synchronsystem(s) c4 chemistryinformationsheet

SYNCHRON System(s) C4Chemistry Information Sheet© 2015 Beckman Coulter, Inc. All rights reserved.

Complement C4988471

For In Vitro Diagnostic Use

Rx Only

ANNUAL REVIEW

Reviewed by Date Reviewed by Date

PRINCIPLEINTENDED USE

C4 reagent, when used in conjunction with UniCel® DxC 600/800 System(s) and SYNCHRON® Systems CAL 1, isintended for quantitative determination of complement c4 concentration in human serum or plasma by turbidimetry.

CLINICAL SIGNIFICANCE

Complement is a group of serum proteins which destroy infectious agents. Measurements of these proteins aid in thediagnosis of immunologic disorders, especially those associated with deciencies of complement components.1,2

METHODOLOGY

C4 reagent is used to measure the analyte concentration by a turbidimetric method.3 In the reaction, complement c4combines with specic antibody to form insoluble antigen-antibody complexes.

The SYNCHRON System(s) automatically dilutes sample and dispenses the appropiate sample and reagent volumesinto a cuvette. The ratio used is one part diluted sample to 5.85 parts reagent. The system monitors the change inabsorbance at 340 nanometers. This change in absorbance is proportional to the concentration of complement c4 in thesample and is used by the SYNCHRON System(s) to calculate and express the complement c4 concentration basedupon a single-point calibration curve.

CHEMICAL REACTION SCHEME

Chemistry Information Sheet A18470 AN EN C4DECEMBER 2015 of 12

SPECIMENTYPE OF SPECIMEN

Serum samples are recommended. Plasma samples (Lithium Heparin, and Sodium Heparin) can be used.

Serum or plasma samples should be collected in the manner routinely used for any clinical laboratory test.4 Freshlydrawn serum or plasma from a fasting individual is preferred. Anticoagulants tested are listed in the PROCEDURALNOTES section of this chemistry information sheet.

SPECIMEN STORAGE AND STABILITY

1. Tubes of blood are to be kept closed at all times and in a vertical position. It is recommended that the serum orplasma be physically separated from contact with cells within two hours from the time of collection.5

2. Separated serum or plasma should not remain at room temperature longer than 6 hours, and no longer than 24hours at +2°C to +8°C. Specimens can be stored frozen at -15°C to -20°C for one week. Analyte deterioration mayoccur in samples that are repeatedly frozen and thawed.5

Additional specimen storage and stability conditions as designated by this laboratory:

SAMPLE VOLUME

The optimum volume, when using a 0.5 mL sample cup, is 0.3 mL of sample. For optimum primary sample tube volumesand minimum volumes, refer to the Primary Tube Sample Template for your system.

CRITERIA FOR UNACCEPTABLE SPECIMENS

Refer to the PROCEDURAL NOTES section of this chemistry information sheet for information on unacceptablespecimens.

Criteria for sample rejection as designated by this laboratory:

PATIENT PREPARATION

Special instructions for patient preparation as designated by this laboratory:

C4 EN Chemistry Information Sheet A18470 AN of 12 DECEMBER 2015

SPECIMEN HANDLING

Special instructions for specimen handling as designated by this laboratory:

REAGENTSCONTENTS

Each kit contains the following items:

Two Complement C4 Reagent Cartridges (2 x 100 tests)One lot-specic Parameter Card

VOLUMES PER TEST

Sample Dilution Volumes

Sample Volume 15 µL

Diluent Volume 285 µL

Diluted Sample Volume (1:20 dilution) 40 µL

Total Reagent Volume 234 µL

Cartridge Volumes

A 200 µL

B 34 µL

C – –

REACTIVE INGREDIENTS

REAGENT CONSTITUENTSCompartment A

Reaction Buffer 35.0 mL

Compartment B

Antibody (polyclonal) Monospecic for complementc4 (Goat)

6.0 mL

Sodium Azide (used as a preservative) < 0.1% (w/w)

Also non-reactive chemicals necessary for optimal system performance.


CAUTION

Sodium azide preservative may form explosive compounds in metal drain lines.See NIOSH Bulletin: Explosive Azide Hazard (8/16/76).To avoid the possible build-up of azide compounds, ush wastepipes withwater after the disposal of undiluted reagent. Sodium azide disposal must be inaccordance with appropriate local regulations.

CAUTION

Although not composed of substances of human origin, this product may comein contact with human serum during processing. This material and all patientsamples should be handled as though capable of transmitting infectious disease.The United States Food and Drug Administration recommends such samplesbe handled as specied in the Centers for Disease Control‘s Biosafety Level 2guidelines.6

GHS HAZARD CLASSIFICATION

C4 Reagent (Compartment A) WARNING

H319 Causes serious eye irritation.

H412 Harmful to aquatic life with long lasting effects.

P273 Avoid release to the environment.

P280 Wear protective gloves, protective clothing and eye/faceprotection.

P305+P351+P338 IF IN EYES: Rinse cautiously with water for several minutes.Remove contact lenses, if present and easy to do. Continuerinsing.

P337+P313 If eye irritation persists: Get medical advice/attention.

Polyoxyethylated Octyl Phenol < 3%

Safety Data Sheet is available at techdocs.beckmancoulter.com.

EUROPEAN HAZARD CLASSIFICATION

C4 Reagent (Compartment A) R52/53

R52/53 Harmful to aquatic organisms, may cause long-termadverse effects in the aquatic environment.

S61 Avoid release to the environment. Refer to specialinstructions/Safety data sheets.

MATERIALS NEEDED BUT NOT SUPPLIED WITH REAGENT KIT

SYNCHRON® Systems CAL 1DIL 1 Cartridge


At least two levels of control materialSaline

REAGENT PREPARATION

No preparation is required.

ACCEPTABLE REAGENT PERFORMANCE

The acceptability of a reagent is determined by successful calibration and by ensuring that quality control results arewithin your facility’s acceptance criteria.

REAGENT STORAGE AND STABILITY

C4 reagent, when stored unopened at +2°C to +8°C, will obtain the shelf-life indicated on the cartridge label. Onceopened, the reagent is stable for 60 days at +2°C to +8°C unless the expiration date is exceeded. DO NOT FREEZE.

DIL 1 stored unopened at room temperature is stable until the expiration date indicated on each cartridge. Once opened,DIL 1 is stable for 60 days on instrument or until the expiration date, if sooner.

Reagent storage location:

CALIBRATIONCALIBRATOR REQUIRED

SYNCHRON® Systems CAL 1

CALIBRATOR PREPARATION

No preparation is required.

CALIBRATOR STORAGE AND STABILITY

SYNCHRON® Systems CAL 1 is stable until the expiration date printed on the calibrator bottle if stored capped in theoriginal container at +2°C to +8°C. DO NOT FREEZE.

Calibrator storage location:


CAUTION

Because this product is of human origin, it should be handled as though capableof transmitting infectious diseases. Each serum or plasma donor unit usedin the preparation of this material was tested by United States Food and DrugAdministration (FDA) approved methods and found to be negative for antibodiesto HIV and HCV and nonreactive for HbsAg. Because no test method can offercomplete assurance that HIV, hepatitis B virus, and hepatitis C virus or otherinfectious agents are absent, this material should be handled as though capableof transmitting infectious diseases. This product may also contain other humansource material for which there is no approved test. The FDA recommends suchsamples to be handled as specied in Centers for Disease Control’s BiosafetyLevel 2 guidelines.6

CALIBRATION INFORMATION

1. The system must have a lot-specic parameter card and a valid calibration adjustment in memory before controlsor patient samples can be run.

2. Under typical operating conditions the C4 reagent cartridge must be calibrated every 30 days and also with certainparts replacements or maintenance procedures, as dened in UniCel DxC 600/800 System Instructions For Use(IFU) manual. This assay has within-lot calibration available. Refer to the UniCel DxC 600/800 System InstructionsFor Use (IFU) manual for information on this feature.

3. For detailed calibration instructions, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

4. The system will automatically perform checks on the calibration and produce data at the end of calibration. In theevent of a failed calibration, the data will be printed with error codes and the system will alert the operator of thefailure. For information on error codes, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

TRACEABILITY

For Traceability information refer to the Calibrator instructions for use.

QUALITY CONTROLAt least two levels of control material should be analyzed daily. In addition, these controls should be run with each newcalibration, with each new bottle of reagent, and after specic maintenance or troubleshooting procedures as detailed inthe appropriate system manual. More frequent use of controls or the use of additional controls is left to the discretion ofthe user based on good laboratory practices or laboratory accreditation requirements and applicable laws.

The following controls should be prepared and used in accordance with the package inserts. Discrepant quality controlresults should be evaluated by your facility.

Table 1.0 Quality Control Material

CONTROL NAME SAMPLE TYPE STORAGE


Table 1.0 Quality Control Material, ContinuedCONTROL NAME SAMPLE TYPE STORAGE

TESTING PROCEDURE(S)1. If necessary, load the reagent onto the system. A lot-specic parameter card must be loaded one time for each lot.

2. After reagent load is completed, calibration is required.

3. Program samples and controls for analysis.

4. After loading samples and controls onto the system, follow the protocols for system operations.

For detailed testing procedures, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

CALCULATIONSThe SYNCHRON System(s) performs all calculations internally to produce the nal reported result. The system willcalculate the nal result for sample dilutions made by the operator when the dilution factor is entered into the systemduring sample programming.

REPORTING RESULTSEquivalency between the SYNCHRON LX and UniCel DxC 600/800 Systems has been established. Chemistry resultsbetween these systems are in agreement and data from representative systems may be shown.

REFERENCE INTERVALS

Each laboratory should establish its own reference intervals based upon its patient population. The reference intervalvalues for C4 were established using the Synchron® System(s) C4 assay for a population of 130 apparently healthy,male and female adults from Southern California. The interval was veried with the SYNCHRON System(s) C4 assayfor an additional 125 samples.

Table 2.0 Reference intervals

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSBeckman Coulter Serum or Plasma (Adult) 18 – 55 mg/dL 180 – 550 mg/L

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSLaboratory

Refer to References (7,8,9) for guidelines on establishing laboratory-specic reference intervals.

Additional reporting information as designated by this laboratory:


PROCEDURAL NOTESANTICOAGULANT TEST RESULTS

The following anticoagulants were assessed by Deming regression analysis with a minimum of 130 paired serum andplasma samples. Values of serum (X) ranging from 13.8 mg/dL to 49.8 mg/dL were compared with the values for plasma(Y) yielding the following results:

Table 3.0 Anticoagulant Test Results

ANTICOAGULANT LEVEL OF ANTICOAGULANT TESTEDDEMING REGRESSION ANALYSIS

(mg/dL)Lithium Heparin 14 Units/mL Y= 0.899X + 1.21; r = 0.978

Sodium Heparin 14 Units/mL Y = 0.900X + 0.76; r = 0.992

LIMITATIONS

Activation of C4 can occur both in vivo and in vitro and results in the production of two breakdown products; C4a andC4b. The antibody employed in the Beckman Coulter C4 assay is directed against the common portion of the moleculeand will exhibit the same reactivity with C4 fragments as it does with the native molecule.

INTERFERENCES

1. The following substances were tested in serum for interference with this methodology:

Table 4.0 Interferences

SUBSTANCE SOURCE LEVEL TESTED OBSERVED EFFECTBilirubin Porcine 30 mg/dL NSIa

Lipemia Human 3+ NSI

Hemoglobin Human 500 mg/dL NSI

Rheumatoid Factor Human 300 IU/mL NSI

Paraprotein (IgM) Human 500 mg/dL NSIa NSI = No Signicant Interference (within ±4.0 mg/dL or 9.0%).

2. For lipemic specimens with visual turbidity level >3+ or serum index >8, ultra-centrifugation (90,000 x g for 10minutes) is recommended for accurate C4 results.

3. Refer to References (10,11,12) for other interferences caused by drugs, disease and preanalytical variables.

PERFORMANCE CHARACTERISTICSANALYTIC RANGE

The SYNCHRON System(s) method for the determination of this analyte provides the following analytical range:

Table 5.0 Analytical Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSSerum or Plasma 5.0 – 120.0 mg/dL 50 – 1200 mg/L

Samples with concentrations outside the analytical range will be reported as "<5 mg/dL" ("<50 mg/L") or ">120 mg/dL"(">1200 mg/L").


Samples reported out as greater than the analytical range should be conrmed by diluting with saline and reanalyzing.If a manual dilution is used the appropriate dilution factor should be applied to the reported result.

REPORTABLE RANGE (AS DETERMINED ON SITE):

Table 6.0 Reportable Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS

SENSITIVITY

Sensitivity is dened as the lowest measurable concentration which can be distinguished from zero with 95% condence.Sensitivity for C4 determination is 5 mg/dL (50 mg/L).

EQUIVALENCY

Equivalency was assessed by Deming regression analysis of freshly drawn patient samples to Beckman Coulter’sIMMAGE nephelometric immunoassay system. Updated IMMAGE comparison data is provided for crossoverinformation only. The Synchron® Systems Calibrator 1 set point for C4 has been value-assigned to produce C4assay recovery to match the ERM-DA470k target value. Please follow the proper guidance or established laboratoryrequirements for crossover testing.

Serum (in the range of 7.1 to 101.1 mg/dL)Y (SYNCHRON DxC Systems) = 1.558X - 5.09

N = 100

MEAN (SYNCHRON DxC Systems) = 35.1

MEAN (Nephelometric Immunoassay) = 25.8

CORRELATION COEFFICIENT (r) = 0.996

Refer to References (8,13) for guidelines on performing equivalency testing.

PRECISION

A properly operating SYNCHRON System(s) should exhibit precision values less than or equal to the following:

Table 7.0 Precision Values

1 SD CHANGEOVER VALUEaTYPE OFPRECISION SAMPLE TYPE mg/dL mg/L mg/dL mg/L % CV

Within-run Serum or Plasma 2.0 20.0 44.4 444 4.50

Total Serum or Plasma 3.0 30.0 44.4 444 6.75a When the mean of the test precision data is less than or equal to the changeover value, compare the test SD to the SD guideline given above to

determine the acceptability of the precision testing. When the mean of the test precision data is greater than the changeover value, compare thetest % CV to the guideline given above to determine acceptability. Changeover value = (SD guideline/CV guideline) x 100.

Comparative performance data for a SYNCHRON LX® System evaluated using the NCCLS Proposed Guideline EP5-T2appears in the table below.14 Each laboratory should characterize their own instrument performance for comparisonpurposes.


Table 8.0 NCCLS EP5-T2 Precision Estimate Method

EP5-T2 CalculatedPoint EstimatesTYPE OF

IMPRECISION SAMPLE TYPENo.

SystemsNo. DataPointsa

Test MeanValue(mg/dL) SD % CV

Serum Control 1 1 80 25.1 0.40 1.59

Serum Control 2 1 80 38.0 0.50 1.32

Serum Control 3 1 80 50.1 0.85 1.70

Serum Pool 1 1 80 9.5 0.23 2.37

Within-run

Serum Pool 2 1 80 70.3 1.07 1.53

Serum Control 1 1 80 25.1 0.46 1.82

Serum Control 2 1 80 38.0 0.64 1.68

Serum Control 3 1 80 50.1 1.09 2.17

Serum Pool 1 1 80 9.5 0.29 3.09

Total

Serum Pool 2 1 80 70.3 1.34 1.91a The point estimate is based on the data from one system, run for twenty days, two runs per day, two observations per run on an instrument

operated and maintained according to the manufacturer‘s instructions.

Refer to References (8,15) for guidelines on performing precision testing.

NOTICE

These degrees of precision and equivalency were obtained in typical testing procedureson a SYNCHRON LX® System and are not intended to represent the performancespecications for this reagent.

ADDITIONAL INFORMATIONFor more detailed information on UniCel DxC Systems, refer to the appropriate system manual.

Beckman Coulter, the Beckman Coulter Logo, Synchron, UniCel and DxC are trademarks of Beckman Coulter, Inc andare registered in the USPTO.

SHIPPING DAMAGE

If damaged product is received, notify your Beckman Coulter Clinical Support Center.

REVISION HISTORY

Revision AF

Updated corporate address; updated European Hazard Classication and removed EDTA as an AcceptableAnticoagulant claim.

Revision AG

Removed EDTA recommendation claim from Type of Specimen section.

Revision AH

Added Revision History.


Revision AJ

Revised the Reference Range and Equivalency section.

Revision AK

Added new language requirement: Czech, and Korean.

Revision AL

Removed references to CX and LX systems as they are discontinued effective 12/2013.

Added Beckman Coulter trademark statement and disclaimer.

Revision AM

Added GHS Classication information

Revision AN

Added new language requirement: Romanian


REFERENCES1. Nusinow, SR, Zuraw, BL and Curd, JG, "The Hereditary and Acquired Deficiencies of Complement", Medical Clinics

of North America, (1981).

2. Ross, SC and Densen, P, "Complement Deficiency States and Infection: Epidemiology, Pathogenesis andConsequences of Neisserial and Other infections in an Immune Deficiency", Medicine, (1984).

3. Whicher, J. T., Price, C. P., Spencer, K., "Immunonephelometric and Immunoturbidimetric Assays for Proteins",Crit. Rev., Clin. Lab. Sci, 18:213 260 (1983).

4. Tietz, N. W., "Specimen Collection and Processing; Sources of Biological Variation", Textbook of ClinicalChemistry, 5th Edition, W. B. Saunders, Philadelphia, PA (2005).

5. National Committee for Clinical Laboratory Standards, Procedures for the Handling and Processing of BloodSpecimens Approved Guideline, NCCLS publication H18-A, Villanova, PA (1990).

6. CDC-NIH, Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, (Washington, D.C.: U.S.Government Printing Office, 2009). (CDC 21-1112)

7. National Committee for Clinical Laboratory Standards, How to Define, Determine, and Utilize Reference Intervalsin the Clinical Laboratory Approved Guideline, NCCLS publication C28-A, Villanova, PA (1995).

8. Burtis, C. A., Ashwood, E. R., Tietz, Textbook of Clinical Chemistry, 3rd Edition, W. B. Saunders, Philadelphia, PA(1999).

9. Henry, J. B., Clinical Diagnosis and Management by Laboratory Methods, 22nd Edition, W. B. Saunders Company,Philadelphia, PA (2006).

10. Young, D. S., Effects of Drugs on Clinical Laboratory Tests, 5th Edition, AACC Press, Washington, D. C. (2000).

11. Friedman, R. B., Young, D. S.,Effects of Disease on Clinical Laboratory Tests, 4th Edition, AACC Press,Washington, D.C. (2001).

12. Young, D. S., Effects of Preanalytical Variables on Clinical Laboratory Tests, 3rd Edition, AACC Press,Washington, D. C. (2007).

13. National Committee for Clinical Laboratory Standards, Method Comparison and Bias Estimation Using PatientSamples Approved Guideline, NCCLS publication EP9-A, Villanova, PA (1995).

14. National Committee for Clinical Laboratory Standards, Precision Performance of Clinical Chemistry DevicesTentative Guideline, 2nd Edition, NCCLS publication EP5-T2, Villanova, PA (1992).

15. National Committee for Clinical Laboratory Standards, Precision Performance of Clinical Chemistry Devices, 2ndEdition, Approved Guideline, NCCLS publication EP5-A, Wayne, PA (1999).

Beckman Coulter Eurocenter S.A., 22, rue Juste-Olivier. Case Postale 1044, CH - 1260 Nyon 1, SwitzerlandTel: +41 (0)22 365 36 11

Beckman Coulter, Inc., 250 S. Kraemer Blvd., Brea, CA 92821 U.S.A.


synchronsystem(s) c4 chemistryinformationsheet

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