This Procedure Is Valid for the Following Chemistry Analyzers s6

Procedure: PREALBUMIN OSR6175 This procedure is valid for the following chemistry analyzers:

 AU400/AU400e  AU640/AU640e  AU480  AU680  AU600  AU2700  AU5400  AU5800

Prepared By Date Adopted Supersedes Procedure #

Review Date Revision Date Signature

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Prealbumin is a transport protein. It plays a significant role in the metabolism of vitamin A by complexing with retinol-binding protein, which in turn complexes with Vitamin A transporting it through the body. This protein is an acute phase reactant whose levels fall in inflammation, malignancy, cirrhosis of the liver and protein wasting diseases of the gut or kidneys. Decreases are associated with liver disease and malnutrition. Levels of Prealbumin increase in Hodgkins disease.1,2

INTENDED USE:

System reagent for the quantitative determination of Prealbumin in human serum on Beckman Coulter AU Clinical Chemistry analyzers.

METHODOLOGY:

Immune complexes formed in solution scatter light in proportion to their size, shape and concentration. Turbidimeters measure the reduction of incident light due to reflection, absorption, or scatter.

In the Beckman Coulter AU procedure, the measurement of the decrease in light transmitted (increase in absorbance) through particles suspended in solution as a result of complexes formed during the antigen-antibody reaction is the basis of this assay.

PATIENT PREPARATION:

An 8 to 12-hour fast is recommended, but not required, in order to minimize any possible lipemia.

Additional instructions for patient preparation as designated by this laboratory:

Fasting serum samples, free from hemolysis are the recommended specimens. Avoid highly lipemic samples, which may produce excessively high scatter signals.3 Samples containing Intralipid* may cause interference in this assay. *Intralipid, manufactured by KabiVitrium Inc., is a 20% IV fat emulsion.

Additional type conditions as designated by this laboratory:

HANDLING CONDITIONS:

Serum samples are stable 7 days when stored at 2 - 8C. These samples can be stored frozen (<-20ºC) for 2 months.

Additional handling conditions as designated by this laboratory:

EQUIPMENT AND MATERIALS:

EQUIPMENT:

Beckman Coulter AU400/AU400e, AU480, AU600, AU640/AU640e, AU680, AU2700, AU5400 and AU5800 analyzers.

MATERIALS:

Beckman Coulter AU System Prealbumin Reagent

Final concentration of reactive ingredients: Solution of Polymers in Phosphate Buffered Saline (pH 7.1 - 7.3) Rabbit anti-human Prealbumin antiserum

Also contains preservatives.

Reagent storage location in this laboratory:

Test tubes 12 -16 mm in diameter or sample cups (Cat No. AU1063).

Storage location of test tubes or sample cups in this laboratory:

Beckman Coulter Prealbumin Calibrator (Cat No. ODR3029)

Storage location of the calibrator in this laboratory:

Precautions: 1. For in vitro diagnostic use.

2. Do not ingest reagent or calibrator. Harmful if swallowed.

3. Contains sodium azide as a preservative that may react with lead joints in copper plumbing to form explosive compounds. Even though the reagent contains minute quantities of sodium azide, drains should be well flushed with water when discarding the reagent or calibrators.

4. WARNING: POTENTIAL BIOHAZARDOUS MATERIAL. The calibrator is manufactured from human serum. No test method can offer complete assurance that HIV- 1/2, HCV, Hepatitis B, or other infectious agents are absent from biological materials, all calibrator material should be handled at the Biosafety Level 2 as recommended for any infectious human serum or blood specimen in the CDC/National Institutes of Health manual, Biosafety in Microbiological and Biomedical Laboratories, 1993.

PREPARATION:

The Beckman Coulter AU System Prealbumin reagent is ready for use. No preparation is required.

The Beckman Coulter Prealbumin Calibrators are liquid, ready for use as supplied. Mix by gentle inversion to achieve a homogenous mixture prior to use.

Storage Requirements: 1. The unopened reagents and calibrators are stable until the expiration date printed on the label when stored at 2 - 8C.

2. Opened bottles of reagent are stable for 90 days when stored in the refrigerated compartment of the Beckman Coulter AU analyzers.

© Beckman Coulter, Inc. March 2012 CLSIOSR6x75.02 All printed copies are considered to be copies of the electronic original. Page 5 of 11 Procedure: PREALBUMIN OSR6175 3. Opened bottles of calibrator are stable until the expiration date printed on the label providing that the stoppers and caps are replaced immediately after each use. The Prealbumin calibrators should be stored at 2 – 8C when not in use.

Additional storage requirements as designated by this laboratory.

Indications of Deterioration: Discoloration of the reagent or calibrators, visible signs of microbial growth, turbidity or precipitation in reagent or calibrators may indicate degradation and warrant discontinuance of use.

PERFORMANCE PARAMETERS:

The following data was obtained using this Prealbumin Reagent on Beckman Coulter AU analyzers according to established procedures. Results obtained at individual laboratories may differ.

PRECISION:8

Estimates of precision, based on CLSI recommendations7, are consistent with typical performance. The within run precision is less than 5%CV and total precision is less than 10% CV. Assays of serum pools and control sera were performed and the data reduced following CLSI guidelines.

N=100 Within run Total Mean, mg/dL SD CV% SD CV% 14.1 0.09 0.6 0.20 1.5

25.3 0.14 0.6 0.29 1.1 35.5 0.35 1.0 0.42 1.2

COMPARISON: 8

Patient samples were used to compare the Beckman Coulter AU System Prealbumin Reagent. Representative performance data on AU analyzers is shown in the next table. Y Method AU640®/AU640e® X Method AU600® Slope 1.028 Intercept -0.66 Correlation Coeff. (r) 0.999 No. of Samples (n) 158 Range (mg/dL) 5 - 57

CALIBRATION:

STANDARD PREPARATION:

Perform a multi-point calibration (5AB) using a water blank (blue rack) and the appropriate calibrators in a yellow calibration rack. The frequency of calibration is every 90 days. Calibration of this Prealbumin procedure is accomplished by use of the Beckman Coulter AU Prealbumin Calibrator (Cat No. ODR3029) which is traceable to IFCC International Reference Preparation CRM470 (RPPHS).

This calibrator has not been tested for use with any other Chemistry Systems other than those listed above.

The results obtained using this calibrator are dependent upon several factors, including proper storage of the calibrator and proper technique in use of the Beckman Coulter AU Clinical Chemistry analyzers and their respective reagents.

Recalibration is required when any of the following conditions occur:

1. A reagent lot number has changed or there is an observed shift in control values.

2. Major preventative maintenance was performed on the analyzer.

3. A critical part was replaced.

QUALITY CONTROL:

During operation of the Beckman Coulter AU analyzer at least two levels of an appropriate immunology control material should be tested a minimum of once a day. In addition, these controls should be performed after calibration, with each new lot of reagents, and after specific maintenance or troubleshooting steps described in the appropriate AU User’s Guide. Quality control testing should be performed in accordance with regulatory requirements and each laboratory’s standard procedure.

Location of controls used at this laboratory.

ANALYZER PARAMETERS:

A complete list of test parameters and operating procedures can be found in the appropriate User’s Guide and at www.beckmancoulter.com.

CALCULATIONS:

For SI units (g/L), multiply the results by 0.01.

REFERENCE RANGES:

Adults6: 17 - 34 mg/dL

This reference range was determined from healthy blood donors (data on file). Expected values may vary with age, sex, diet and geographical location. Each laboratory should determine its own expected values as dictated by good laboratory practice.

Expected reference ranges in this laboratory:

PROCEDURES FOR ABNORMAL RESULTS:

Abnormal results are flagged by the listed analyzers according to the normal values entered by the user into the instrument parameters.

REPORTING FORMAT:

Results are automatically printed out for each sample in mg/dL at 37o C.

Additional reporting information as designated by this laboratory:

The Beckman Coulter AU System Prealbumin assay is linear from 3 - 80 mg/dL. Samples exceeding the upper limit of linearity should be diluted and repeated. The sample may be diluted, repeated and multiplied by the dilution factor automatically utilizing the AUTO REPEAT RUN.

INTERFERING SUBSTANCES:

Results of studies4 conducted show that the following substances may interfere with this prealbumin procedure:

The criteria for no significant interference is recovery within 10% of the initial value. Ascorbate: No significant interference up to 20 mg/dL Ascorbate Bilirubin: No significant interference up to 40 mg/dL Bilirubin Hemolysis: No significant interference up to 500 mg/dL Hemolysate Lipemia: No significant interference up to 225 mg/dL Triglyceride (Avian)**

**Measured at prealbumin concentrations of 10 & 40 (+/-5) mg/dL.

The information presented is based on results from Beckman Coulter AU studies and is current at the date of publication. Beckman Coulter Inc., makes no representation about the completeness or accuracy of results generated by future studies. For further information on interfering substances, refer to Young5 for a compilation of reported interferences with this test.

Laboratory specific procedure notes:

1. Tietz N.W., Textbook of Clinical Chemistry, Second Edition, Burtis, C.A., and Ashwood E.R., editors, W.B. Saunders Company, 1994.

2. Kaplan L.A. and Pesce A.J., Clinical Chemistry Theory Analysis, Correlation, Third Edition, Mosby, St. Louis, Missouri, 1996.

3. Rose, N.R., Friedman, H, and Fahey, J.L., Manual of Clinical Laboratory Immunology, Third Edition, American Society for Microbiology, Washington, DC, 1986.

4. CLSI/NCCLS, Interference Test in Clinical Chemistry, EP7-P, 1986.

5. Young, D.S., Effects of Drugs on Clinical Laboratory Tests, AACC Press, Fifth Edition, 2000.

6. Beckman Coulter Inc. data on samples collected from 200 blood donors in North Texas.

7. CLSI/NCCLS Evaluation Protocol EP5-T2, 1992.

8. Data is on file for specific AU analyzers.