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as performed by: University of Washington Medical Center Department of Laboratory Medicine Immunology Contact: Kathleen Hutchinson M.S., M.T. (ASCP) or Mark Wener, M.D., Director listed on the title page of each write-up before using the C-Reactive Protein in Serum NHANES 2007–2008 Public Release Data Set Information This document details the Lab Protocol for testing the items listed in the following table. File Name Variable Name SAS Label C-reactive protein(mg/dL) C-Reactive Protein in Serum NHANES 2007–2008 1. SUMMARY OF TEST PRINCIof CRP, particles consisting of a polystyrene core and a hydrophilic shell are used in order sample will form an antigen-antibody

complex with the latex particles. Light scattering, measured by a nephelometric procedure after 6 min, is proportional to i.e. rheumatoid arthritis, juvenile chronic arthritis, ankylosing spondylitis and Crohn's C-reactive protein has been of increasing interest because of the current availability of 2. SAFETY PRECAUTIONS C-Reactive Protein in Serum NHANES 2007–2008 3. COMPUTERIZATION; DATA SYSTEM MANAGEMENT Each shipment of specimens received from the NHANES mobile unit arrives with a corresponding transmittal sheet and an electronic version of the shipping/resulting file. b. After the testing is completed results from the BNII are transferred to the laboratory c. QC results are tr

ansferred to an Excel file using laboratory-developed software. This d. Sample results are transferred to an Excel file using laboratory-developed software e. Data entry is checked for errors. f. The result file is transmitted electronig. Technical support for this system is provided by Westat, Rockville, MD (1-301-294-4. SPECIMEN COLLECTION, STORAGE, AND HANDLING PROCEDURES; CRITERIA FOR C-Reactive Protein in Serum NHANES 2007–2008 a. No special instructions such as fasting or special diets are required. b. Fresh or frozen human serum, heparin and EDTA plasma samples are acceptable. c. Blood should be collected aseptically and the serum separated by standard laboratory d. The reque

sted sample volume for the assay is 1.0 mL, and the minimum sample e. Specimens may be stored in glass or plastif. Contamination or introduced particulate matter can lead to erroneous results. Heat g. Avoid repeated freeze/thaw cycles. 5. PROCEDURES FOR MICROSCOPIC EXAMINATIONS; CRITERIA FOR REJECTION 6. PREPARATION OF REAGENTS, CALIBRATORS (STANDARDS), CONTROLS, AND , cat. #OQIY 21 Each vial contains a solution of polystyrene particles coated with mouse monoclonal anti-CRP and preservatives: Gentamicin 6.25 mg/L, Amphotericin oC Avoid freezing. Rheumatology std SL , cat # OQKZ 13 The reagent is ready to use and unopened it is stable until the manufacturer's oC. Avoid freezing. 3. N Suppl

ementary Reagent/Precipitation , cat #OUMU15 The reagent is ready to use and is stable until the manufacturer's expiration date. oC. Avoid freezing. N Diluent , cat # OUMT 65 The reagent is ready to use and is stable until the manufacturer's expiration date. C-Reactive Protein in Serum NHANES 2007–2008 5. N Rx buffer 1. Dade Behring Nephelometer II Analyzer 25 wavelength analyzer and terminal equipped with a Power Macintosh 7200/75 computer (Dade oC) transfer arms; reagent, sample, standards rack stations (with barcode reading); buffer compartment; dilution racks; temperature oC) cuvette rotor; cuvette washing device; bar code wand reader; and optical system. The CRP assay par

ameter settings for the BNII instrument are as follows: Parameter Protein Name CRP Sample Dilution* 1:100 N Supplemental reagent 5 uL Latex reagent 40 uL Buffer for Reagent 60 uL N Diluent No. of Standard Points 6 Standard Dilutions 1:80-1:2560 Standard Curve Measuring Range (At initial dilution; approximate values, range is dependent upon standard value) 2. BNII dilution wells, Cat # OVIC11 3. Air Driven Ultracentrifuge, model 340400 (Beckman Instruments, Fullerton, CA). 4. Computers (Dell Computer Corporation, Round Rock, Texas). 5. Pipettors and disposable tips (Rainin, Emeryville, CA). 6. Gloves, disposable (Any manufacturer). d. Standards/Calibration Preparation Preparation of Qu

ality Control Materials C-Reactive Protein in Serum NHANES 2007–2008 7. CALIBRATION AND CALIBRATION VERIFICATION PROCEDURES a. Calibration Curve Standard/Reagent Dilution Conc[mg/dl] Prevalue [Bit] Start value [bits] Value Deviation 183847 RH Standard SL 1 1:2560.0 0.0007 2515 153 -0.99 1153 90 Suppl. Reagent “P” 2 1:1280.0 0.0015 2507 318 1.68 1675 85 CardioPhase hsCRP 3 1:640.0 0.0029 2525 702 0.22 1:320.0 0.0058 2517 1574 -2.76 1:160.0 0.0117 2544 3865 3.77 1:80.0 0.0234 2487 7589 -1.96 direct relationship of measured light scatter to concentration of C-reactive protein in 1. Three levels of control are run for each test series. If, with

in a testing series, C-Reactive Protein in Serum NHANES 2007–2008 8. PROCEDURE OPERATING INSTRUCTION1. Bring all controls and patient specimens to room temperature before use. Mix 2. Check the levels of buffer and diluent. Verify that the dilution wells are empty. 3. Turn power on BN2 and computer, the instrument will automatically initialize and b. Instrument Operation (see operator'1. Gently mix, uncap and load specimens into serum racks, with the barcode in the 2. Uncap and load reagents in the reagent racks, with the barcode in the open slot. 3. Select the CRP NHNS test for all specimens. Testing is done in singlicate. The instrument will automatically rerun the specimen with

a higher or lower dilution if 4. The instrument automatically calculates all results. After testing is completed, 5. Remove specimens, controls, and reagents. Return controls and reagents to the 6. Perform scheduled instrument maintenance (daily, weekly, monthly) as outlined 7. Follow the on-screen instructions for shutting the instrument down. c. Recording of Data 1 Using a lab developed program, specimen results are transferred from the 2. Control results are entered to the Assad. Replacement and Periodic Maintenance of Key Components 1. Daily Maintenance: C-Reactive Protein in Serum NHANES 2007–2008 2. Monthly Maintenance: 3. Semi-annual Maintenance 4. Periodic Maintenance to be p

erformed by the manufacturer's service engineer. 9. REPORTABLE RANGE OF TEST RESULTS approximately 0.02 mg/dL. This will vary slightly with different calibrator lots. The assay 10. QUALITY CONTROL (QC) PROCEDURES a. The method described in this protocol has been used for several years. The method b. Bench quality controls are used in this analytical method. Bench quality control c. The bench controls are purchased in sufficient quantity to provide serum samples for d. Bench quality controls are placed at the beginning of each analytical run. After analysis, the long-term quality control charts (Levey-Jennings) for each control material are consulted to determine if the system is in contro

l. The Levey Jennings chart plots the quality control material observations on the y-axis and the date of the C-Reactive Protein in Serum NHANES 2007–2008 11. REMEDIAL ACTION IF CALIBRATIONinvestigated to determine the root of the problem before any results are released. Consult 12. LIMITATIONS OF METHOD; INTERFERING SUBSTANCES AND CONDITIONSa. The upper reportable value is virtually unlimited. The upper limit of this assayb. The lowest reportable value is approximately 0.02 mg/dL. The lower limit of this c. Avoid contamination of assay reagents and disposables by particulate matter, d. Clear serum samples are recommended for analysis. Markedly increased serum 13. REFERENCE RANGES

(NORMAL VALUES)14. CRITICAL CALL RESULTS ("PANIC VALUES") C-Reactive Protein in Serum NHANES 2007–2008 15. SPECIMEN STORAGE AND HANDLING DURING TESTING16. ALTERNATIVE METHODS FOR PERFORMING TEST OR STORING SPECIMENS IF There are no acceptable alternative methods of analysis. Specimens may be stored at 4-17. TEST RESULT REPORTING SYSTEM; PROTOCOL FOR REPORTING CRITICAL 18. TRANSFER OR REFERRAL OF SPECIMENS; PROCEDURES FOR SPECIMEN the NHANES Repository in Rockville, MD for long-term storage. C-Reactive Protein in Serum NHANES 2007–2008 SUMMARY STATISTICS AND QC GRAPHS 2007-2008 C-Reactive Protein Quality Control1/23/20075/3/20078/11/200711/19/20072/27/20086/6/20089/14/2008

12/23/2008 522635226110/01 C-Reactive Protein in Serum NHANES 2007–2008 1. Wener MH, Daum PR, McQuillin, GM. The influence of age, sex, and race on the upper 2. Pearson TA, Mensah GA, Alexander RW, 3. Pepys MB. C-reactive protein fifty years on. Lancet 1981;1:653-7. 4. Sölter J, undUhlenbruck G. NeueAspekte des c-reaktiven proteins. Dtsch Med Wschr 5. Ziegenhagen G, Drahovsky D. Klinische bedeutung des c-reaktiven proteins. Med Klin 6. Dixon JS, Bird HA, Sitton NG, Pickup ME, Wri7. Kind CRH, Pepys MB. The role of serum C-reactive protein(CRP) measurement in 8. Hanson LA, Wadsworth CH. Das c-reaktive protein und sein diagnostischer wert, 9. Gerwurz H, Mold C, Siegel J, Fiedel