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ANALIS

HS CEofix™ - CDT kit pour Beckman Coulter P/ACE™ 5000 series - GB: Instructions for use

QUOTATION
INFORMATION
QUOTATION
INFORMATION
DESCRIPTION TECHNICAL DESCR.
DESCRIPTION

40 tests par kit

1. Date: 11 -Dec-03 - Revision: 02

2. Intended use:

The kit is intended for the determination of human Tf (transferrin isoforms) and quantitative determination of CDT (carbohydrate deficient transferrin) in human serum.


3. Principle of the test:

The Tf are separated inside a capillary under the influence of an electrical field. Differences in charge to mass ratio give different migration times.

A UV detector is used and detects the Tf at 200nm.

The kit consists of buffers and method. The buffers use the patented dynamic double coating technique.

CDT is defined as the percent of 0-sialo Tf plus 2-sialo Tf in relation to the sum of all Tf isoforms.

4. Content:

Rinse:
1x4.2mL aqua bidistallata (PN 10-004263)
Conditioner:
1x4.2mL NaOH 0.2 M (PN 10-004261)
Initiator:
1x 4.2 mL TRIS/Phosphate buffer pH 2.0
(PN 10-004264)
Buffer:
3 x 4.2 mL TRIS/ borate buffer pH 8.5 (PN 10-004262)
Fe solution:
1 x 3.2 mL (PN 10-004265)
Micro vials:
50 (PN 10-004246)

5. Configuration of instrument:


- BeckmanCoulter P/ACE 5000

  • UV detector
  • P/ACE station software version 1.0.

6. Other material needed:


- Vials (PN 358807)

  • caps (PN 359079)
  • spring (PN 338488)
  • Cartridge 100x800 µm ( PN 727610)
  • Pipettes
  • Capillary 50µM ID x 40 cm to the detector (PN 10-004748/844111044)


7. Training:

The operator should be familiar with instrument (P/ACE 5000) 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 is the preferred specimen.

10. Instrument preparation:

Load the instrument as follow

figure 1

Programmation de l'instrument :

1 figure 2

2 figure 3


3 figure 4


4 figure 5
figure 6


5 figure 7

figure 8


6 figure 9
figure 10


7 figure 11


8 figure 12


Or download program: CDT.MET

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

11. Sample preparation:

Sample: 75 µL Fe solution + 25 µL serum in microvial and mix.

figure 13

12. Operations:

Every day operation:

  • clean instrument and interface manifold
  • put rinse, conditioner, initiator, buffer vials-
    put waste vial - put clean and dry caps on vials
  • run cond.met
    when system was down for several days

figure 14

  • prepare samples
  • place sample in instrument
  • run "CDT.MET method

End of day:

  • remove samples

  • cap reagents vials

Placing new capillary

  • When the capillary is broken or when the capillary is not operating
    properly replace the capillary: according the instrument manual.

13. Calibration:

No calibration is required for the kit.

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

14. Analysis of results:

Click to see examples.

15. Calculation:

% CDT = sum of % 0-Sialo Tf plus % of 2-Sialo Tf.

% X-Sialo Tf = corrected area of X-Sialo Tf in relation to the sum of all Tf isoforms.

(X= 0, 2, 3, 4, 5 or 6).


16. Imprecision:

Sample

Control

Low

High

Low

High

Asialo

Disialo

Asialo

Disialo

Asialo

Disialo

Asialo

Disialo

Mean

_

0.89

0.68

3.20

_

0.94

0.75

3.59

SD

_

0.06

0.12

0.12

_

0.06

0.07

0.13

CV

_

7.2

18.7

3.9

_

6.6

9.0

3.6

NCCLS EP5-A (N=80) Lanz C., Thormann W. University of Bern Switzerland on P/ACE MDQ

17. Reference range:

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

figure 15

18. Interference:
  • Heterozygote Tf variants, such as BC Tf and DC Tf are easily detected by observing two peaks of about 40% corrected area percent.
  • Some monoclonal or biclonal gammapathie can interact in the Tf region.

19. Quality Control:

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

20. References:

1. Legros F. et al. Carbohydrate-deficient Transferrin Isoforms Measured by Capillary Zone Elelectrophoresis for Detection of Alcohol Abuse. Clin Chem 2002;48: 2177-2186
2. Lanz C. et al. Evaluation and optimization of capillary zone electrophoresis with different dynamic capillary coatings for the determination of carbohydrate-deficient transferrin in human serum. J Chromatogr A 2002; 979: 43-57
3. Legros F. et al. Use of Capillary Zone Electrophoresis for Differentiating Excessive from Moderate Alcohol Consumption. Clin Chem 2003;49:440-449
4. Lanz C, et al. Capillary zone electrophoresis with a dynamic double coating for analysis of carbohydrate-deficient transferrin in human serum. Precision performance and pattern recognition. J Chromatog A 2003;1013:131-474.
5. Ramdani B. et al.Analyte Comigrating with Trisialotransferrin during Capillary Zone Electrophoresis of Sera from Patients with Cancer. Clin Chem 2003;49:1854-1864
6. Carchon H. et al. Diagnosis of Congenital Disorders of Glycosylation by Capillary Zone Electrophoresis of Serum Transferrin. Clin. Chem 2004;50:101-111
7. Lanz C. et al. Capillary zone elctrophoresis with a dynamic double coating for analysis of carbohydrate-deficient transferrin in human serum: Impact of resolution between disialo- and trisialotransferrin on reference limites. Electrophoresis 2003;24: 4272-4281
8. Martello S. et al. Determination of carbohydrate deficient transferrin (CDT) with capillary electrophoresis: an inter laboratory comparison. Forensic Science International 2004; 141:153-157

TECHNICAL DESCR.
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