RANDOX LIQUID CK-MB, RANDOX CK-MB CALIBRATOR

{0} 1 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k123977 B. Purpose for Submission: New device C. Measurand: Creatine kinase isoform MB (CKMB) D. Type of Test: Quantitative immunoinhibition, kinetic rate immunoassay E. Applicant: Randox Laboratories Limited F. Proprietary and Established Names: Randox Liquid CK-MB, Randox CK-MB Calibrator G. Regulatory Information: 1. Regulation section: 21CFR 862.1215 21 CFR 862.1150 2. Classification: Class II 3. Product code: CGS, JIT 4. Panel: Chemistry 75 {1} H. Intended Use: 1. Intended use(s): Randox Liquid CK-MB Please see below. 2. Indication(s) for use: Randox Liquid CK-MB: The Randox Liquid CK-MB test system is a device intended for the quantitative in vitro determination of CK-MB concentration in serum and plasma. Measurements of CK-MB are used in the diagnosis and treatment of myocardial infarction (MI). This product is suitable for use on the RX series instruments including the RX Daytona and the RX Imola. Randox CK-MB Calibrator: The Randox CK-MB calibrator is an in vitro diagnostic product intended for use in the calibration of Randox CK-MB methods. The Randox Liquid CK-MB test system for the RX Imola and RX Daytona is a prescription use device intended to be used in clinical laboratories only. 3. Special conditions for use statement(s): For prescription use. For clinical laboratory use only. 4. Special instrument requirements: For use with RX Daytona and RX Imola I. Device Description: The Randox Liquid CK-MB is supplied in a kit containing: 4 x 20.0 mL CK-MB Buffer 4 x 6.0 mL CK-MB Substrate. The Randox CK-MB calibrator is lyophilized, single analyte, human serum based product. The kit contains ten vials (single level) with 1.0 mL per vial. Double deionized water is required for reconstitution. {2} Human source material from which this product has been derived has been tested at donor level using FDA approved methods for Human Immunodeficiency Virus (HIV1, HIV2) antibody, the Hepatitis B surface antigen (HbSAg) and Hepatitis C Virus (HCV) antibody and was found to be non-reactive. However, since no method can offer complete assurance as to the absence of infectious agents, this material and all patient samples should be handled as though capable of transmitting disease and disposed of accordingly. ## J. Substantial Equivalence Information: 1. Predicate device name(s): Roche Diagnostics CK-MB reagent, Roche Diagnostics CK-MB calibrator for automated systems 2. Predicate 510(k) number(s): k003158 3. Comparison with predicate: | CHARACTERISTICS | RANDOX LIQUID CK-MB | ROCHE DIAGNOSTIC SYSTEMS Inc. CK-MB REAGENT K003158 | | --- | --- | --- | | Intended Use | Intended for the quantitative in vitro determination of CK-MB concentration in serum and plasma. | Same | | Assay Protocol | Immunoinhibition Assay | UV assay with immunological inhibition of CK-M | | Format | Liquid which are ready to use | Same | | Antibody | Mouse anti-human CK-M monoclonal antibody | Same | | Sample Type | Plasma and serum | Plasma and serum | | Sample Tubes | Heparin, EDTA | Same | | Control | Randox CK-MB Control | Precinorm CK-MB Control | {3} | (Frequency) | Randox Tri-level Cardiac Controls levels 2 and 3 Two levels of control should be assayed at least once a day | - Control interval per institution requirements | | --- | --- | --- | | Measuring Range | Daytona: 7-2000 U/L Imola: 6-1100 U/L | 5-2300 U/L | | CHARACTERISTICS | RANDOX CK-MB CALIBRATOR | ROCHE DIAGNOSTIC SYSTEMS Inc. CK-MB CALIBRATOR K003158 | | --- | --- | --- | | Intended Use | In vitro diagnostic product intended for use in the calibration of Randox CK-MB methods. | Same | | Size | 10 x 1ml | 3 x 1ml | | Analyte | CK-MB | Same | | Matrix | Human serum | BSA | | Form | Lyophilized | Same | | Levels | Single Level | Same | K. Standard/Guidance Document Referenced (if applicable): - Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline (EP5-A2) - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline (EP6-A) - Interference Testing in Clinical Chemistry; Approved Guideline (EP7-A2), - Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline (EP9-A2-IR), - Protocols for Determination of Limits of Detection and Limits of Quantification; Approved Guideline (EP17-A) - Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; {4} Approved Guideline (C28-A3c) # L. Test Principle: The Randox Liquid CK-MB is an immunoinhibition assay. The CK reagent contains an antibody to the M subunit of CK-MB which inhibits the M subunit activity. Thus, only the activity of the B subunit is measured. The activity is multiplied by 2 yielding the activity of CK-MB in the sample. The R1 reagent contains anti CK-MM antibody (mouse monoclonal) which binds the M subunit of CK in the serum sample thereby inhibiting the activity of the CK- M subunit. The CK-B activity is determined by the CK NAC method.  The rate of increase of absorbance at $340/700\mathrm{nm}$ due to the formation of NADPH is directly proportional to the activity of CK in the sample and if the activity is multiplied by a factor of two it will give the activity of CK-MB in the sample. # M. Performance Characteristics (if/when applicable): # 1. Analytical performance: # a. Precision/Reproducibility: Precision studies were performed using control material and five spiked serum pools for CKMB. Testing was conducted in duplicate for 20 days with two runs per day $(N = 80)$ using one reagent lot on the RX Daytona and RX Imola. No recalibrations were performed during the study. The results are summarized in the following tables. Precision RX Daytona | Sample | N | Expected Value | Within | Run | Between | Run | Between | Day | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV | SD | CV | SD | CV | SD | CV | | 1 | 80 | 9.90 | 1.01 | 10.2 | 0.00 | 0.0 | 0.39 | 4.0 | 1.08 | 10.9 | | 5 | 80 | 1011.88 | 17.60 | 1.7 | 15.61 | 1.5 | 17.19 | 1.7 | 29.14 | 2.9 | | 3 | 80 | 244.85 | 4.29 | 1.8 | 2.70 | 1.1 | 3.34 | 1.4 | 6.07 | 2.5 | | 4 | 80 | 437.45 | 7.49 | 1.7 | 5.16 | 1.2 | 8.02 | 1.8 | 12.13 | 2.8 | | 2 | 80 | 47.53 | 1.00 | 2.1 | 0.78 | 1.6 | 0.89 | 1.9 | 1.55 | 3.3 | | Calibrator | 80 | 189.65 | 3.20 | 1.7 | 1.78 | 0.9 | 2.96 | 1.6 | 4.71 | 2.5 | | Control | 80 | 137.45 | 2.79 | 2.0 | 3.11 | 2.3 | 1.43 | 1.0 | 4.42 | 3.2 | {5} Precision RX Imola | Sample | N | Expected Value | Within Run | | Between Run | | Between Day | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV | SD | CV | SD | CV | SD | CV | | 1 | 80 | 10.38 | 1.24 | 12.0 | 0.00 | 0.0 | 0.49 | 4.7 | 1.33 | 12.9 | | 5 | 80 | 1001.29 | 18.07 | 1.8 | 33.15 | 3.3 | 14.51 | 1.4 | 40.45 | 4.0 | | 3 | 80 | 245.62 | 4.84 | 2.0 | 3.98 | 1.6 | 5.54 | 2.3 | 8.36 | 3.4 | | 4 | 80 | 437.83 | 8.51 | 1.9 | 6.23 | 1.4 | 9.98 | 2.3 | 14.52 | 3.3 | | 2 | 80 | 47.43 | 1.53 | 3.2 | 1.42 | 3.0 | 1.12 | 2.4 | 2.37 | 5.0 | | Calibrator | 80 | 186.69 | 5.05 | 2.7 | 0.00. | 0.0 | 3.46 | 1.9 | 6.12 | 3.3 | | Control | 80 | 135.57 | 3.70 | 2.7 | 0.39 | 0.3 | 0.39 | 0.3 | 3.74 | 2.8 | b. Linearity/assay reportable range: Linearity studies were conducted following the sponsor’s internal protocol and acceptance criteria and in accordance with CLSI document EP06-A. Two pools were used to assess the linearity of the assay used. The two pools of CK-MB stripped human serum were spiked with CK-MB and diluted to obtain 11 equally distributed levels with expected values of 7.00-2200 U/L for the RX Daytona and 6.0-1100 U/L for the RX Imola. Based on the data, linearity across the measuring range for each analyzer is summarized below: | Instrument | Slope | Intercept | Sy x | r | Measuring Range | | --- | --- | --- | --- | --- | --- | | Daytona | 1.02 | -1.27 | 26.56 | 0.99 | 7-2000 U/L | | Imola | 0.97 | 3.20 | 8.17 | 1.00 | 6-1100 U/L | Recovery studies demonstrated that the measuring range can be extended up to 10,200 U/L CK-MB for the RX Daytona and Imola analyzers to within $\pm 10\%$ of the CK-MB concentration. c. Traceability, Stability, Expected values (controls, calibrators, or methods): The Randox CK-MB calibrators have in-house traceability. Calibrator value assignment and stability: Value assignment for calibrator: A mean value for new lots of the CK-MB calibrator are established by performing nested testing of the new calibrator lot against a master lot. Values for the new lot must meet the pre-determined acceptance criteria for the device. New master lots of calibrators are evaluated against the previous master lot and values must fall within the sponsor’s predetermined acceptance criteria. CK-MB calibrator shelf life stability was evaluated in real time at $2 - 8^{\circ}\mathrm{C}$ and -75 to $-90^{\circ}\mathrm{C}$. The CK-MB calibrator once reconstituted, is stable in serum for 5 days at {6} +4°C, 8 hours at +25°C and 4 weeks at -20°C when frozen once. All data and protocols were found to be acceptable to support the stability claims above. ## Reagent stability: The Randox Liquid CK-MB reagent components (CK-MB buffer and CK-MB substrate) are ready for use. Based on real time and accelerated studies proved the reagents contained are stable up to 12 months when stored unopened at +2 to +8°C and are stable for 28 days at +10°C on board the RX Daytona and RX Imola. All data and protocols were found to be acceptable to support the stability claims above. ## d. Detection limit: Limit of Blank (LoB.), a Limit of Detection (LoD.) and a Limit of Quantification (LoQ) studies were performed in accordance with CLSI guideline EP17-A .on the RX Daytona and RX Imola systems. The LoB study analyzed 60 saline samples. Determination of the LoB was based on the 95th percentile for the results. Stripped human serum, spiked with CK-MB was used to perform the LoD and LoQ studies. The LoD study analyzed 60 samples with CK-MB values ranging from 1-4 U/L. Results were analyzed according to recommendations in CLSI EP-17A. The LoQ was determined to be the lowest concentration sample across 5 runs that had a %CV of ≤ 20% and a % deviation to the spiked target of ≤ 20%. The studies yielded the following results: | | LoB | LoD | LoQ | | --- | --- | --- | --- | | Daytona | 2.90 U/L | 5.06 U/L | 7.00 U/L | | Imola | 0.87 U/L | 2.40 U/L | 6.00 U/L | The low ends of the measuring ranges for the RX Daytona and RX Imola CK-MB assay are based on the LoQ for each analyzer. ## e. Analytical specificity: Common endogenous interfering substances were evaluated in accordance with CLSI Guideline EP7-A2. Human serum samples at approximate CK-MB concentrations of 20 U/L and 400 U/L were spiked with various concentrations of interferents and evaluated in triplicate at each interferent level. Absolute bias was calculated at each concentration level as compared with non-spiked human serum. At a given concentration, a compound was defined as causing significant interference if it caused a >±10% difference when compared to the sample negative for the interfering compound. CK-MB was evaluated at 20 U/L and the 400 U/L. No interference was observed with the exception of Intralipid. Results are summarized below: | Interferent | Concentration Tested | RX daytona | RX imola | | --- | --- | --- | --- | | Hemoglobin | 1000 mg/dl | No Interference | No Interference | {7} 8 | Bilirubin (F) | 60 mg/dl | No Interference | No Interference | | --- | --- | --- | --- | | Bilirubin (C) | 60 mg/dl | No Interference | No Interference | | Intralipid | | Interferes | Interferes | | Triglycerides | 500 mg/dl | No Interference | No Interference | | Acetaminophen | 1660μmol/l | No Interference | No Interference | | Caffeine | 308μmol/l | No Interference | No Interference | | Phenytoin | 198μmol/l | No Interference | No Interference | | Salicyclic Acid | 4.34μmol/l | No Interference | No Interference | | Digoxin | 6.15μmol/l | No Interference | No Interference | | Nicotine | 6.2μmol/l | No Interference | No Interference | | Theophylline | 222μmol/l | No Interference | No Interference | | Acetyl Salicyclic Acid | 3333μmol/l | No Interference | No Interference | | Ascorbic Acid | 227μmol/l | No Interference | No Interference | | Chloramphenicol | 155μmol/l | No Interference | No Interference | | Furosemide | 181μmol/l | No Interference | No Interference | | Ibuprofen | 2425μmol/l | No Interference | No Interference | Interference was observed with Intralipid, however triglyceride did not interfere up to 500 mg/dL. Labeling specifies that only clear, non-cloudy samples should be analyzed for CK-MB on the RX Daytona and RX Imola. f. Assay cut-off: Not applicable 2. Comparison studies: a. Method comparison with predicate device: In accordance with EP09-A2, a set of 90 patient human sera (6 spiked) spanning the range of CKMB from 8.88 to 1931.75 U/L were tested on the Daytona in singlicate, and compared to Roche CK-MB kit on the Hitachi 717 analyzer. Similarly, a set of 93 serum patient samples 9 (4 spiked) spanning the range 6.64 to 992.31 U/L were tested on the Imola and Roche CK-MB kit on the Hitachi 717 analyzer. Deming Regressions are as follows: | | Slope | Intercept | Variance ratio | Sy x | | --- | --- | --- | --- | --- | | RX Daytona | 0.95 | 0.51 | 1.00 | 11.298 | | RX Imola | 0.96 | 2.32 | 1.00 | 5.54 | Linear Regressions are as follows: {8} | | Slope | Intercept | R | Syx | | --- | --- | --- | --- | --- | | RX Daytona | 0.95 | 0.59 | 0.99 | 11.296 | | RX Imola | 0.96 | 2.36 | 0.99 | 5.54 | # b. Matrix comparison: Matrix comparison studies with serum/plasma pairs for Lithium Heparin and K EDTA plasma were performed on the RX Daytona and RX Imola. 66 natural and 6 spiked samples $(n = 72)$ were analyzed on the Daytona. 67 natural and 4 spiked samples $(n = 71)$ were analyzed on the Imola. Samples were analyzed across 5 working days in singlicate. The results from each plasma type was compared to serum and analyzed by Linear and Deming Regressions. The results are summarized below: Deming Regression: | Lithium Heparin | Slope | Intercept | Variance ratio | Syx | Sample Range | | --- | --- | --- | --- | --- | --- | | RX Daytona | 1.01 | -0.23 | 1.00 | 8.304 | 9.09-1931.8 U/L | | RX Imola | 1.00 | 0.27 | 1.00 | 8.84 | 6.8-992.3 U/L | Linear Regression: | Lithium Heparin | Slope | Intercept | R | Syx | | --- | --- | --- | --- | --- | | RX Daytona | 1.01 | -0.19 | 1.00 | 8.303 | | RX Imola | 1.00 | 0.37 | 0.99 | 8.846 | Deming Regression: | K EDTA | Slope | Intercept | Variance ratio | Syx | Sample Range | | --- | --- | --- | --- | --- | --- | | RX Daytona | 1.03 | -1.99 | 1.00 | 11.2684 | 9.09-1931.9 U/L | | RX Imola | 1.01 | 0.87 | 1.00 | 4.735 | 6.64-992.3 U/L | Linear Regression: | K EDTA | Slope | Intercept | R | Syx | | --- | --- | --- | --- | --- | | RX Daytona | 1.03 | -1.92 | 0.99 | 11.2667 | | RX Imola | 1.01 | 0.90 | 1.00 | 4.725 | # 3. Clinical studies: # a. Clinical Sensitivity: Not applicable {9} b. Clinical specificity: Not applicable c. Other clinical supportive data (when a. and b. are not applicable): Not applicable 4. Clinical cut-off: Not applicable 5. Expected values/Reference range: Verification of the CK-MB reference range was performed on the RX Daytona and RX Imola. Samples were obtained from 40 apparently healthy individuals and tested in singlicate on both test systems. The data confirmed the reference range of ≤ 25 U/L for CK-MB.¹,² References: ¹ Klein G, Berger A, Bertholf R et al. Abstract: Multicenter Evaluation of Liquid Reagents for CK, CK-MB and LDH with Determination of Reference Intervals on Hitachi Systems. Clin Chem 2001; 47:Suppl. A30. ²Thomas L, Müller M, Schumann G, Weidemann G et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. J Lab Med 2005;29:301-308. N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision. 10