RAMP CK-MB ASSAY

K033747 · Response Biomedical Corp. · JHX · May 17, 2004 · Clinical Chemistry

Device Facts

• Record ID: K033747
• Device Name: RAMP CK-MB ASSAY
• Applicant: Response Biomedical Corp.
• Product Code: JHX · Clinical Chemistry
• Decision Date: May 17, 2004
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1215
• Device Class: Class 2

Indications for Use

The RAMP CK-MB Assay is a quantitative immunochromatographic test indicated for use as an in vitro diagnostic product used with the RAMP Clinical Reader to measure CK-MB levels in EDTA whole blood. Measurement of CK-MB aids in the rapid diagnosis of acute myocardial infarction (AMI). The RAMP CK-MB Assay is not intended to monitor reperfusion patients. The RAMP CK-MB Assay is intended to be used only to prioritize patient management for those suspected of AMI.

Device Story

RAMP CK-MB Assay is a quantitative immunochromatographic test for measuring CK-MB levels in EDTA whole blood. User adds diluted blood to a test cartridge containing an immunochromatographic strip. Fluorescent-dyed latex particles coated with anti-CK-MB antibodies bind to CK-MB in the sample; complexes are immobilized at a detection zone. RAMP Clinical Reader measures fluorescence at the detection and internal control zones. A ratio of these values provides a quantitative concentration. Used in clinical settings to prioritize management of patients suspected of AMI. Results assist clinicians in rapid diagnostic decision-making.

Clinical Evidence

Method comparison study with 365 subjects (180 normal, 185 suspected AMI). Compared RAMP CK-MB (EDTA whole blood) against Dade Behring Dimension CK-MB (heparinized plasma). Results: Sensitivity 96.04% (95% CI: 92.24-99.84), Specificity 97.73% (95% CI: 95.93-99.53), Concordance 97.26%. Correlation R=0.986. Analytical performance included precision (total CV 4.8-8.6%) and linearity (R=0.999).

Technological Characteristics

Quantitative immunochromatographic test strip; fluorescent-dyed latex particles; anti-CK-MB antibodies. Uses RAMP Clinical Reader (fluorometer) for signal detection. Sample type: EDTA whole blood. Connectivity: Standalone reader. No specific material standards or software architecture details provided.

Indications for Use

Indicated for patients suspected of acute myocardial infarction (AMI) to aid in rapid diagnosis and prioritize patient management. Not indicated for monitoring reperfusion patients.

Regulatory Classification

Identification

A creatine phosphokinase/creatine kinase or isoenzymes test system is a device intended to measure the activity of the enzyme creatine phosphokinase or its isoenzymes (a group of enzymes with similar biological activity) in plasma and serum. Measurements of creatine phosphokinase and its isoenzymes are used in the diagnosis and treatment of myocardial infarction and muscle diseases such as progressive, Duchenne-type muscular dystrophy.

Predicate Devices

• Triage Cardiac Panel®; CK-MB Assay (K973126)
• Dimension® RxL Mass Creatine Kinase MB Isoenzyme Flex® (K970343)

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY DEVICE ONLY TEMPLATE

A. 510(k) Number:
K033747

B. Purpose for Submission:
To obtain original clearance using traditional 510(k) for this new assay.

C. Analyte:
Creatine Kinase MB isoenzyme (CK-MB)

D. Type of Test:
Immunochromatographic fluorescence immunoassay

E. Applicant:
RESPONSE BIOMEDICAL CORP.

F. Proprietary and Established Names:
RAMP CK-MB ASSAY

G. Regulatory Information:
1. Regulation section:
21CFR §862.1215 -Creatine phosphokinase/creatine kinase or isoenzymes test system.
2. Classification:
2
3. Product Code:
JHX
4. Panel:
CH

H. Intended Use:
1. Indication(s) for use:
The RAMP CK-MB Assay is a quantitative immunochromatographic test indicated for use as an in vitro diagnostic product used with the RAMP Clinical Reader to measure CK-MB levels in EDTA whole blood. Measurement of CK-MB aids in the rapid diagnosis of acute myocardial infarction (AMI).
2. Special condition for use statement(s):
The RAMP CK-MB Assay is not intended to monitor reperfusion patients. The RAMP CK-MB Assay is intended to be used only to prioritize patient management for those suspected of AMI.
3. Special instrument Requirements:
RAMP® Clinical Reader

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# I. Device Description:

The RAMP CK-MB Assay is a quantitative immunochromatographic test for the determination of CK-MB levels in EDTA whole blood. Diluted EDTA whole blood is added to the sample well of the Test Cartridge which houses the immunochromatographic test strip. The red blood cells are retained in the sample pad, and the separated plasma migrates along the strip. Fluorescent-dyed latex particles coated with anti-CK-MB antibodies bind to CK-MB, if present in the sample. As the sample migrates along the strip, CK-MB bound particles are immobilized at the detection zone, and additional particles are immobilized at the internal control zone.

The RAMP Clinical Reader then measures the amount of fluorescence emitted by the complexes bound at the detection zone and at the internal control zone. Using a ratio between the two fluorescence values, a quantitative reading is calculated.

# J. Substantial Equivalence Information:

1. Predicate device name(s):

Immunoassay: Triage Cardiac Panel®; CK-MB Assay (K973126) which is currently being marketed by Biosite Diagnostics, Inc.

Immunoassay: Dimension® RxL Mass Creatine Kinase MB Isoenzyme Flex®, (K970343) which is currently being marketed by Dade Behring Inc.

2. Predicate K number(s):

K973126

K970343

3. Comparison with predicate:

The RAMP CK-MB Assay, Triage Cardiac Panel (Triage) - CK-MB; and Dade Dimension RxL (Dimension) Creatine Kinase MB Isoenzyme (CK-MB) Flex Assays are for the quantitative measurement of CK-MB in human whole blood (RAMP and Triage) or plasma (Triage and Dimension). All three immunoassays utilize the binding of CK-MB to specific antibodies and utilize light in their respective detection systems. Both the RAMP and Triage assays measure light production from a fluorescence reaction using a fluorometer while the Dimension measures the amount of colored product produced which is directly proportional to the concentration of CK-MB present in the patient sample. Both the RAMP CK-MB and the Triage CK-MB are quantitative immunochromatographic tests, whereas the Dimension CK-MB test is a sandwich enzyme immunoassay.

An additional minor difference is that while both the RAMP CK-MB and the Dimension CK-MB assay are single tests for CK-MB determination, the Triage Cardiac Panel consists of three tests, CK-MB, Troponin I, and Myoglobin.

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| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Intended Use | The RAMP CK-MB Assay is a quantitative immunochromatographic test indicated for use as an in vitro diagnostic product used with the RAMP Clinical Reader to measure Creatine Kinase MB (CK-MB) levels in EDTA whole blood. Measurement of CK-MB aids in the rapid diagnosis of acute myocardial infarction (AMI). | Triage Cardiac Panel is a fluorescence immunoassay used for the quantitative determination of TnI, Creatine Kinase MB and Myoglobin in heparinized whole blood and plasma specimens. The test is used as an aid in the diagnosis of myocardial infarction. |
| Test Principle | Immunochromatographic fluorescence immunoassay | Same |
| Target Population | Suspected Acute Myocardial Infarction | Same |
| Test Procedure | Add sample to Test Cartridge | Same |
| Automated Processing | Instrument transport of Test Cartridge within reader only moving stepNo internal liquid handlingOne step immunochromatography assay requiring no additional washes | Same |
| Read Results | Read results on screen | Same |
| Test Time | 12 minutes after Test Components come to room temperature | 15 – 18 minutes after Test Device comes to room temperature |
| Waste Handling | Dispose of Test Cartridge as per correct institutional biohazard procedure | same |
| Automated Analysis | Yes | Yes |
| Self Contained | Yes | Yes |
| Portable | Yes | Yes |
| Battery Operation Available | Yes, rechargeable | Yes, replaceable |
| Special Instrumentation Required | Yes; RAMP Clinical Reader | Yes; Triage Meter |

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| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Sample Preparation | Use provided pipette to dilute sample 1/8 in provided pre-measured diluent vial | Use special provided transfer pipette to withdraw sample to the level of the lower bulb |
| Specimen Type | Whole anticoagulated blood (EDTA) | Whole anticoagulated blood or plasma (heparin) |
| Specimen Storage:
Ambient
2-8 °C
-20 °C | up to 2 hours
up to 2 days
not recommended | not recommended
up to 24 hours
for longer term storage (plasma only) |

K. Standard/Guidance Document Referenced (if applicable):
None Referenced

L. Test Principle:
Immunochromatographic fluorescence immunoassay

M. Performance Characteristics (if/when applicable):
1. Analytical performance:
a. Precision/Reproducibility:
The intra-assay and the inter-assay precision of the RAMP CK-MB Assay were determined by one operator assaying duplicates of three standards (7.19, 14.29 and 25.06 ng/mL CK-MB) twice each day over 10 days. The mean, standard deviation and % CV were calculated for the predicted CK-MB at each concentration.

| Precision | Standards
CK-MB Mean Concentration (ng/mL) | | |
| --- | --- | --- | --- |
| | 7.19 | 14.29 | 25.06 |
| Within Run Precision | 7.7% | 7.8% | 4.8% |
| Total Precision | 8.6% | 8.5% | 6.9% |

b. Linearity/assay reportable range:
CK-MB antigen concentrations of 2.5, 5.0, 10.0, 20.0, 40.0 and 60.0 ng/mL were prepared in normal donor EDTA blood. The linearity and percent recovery were determined by assaying five replicates of each concentration and baseline. The mean, standard deviation and %CV were calculated for the predicted CK-MB at each concentration. Linear regression analysis of actual CK-MB concentration versus expected CK-MB concentration resulted with an R = 0.999 and a slope of 1.05 with an offset of 0.098. The

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recovery of spiked CK-MB antigen at the six concentrations ranged from 99 to 111% with an average of 106%.

c. Traceability (controls, calibrators, or method):

Each RAMP CK-MB kit includes a Lot Card that is individually packaged in an anti-static pouch. The Lot Card provides information specific to the kit Test Cartridge lot, including lot number, expiration date, and standard curve information. Insertion of the Lot Card into the Reader is the only calibration necessary for each lot of reagents.

Quality Controls are provided in every Test Cartridge, which serve as built-in performance controls for routine QC requirements. A comparison of this control and the assay result indicate that sufficient sample was applied to the test device, that unbound fluorescent label washed sufficiently from the detection zone, and the device was inserted and read properly by the instrument. This control also prevents a used cartridge from being re-run by the reader. Antibody quality, system function and assay timing are checked on each assay run. An unacceptable result from the control displays a warning message on the instrument indicating that the test should be repeated.

It is recommended that a commercial external control material be run in the RAMP CK-MB Assay weekly, or in conformance with regulatory or accreditation requirements. To run a QC sample, follow the instructions under the SAMPLE ANALYSIS PROCEDURE section. Treat the control as a whole blood sample.

d. Detection limit:

The lower limit of detection (LLD) is defined as the analyte concentration corresponding to the mean (n=20) plus 2 standard deviations of the zero. The LLD is 0.32 ng/mL CK-MB. CK-MB levels in excess of 80 ng/mL are reported as greater than (>) 80 ng/mL.

e. Analytical specificity:

Potentially cross-reactive substances were evaluated by spiking different concentrations into blood. CK-MM up to 50,000 ng/mL and CK-BB up to 1000 ng/mL appear to have no cross-reactivity with the RAMP CK-MB Assay. Human anti-mouse antibodies (HAMA), human anti-goat antibodies (HAGA), human anti-rabbit antibodies (HARA) and Rheumatoid Factor (RhF) appear to have limited cross-reactivity with the RAMP CK-MB Assay.

Potentially interfering substances were evaluated by spiking different concentrations of potential interferents in blood with CK-MB added. Different blood samples were used for each potential interferent. Interference was evaluated by calculating the CK-MB concentration

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of potential interferent-spiked blood, expressed as a percentage of the CK-MB concentration of the unspiked (no potential interferent) blood sample. No evidence of cross-reactivity or interference was observed for hemoglobin, triglyceride, bilirubin, cholesterol or heparin at levels of very high physiological concentrations, up to 2000 mg/dL, 3000 mg/dL, 80 mg/dL, 500 mg/dL, and 104 IU/mL, respectively. No trend was observed in the CK-MB predictions as the concentration of potential interferent was increased.

f. Assay cut-off:
0.32 ng/mL (LLD)

2. Comparison studies:

a. Method comparison with predicate device:

Three hundred and sixty-five (365) subjects were enrolled in the Method Comparison Study. Of these subjects, 180 were normal individuals (84 males and 96 females) and 185 were suspected of acute myocardial infarct (AMI) based on the individual hospital criteria (115 males and 70 females). EDTA and heparin whole blood samples were obtained for each of these subjects. All normal subjects were consented. Waste samples were used for the subjects suspected of AMI. An aliquot of the whole blood was taken for the RAMP CK-MB Assay and heparinized plasma was prepared for the Dade Behring Dimension CK-MB Assay. The data was winsorized to account for the differing reportable ranges and two outliers were removed from the suspect AMI samples. The correlation data is presented in the table below.

| Population | n | Sy.x | Slope y = | Intercept | R |
| --- | --- | --- | --- | --- | --- |
| Combined Populations | 363 | 3.11 | 0.966 | 0.600 | 0.986 |
| Suspect AMI Subjects | 183 | 4.28 | 0.955 | 1.207 | 0.984 |

The sensitivity, specificity, and percent agreement of all samples were calculated comparing a clinical cutoff of 6.0 ng/mL CK-MB for the RAMP CK-MB Assay to the published clinical cutoff of 5.0 ng/mL CK-MB presented in the Dade Dimension package insert. The data is presented in the table below.

| | n | (%) | s.e. | 95 % CI | |
| --- | --- | --- | --- | --- | --- |
| Sensitivity | 101 | 96.04 | 1.94 | 92.24 | 99.84 |
| Specificity | 264 | 97.73 | 0.92 | 95.93 | 99.53 |
| PV+ | 103 | 94.18 | 2.31 | 89.65 | 98.70 |
| PV- | 262 | 98.47 | 0.76 | 96.99 | 99.96 |
| Concordance | 365 | 97.26 | 0.85 | 95.59 | 98.94 |

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b. Matrix comparison: Not Applicable

3. Clinical studies:

a. Clinical sensitivity: Not Applicable
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: One hundred and eighty (180) normal individuals were enrolled in the Expected Values Study. RAMP CK-MB expected normal values ranged from 0.00 to $3.74\mathrm{ng / mL}$ CK-MB. The percentile ranking is presented in the table below.

| Percentile | ng/mL |
| --- | --- |
| 5th(LLN) | 0.00 |
| 50th | 0.78 |
| 90th | 2.87 |
| 95th(ULN) | 3.74 |
| 97.5th | 4.99 |

The RAMP CK-MB Assay is not intended to monitor reperfusion patients. The RAMP CK-MB Assay is intended to be used only to prioritize patient management for those suspected of AMI.

# N. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.