 index.
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Briefly, 50 microL of whole blood or phosphate-buffered saline–
diluted whole blood was incubated for 10 minutes at room
temperature with a saturating amount of fluorescein
isothiocyanate-conjugated anti-CD64 monoclonal antibody or
isotype control (Leuko64 kit; TrilliumDiagnostics, Scarborough,
ME), followed by ammonium chloride-based red cell lysis.
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RESULTS
There were 10 confirmed sepsis episodes with positive culture
results:
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RESULTS
There were 293 episodes of sepsis evaluations for 163 infants.
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RESULTS
Sepsis episodes had higher neutrophil CD64 indices (5.61 vs
2.63)
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In the present study, CD64 index was found to have
the highest AUC, compared with commonly used
hematologic parameters (including band count and
immature/ total neutrophil ratio), for diagnosing
confirmed neonatal sepsis
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SUMMARY OF INFLAMMATORY
MARKERS in HEMATOLOGICAL
PROFILE
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CONCLUSION
In the present study, we found the CD64 index to have the highest
AUC, compared with commonly used hematologic parameters for
diagnosing confirmed neonatal sepsis.
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In fact, the combination of the CD64 index and the ANC criterion, without
any other hematologic indices, demonstrated 95% sensitivity for all sepsis
episodes and a 93% negative predictive value ( for predicting which
episodes did not fit septic criteria )
These findings strongly suggest that the objective CD64 index could
replace the band count (and its derived immature/total neutrophil ratio) as a
hematologic indicator of sepsis
Future studies are warranted for direct comparison of the CD64 index with
the band count and immature/total neutrophil ratio, to determine whether the
former may supplant these subjective criteria in conjunction with the ANC.
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ROLE OF CD 64 TESTING
The quantitation of CD64 is rapid (60 minutes) with the use of
flow cytometry and uses minimal blood volume.
In fact, for the present study, no extra blood was obtained from
the neonates to perform this test; the samples sent for the
complete blood count proved adequate.
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MERITS
CD64 index was elevated significantly during neonatal sepsis episodes
and was the most-diagnostic measure of confirmed sepsis;
In conjunction with the ANC, the CD64 index showed promise for
replacing the subjectively measured band count and immature/total
neutrophil ratio.
DEMERITS
Only available in specialized research labs
CD64 expression has not been studied extensively in ill preterm neonate
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SUGGESTIONS
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Statistical Analyses
Comparisons between the sepsis and no-sepsis groups were performed by using
Student’s t test or 2 analysis, as appropriate. All analyses were performed by using
GraphPad Prism 3.02 (GraphPad Software, San Diego,CA).
Receiver operating characteristic curves were generated and analyzed for the area
under the curve (AUC); pairwise comparisons were performed by using MedCalc
for Windows 9.2.0.2 (MedCalc Software, Mariakerke, Belgium).
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