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ANALIS

CEofix™ SPE kit for Beckman Coulter P/ACE™ MDQ series - GB: Instructions for use

QUOTATION
INFORMATION
QUOTATION
INFORMATION
DESCRIPTION
DESCRIPTION

1. General

Date: 23-Nov-05 - Revision: 0
EURO PN A26217
ANALIS PN 10-004750
100 tests per kit

2. Intended use

The kit is intended for the electrophoretic separation of human serum proteins with the Beckman Coulter P/ACE MDQ.

3. Principle of the test

The serum proteins are separated inside a capillary under the influence of an electrical field. Differences in charge to mass ratio give different migration times.
Proteins are detected at 214 nm with a UV detector mounted on the capillary.

Clinical Significance: The electrophoretic separation of serum proteins is a universal protein-screening technique for the detection of monoclonal gammopathies and as an aid in the diagnosis of pathophysiologic states associated with changes in various protein quantities.

4. Content

Conditioner 1 x 50 mL Sodium Hydroxide 0.2 mol/L (PN. 10-004751))
Buffer 1 x 60 mL TRIS/Taurine buffer, pH 9.7 (PN 10-004752)
Rinse 1 x 45 mL Aqua bidistillata (PN 10-004753)
Initiator 1 x 14 mL Tris buffer, pH 9.7 (PN 10-004754)

5. Configuration of instrument

-BeckmanCoulter P/ACE MDQ
-UV detector and sample cooling
-P/ACE 32 Karat software version 5.0 or 7.0 (use configuration without CAESAR integration)

6. Other material needed

- Vials 2 mL (PN 144980)
- red caps (PN 144648)
- 500 µL Microvials ( PN 970668) for samples and blue caps (PN 144649)
- Blank Cartridge assembly 100 x 800 µm (PN 144738)
- Pipettes
- Capillary 25µM ID x 30 cm total length (EuroPN: 844111023 or PN 10-004788)

7. Training

The operator should be familiar with instrument (P/ACE MDQ) operation and maintenance.

8. Reagent preparation and storage

Reagents are ready to use. Store kit and buffers, in closed container, between 8°C and 30°C until expiration date.

Between operation buffer vials should be capped to avoid evaporation.

If the kit arrived damaged a replacement of the kit should be requested.

The kit contains no hazardous components.

9. Type of specimen

Biological fluids should be collected in the manner normally used for any clinical laboratory test.

Freshly drawn serum from a fasting individual is the preferred specimen.

10. Instrument preparation

figure 1


1. .

figure 2

2.

figure 3

3.

figure 4

4.

figure 5

5.

figure 6

6.

figure 7

7.

figure 8

8.

figure 9

9. Peak / Group Tables - UV - 214 nm / Use area percent

Name

ID</

Mig. Time

MT Window

Ref. ID #

EOF

1

1.7

0.06

38

EOF

2

1.75

0.06

38

EOF

3

1.8

0.06

38

EOF

4

1.85

0.06

38

EOF

5

1.9

0.06

38

gamma

6

1.95

0.06

38

gamma

7

2

0.06

38

gamma

8

2.05

0.06

38

gamma

9

2.1

0.06

38

gamma

10

2.15

0.06

38

gamma

11

2.2

0.06

38

gamma

12

2.25

0.06

38

gamma

13

2.3

0.06

38

beta

14

2.35

0.06

38

beta

15

2.4

0.06

38

beta

16

2.45

0.06

38

beta

17

2.5

0.06

38

beta

18

2.55

0.06

38

beta

19

2.6

0.06

38

alpha 2

20

2.65

0.06

38

alpha 2

21

2.7

0.06

38

alpha 2

22

2.75

0.06

38

alpha 2

23

2.8

0.06

38

alpha 2

24

2.85

0.06

38

alpha 2

25

2.9

0.06

38

alpha 2

26

2.95

0.06

38

alpha 2

27

3

0.06

38

alpha 2

28

3.05

0.06

38

alpha 2

29

3.1

0.06

38

alpha 1

30

3.15

0.06

38

alpha 1

31

3.2

0.06

38

alpha 1

32

3.25

0.06

38

alpha 1

33

3.3

0.06

38

alpha 1

34

3.35

0.06

38

albumin

35

3.4

0.06

38

albumin

36

3.45

0.06

38

albumin

37

3.5

0.06

38

albumin

38

3.55

1

0

albumin

39

3.6

0.06

38

albumin

40

3.65

0.06

38

albumin

41

3.7

0.06

38

albumin

42

3.75

0.06

38

albumin

43

3.8

0.06

38

albumin

44

3.85

0.06

38

albumin

45

3.9

0.06

38

albumin

46

3.95

0.06

38

albumin

47

4

0.06

38

albumin

48

4.05

0.06

38

albumin

49

4.1

0.06

38

albumin

50

4.15

0.06

38

albumin

51

4.2

0.06

38

albumin

52

4.25

0.06

38

albumin

53

4.3

0.06

38

albumin

54

4.35

0.06

38

albumin

55

4.4

0.06

38

albumin

56

4.45

0.06

38

albumin

57

4.5

0.06

38

Peaks can be grouped by name by entering in the  Group Def. (+)

Name

Ref. ID#

ISTD.ID#

Groupe Type

Group Def.

Unit

Analysis Channel

Quantitation

EOF

0

0

Named Peaks

(+)

UV - 214nm

Area

gamma

0

0

Named Peaks

(+)

UV - 214nm

Area

beta

0

0

Named Peaks

(+)

UV - 214nm

Area

alpha2

0

0

Named Peaks

(+)

UV - 214nm

Area

alpha1

0

0

Named Peaks

(+)

UV - 214nm

Area

albumin

0

0

Named Peaks

(+)

UV - 214nm

Area

Or download program:
spe.met ( 32 Karat Software 7.0)

Burn the capillary ends to eliminate the polyimide coating on 2mm.

11. Sample preparation

Use fresh neat serum, put 200 to 400 µL in microvial, cap with blue caps and place in outlet 48 sample holder.

12. Operations

Every day operation:

- clean instrument and interface manifold
- put new rinse, conditioner, initiator buffer vials
- put clean and dry caps on buffer vials
- run  cond_spe.met when system was down for several days (see method).
- prepare samples 200 to 400 µL of neat serum in 500 microvials
- run  spe.met  method
End of day:

- remove samples
- remove all buffer vials
Placing new capillary

-When the capillary is broken or when the capillary is not operating properly replace the capillary: according the instrument manual.
-run "cond_spe.met for conditioning

13. Calibration

No calibration is required for the kit.

Instrument should be maintained and calibrated according to Beckman Coulter specifications.

14. Analysis of results

see example

Integration windows should be corrected for a adequate grouping of fractions.

15. Calculation

No calculations are required for the qualitative, visual interpretation of CEofix SPE when comparing electropherograms to a known reference pattern or commercially available control.
The relative percent of each protein fraction is calculated, using the area of each fraction, by the instrument software.

16. Imprecision

Two normal samples and one abnormal sample was run according to "NCCLS EP5-A Preliminary Precisions Test"

N=12 Mean and %CV

Gamma Beta Alpha 2 Alpha 1 Albumine
Normal 1 13.80
3.15
7.52
1.83
13.44
1.35
5.01
2.47
59.80
1.03
Normal 2 6.96
4.39
7.77
2.64
13.10
1.22
5.54
4.79
65.37
0.77
High gamma 23.77
2.06
8.9
3.74
12.85
2.36
4.69
3.42
49.08
1.19

17. Results

The SPE clinical laboratory test is a screening procedure. Interpretation of the SPE electropherogram should always be in conjunction with a clinical evaluation because the SPE pattern is neither specific nor diagnostic by itself. Immunofixation electrophoresis is recommended to rule out or confirm the presence of monoclonal immunoglobulin on any patient.

Reference interval:
Each laboratory should establish its own reference range to conform to patient population.

For a normal population we obtained following results:


Reference interval was obtained by analyzing 133 apparently healthy male and female adults from Belgium using "non-parametric reference interval" (Analyse-it).

Protein fraction 95% Lower Limit (90% CI) 95% Upper Limit (90% CI)
Gamma 8.1 (7.3 - 8.9) 15.6 (14.6 - 16.4)
Beta 6.9 (6.8 - 7.6) 11.6 (11.2 - 13.9)
Alpha 2 9.1 (6.3 - 10.7) 16.7 (16.0 - 19.4)
Alpha 1 3.4 (2.6 - 3.6) 7.2 (6.9 - 7.4)
Albumine 53.7 (53.3 - 55.3) 64.6 (63.8 - 65.6)

18. Interference

- Plasma will produce an extra, fibrinogen peak in the gamma region.

- High hemoglobin may interfere in the Alpha 2 and Beta region.

- Samples with high concentration of lipids may show altered electropherograms.

- Presence of intravenous contrast material, medication or plasma expanders that absorbs at 214 nm will result in altered profile.

19. Quality Control

Pooled control sera or commercially available control should be included in each run.

20. References

References


- Chen, F.A., et al., Capillary Electrophoresis  A new clinical Tool, Clin.
Chem., 37(1) (19910
- Kim, J.W. et al., Quantitaive Analysis of Serum Proteins Separated by
Capillary Electrophoresis, Clin.Chem., 39:689 (1993)
- Bossuyt, X., et al., Serum protein electrophoresis by CZE 2000 clinical
capillary electrophoresis system., Clin. Chem. 44:4 749 (1998).
- Bossuyt, X., Separation of Serum Proteins by Automated Capillary Zone
Electrophoresis, Clin. Chem. Lab.Med., 41:6 762 (2003)
- Jenkins M. A., Quality control and quality assurance aspects of the routine
use of capillary electrophoresis for serum and urine proteins in clinical
laboratories, Electrophoresis 25, 1555 (2004).
- Joliff, C.R. Classification and Interpretation of Paragon Serum Protein
Electrophoresis Patterns, Beckman Coulter Technical Bulletin EP1,
Beckman Coulter, Fullerton, C.A.
- Ritchie, R.F., et al., Serum Proteins in Clincal Medicine Volume I
Laboratory Section Foundation for Blood Research PO Box 190,
Scarborough Maine USA.

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