ADVIA 120 CSF Assay

ADVIA 120 CSF Assay

ADVIA 120... gets it right the 1st time,
now 1st to market with an FDA cleared
automated CSF assay
Making a positive difference to human health
.daily.

ADVIA 120 CSF Assay

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CSF Reference Values - Cell Counts

Parameter

Adults

Neonates

RBC (cells/L)

WBC (cells/L)

0-5

0 - 30

Lymphocytes (%)

60 20%

20 20%

Monocytes (%)

30 15%

70 20%

Neutrophils (%)

2 4%

4 4%

Cell Distributions in CSF samples

Normal CSF:
~ if cells present would be 100% mononuclear cells
Abnormal samples:
Neuts (PMNs)
Untreated acute bacterial infections, meningitis

Lymphs/Monos (MNs)
Viral infections, multiple sclerosis, fungal infections,
CNS* syphilis, tuberculosis

Eos
Parasitic CNS* infestations

All cell populations

Mixed infections
* Central Nervous System

CSF Sample
Current Practice
WBC and RBC
Cell Counts

WBC
Differential

Chamber Count
2-10 min

Cytospin Sample
10-15 min

High Cell Count?

Prepare Sample Dilution

Prepare Slide & Stain

Repeat

Slide Review 5-15 min

Chamber Count

Total Labor : 30 min - 45 min

ADVIA 120 CSF Assay Flow Chart

Combine
Equal Volume
CSF + Reagent

Patient CSF
Sample

CSF Reagent

Prepared Sample

Patient CSF Result:

RBC
WBC
WBC Diff

Manual
Inspect
Sample

Chamber
Counts
High
WBC?

Cytospin
Slide

Stain
Slide

Dilute
Sample

Chamber
Counts

ADVIA
120

RBC
WBC
Diff

Review
Slide

RBC
WBC
Diff

30 - 45
min

ADVIA 120
Inspect
Sample

Prepare
Sample

5 - 10
min

Current Practice
CSF
Sample
WBC
Differential

WBC and RBC

Cell Counts

r
o
b
a
L
l
Tota
Chamber Count
2-10 min

n
i
m
: 30

n
i
m
- 45

Cytospin Sample
10-15 min

High Cell Count

Dilute Sample

Repeat Chamber
Count

Prepare Slide & Stain

Review Slide
5-15 min

CSF Assay Kit Components

Start up installation kit contains:
hardware key
installs on the parallel port of the ADVIA 120 computer
accesses the proprietary CSF software

Reagent kit contains:

CSF reagent - 8 ml bottle
sufficient for 25 tests/vial

Control kit:
1 vial each of 2 levels
sufficient for 10 tests/vial

CSF Assay Kit Components

Software Installation kit contains:
Access key

Reagent kit contains:

CSF reagent
25 30 day open stability Tests

Control kit:
2 Levels Low & High
1 vial each
10 tests/vial
10 day open stability

Kit Components

Running a Sample: Routine Operations, Manual ID Screen

Running a Sample

On Board Advertisement in V3 Software

CSF Screen

CSF Sample Run Screen

monos
lymphs

neuts

eos
Research Use Only

RBCs
noise

noise

ADVIA 120 CSF Assay

Normal Sample
ADVIA 120
Mono

Neut
Eos

Lymph

Eos

RBC

Manual

# WBC

# RBC

# Neuts

# Lymphs

# Monos

# PMN

# MN

Reportable Results are...

Displayed on the Run Screen, printed & transmitted to
the LIS.
Automated WBC on all samples
Automated RBC on samples < 1500 RBCs cells/L
PMN, MN, and Differential on samples with >20 / L
(absolute and %)
WBC Differential includes:

Neutrophils
Lymphocytes
Monocytes
Eosinophils (currently for research use only, not reportable)

Reportable Results
Automated WBC on all samples
Automated RBC on samples < 1500 RBCs
cells/L
Differential, PMN and MN (absolute and %) on
samples with >20/L WBC
WBC Differential includes:
Neutrophils
Lymphocytes
Monocytes
Eosinophils (currently for research use)

ADVIA 120 CSF Assay: Abnormal

Mono

Neut

Lymph

RBC

Eos

ADVIA 120

Manual

# WBC

94

95

# RBC

298

325

% Neuts

47

52

%Lymphs

34

44

%Monos

20

%PMN

47

48

% MN

53

52

ADVIA 120 CSF Assay

Sample with High RBC
ADVIA 120
Mono

Neut

Lymph

Eos

RBC

Manual

# WBC

# RBC

284

248

# Neuts

# Lymphs

# Monos

# PMN

# MN

ADVIA 120 CSF Assay

Sample with High WBC
ADVIA 120
Mono

Neut

Lymph

Eos

RBC

Manual

# WBC

124

135

# RBC

# Neuts

31

29

# Lymphs

38

45

# Monos

47

52

# Eos

# PMN

32

38

# MN

85

97

ADVIA 120 CSF Assay

Abnormal Sample
ADVIA 120
Mono

Neut

Lymph

RBC

Eos

Manual

# WBC

94

95

# RBC

298

325

% Neuts

47

52

%Lymphs

34

44

%Monos

20

%PMN

47

48

% MN

53

52

Results

WBC Linearity
Range
0-50 / L
50 - 5,000 / L

Max. Deviation
5 cells
10%

RBC Linearity
Range
0-50 / L
50 - 1,500 / L

Max. Deviation
5 cells
10%

Within Run Precision and Accuracy (Nominal 100 cells / L; 150 samples,
at least 50% abnormal)
Parameter
Stan. Dev. / Bias
%CV / %Bias
WBC
15
15%
RBC
15
15%
# PMN
20
20%
#MN
20
20%

Summary of features
Reduces:

labor intensive manual testing

turn around time
workstations
need for specialist to perform the test

Standardizes CSF analysis

Provides Quality control for method previously uncontrolled

Extends the useful life of the prepared sample for up to 4 hours

Provides a platform for future developments
investigate feasibility of adapting method to other body fluid assays (I.e. synovial, pleural, amniotic)

Summary of Features
Reduces:
Labor intensive manual testing
Turn around time
Standardizes CSF analysis
Provides Quality Control for method previously
uncontrolled
No specialist needed to perform the test
Provides a platform for future developments
Investigating feasibility of other fluid assays
Synovial
Pleural
Amniotic

CSF Assay Performance

Characteristics

Performance Characteristics

WBC Linearity
Range
0 - 50 / L
50 - 5,000 / L

RBC Linearity
Range
0 - 50 / L
50 - 1,500 / L

Max. Deviation
5 cells
10%

Max. Deviation
5 cells
10%

Within Run Precision and Accuracy (Nominal 100 cells / L; 150 samples,
at least 50% abnormal)
Parameter
WBC
RBC
# PMN
#MN

Stan. Dev. / Bias

15
15
20
20

%CV / %Bias
15%
15%
20%
20%

ADVIA 120 CSF Assay

RBC Count
3000
r = 0.988
y = 0.93x + 5
Syx = 62
Manual Mean = 139
ADVIA 120 Mean = 135
Bias = -4

ADVIA 120 RBC

2500

2000

1500

1000

500

0
0

500

1000

1500
Manual RBC

2000

2500

3000

ADVIA 120 CSF Assay

WBC Count
3000
r = 0.988
y = 0.87x + 8
Syx = 29
Manual Mean = 55
ADVIA 120 Mean = 56
Bias = 1

ADVIA 120 WBC

2500

2000

1500

1000

500

0
0

500

1000

1500
Manual WBC

2000

2500

3000

ADVIA 120 CSF Assay

Mononuclear and Polymorphonuclear Cells

500

800
r = 0.953
y = 1.02x + 3
Syx = 16
Manual Mean = 18
ADVIA 120 Mean = 21
Bias = 3

ADVIA 120 # MN

400
350
300

r = 0.981
y = 0.81x + 2
Syx = 14
Manual Mean = 18
ADVIA 120 Mean = 17
Bias = -1

700
600
ADVIA 120 # PMN

450

250
200
150

500
400
300
200

100
100

50

0
0

100

200

300

Manual # MN

400

500

200

400

600

Manual # PMN

Comparison of manual mononucelar (MN) and polymorphonuclear (PMN) cell counts to ADVIA 120

800

ADVIA 120 CSF Assay

WBC Differential
400
r = 0.967
y = 1.23x + 0
Syx = 6
Manual Mean = 3
ADVIA 120 Mean = 4
Bias = 1

200

350
300
ADVIA 120 # Lymph

ADVIA 120 # Neut

250

200
r = 0.964
y = 0.86x + 1
Syx = 9
Manual Mean = 10
ADVIA 120 Mean = 10
Bias = 0

150

100

250

180

r = 0.928
y = 1.24x + 0
Syx = 8
Manual Mean = 4
ADVIA 120 Mean = 5
Bias = 1

160
140
ADVIA 120 # Mono

300

200
150

120
100
80
60

100
40

50

50

20

0
0

100

200

Manual # Neut

300

0
0

100

200

300

400

Manual # Lymph

Comparison of Manual Neuts, Lymph and Mono counts to ADVIA 120

50

100
Manual # Mono

150

200

6000

3000

5000

2500

4000

2000

Observed RBC

Observed WBC

ADVIA 120 CSF Assay

RBC and WBC Linearity

3000
2000
1000

1500

1000

500

1000

2000

3000

Expected WBC

4000

5000

6000

500

1000

1500
Expected RBC

2000

2500

3000

CSF Analysis Artifacts

CSF Analysis Artifacts

Signature of Oil or Fatty Substance

Manual:
ADVIA120:

RBC= 28 Cells/ L
RBC= 46 Cells/ L

WBC= 1 Cells/ L
WBC= 3 Cells/ L

CSF Analysis Artifacts

RBC Coincidence in samples with RBC counts > 1500 cells/L

Manual Result
Incorrect ADVIA 120 Result

WBC/L
25
1772

RBC/L
64300
38313

CSF Analysis Artifacts

High RBC Cell Count sample - diluted with PBS
Complete Sphering after 4 minutes incubation

Undiluted Sample

Diluted Sample with PBS

CSF Analysis Artifacts

High RBC Cell Count sample - diluted with Saline
Incomplete sphering > 4 minutes requires longer incubation

4 minute incubation
incomplete sphering

10 minute incubation
sphering complete

CSF Analysis Artifacts

Sample with RBC populations that fail to sphere completely

RBCs in Lymph Region

WBC/L

RBC/L

%Lymphs

Manual Result

17

1060

25

ADVIA 120 Result

61

650

86

CSF Analysis Artifacts

Sample containing Microcytic and/or Hypochromic RBCs

RBCs in Noise Region

WBC/L

RBC/L

Manual Result

75

1807

ADVIA 120 Result

81

1277

CSF Analysis Artifacts

Sample with Budding Yeast Forms (Circled)

WBC/L

RBC/L

%Lymphs

Manual Result

16

100

ADVIA 120 Result

40

38

93

Back up slides

Cerebrospinal Fluid (CSF)

CSF microscope slides

Normal Sample

Multiple Sclerosis

100 % mononuclear

Lymphs/Monos/PMNs

CSF Cytograms
Cell Analysis Regions
Monos
Neuts
Lymphs
RBC
Noise

Eos
Research Use Only

Noise