The CKMB Test is an in vitro diagnostic assay for the quantitative determination of creatine kinase isoform MB in EDTA or lithium heparin whole-blood or plasma specimens on the AQT90 FLEX analyzer in point of care and laboratory settings. It is intended for use as an aid in the diagnosis of myocardial infarction.

The Myo Test is an in vitro diagnostic assay for the quantitative determination of myoglobin in EDTA or lithium heparin whole-blood or plasma specimens on the AQT90 FLEX analyzer in point of care and laboratory settings. It is intended for use as an aid in the rapid diagnosis of heart disease, for example, acute myocardial infarction.

For in vitro diagnostic use. The AQT90 FLEX analyzer is an immunoassay instrument based on the quantitative determination of time-resolved fluorescence to estimate the concentrations of clinically relevant markers on whole-blood and plasma specimens to which a relevant anticoagulant has been added. It is intended for use in point-of-care and laboratory settings.

Device Description

The AQT90 FLEX is a cartridge-based immunoassay analyzer, based on time-resolved fluorescence using a europium (Eu) chelate as the fluorescent label. The test receptacles for the assay are test cups, which contain the antibodies used for capture of the analyte, and the Eu chelate labeled antibodies used to trace the captured analyte. The sample is added to the test cup together with assay buffer. The cup is then incubated to allow formation of the immuno-complex, and subsequently washed to remove unbound antibodies and sample material. Finally, the cup is exposed to excitation light, and after a delay the emitted light generated by the fluorescent label is measured by single photon counting. The total count is then compared to an assay calibration curve to obtain a quantitative measurement of the analyte's concentration in the sample.

This technology uses dried reagents deposited in the test cups and in the calibration adjustment cups – no liquids other than the sample itself together with the assay buffer are required.

AI/ML Overview

The provided document is a 510(k) premarket notification from Radiometer Medical ApS for their AQT90 FLEX CKMB Test Kit, AQT90 FLEX Myo Test Kit, and the AQT90 FLEX analyzer. The submission is to seek clearance for modifications to the existing AQT90 FLEX system devices.

The document does not describe a study involving an AI model or a human-in-the-loop performance study. Instead, it describes analytical performance studies of in-vitro diagnostic assays (Myoglobin and CKMB) on a laboratory analyzer. Therefore, many of the requested elements pertaining to AI models, human experts, ground truth adjudication, MRMC studies, and training datasets are not applicable to this document.

However, I can extract information related to the acceptance criteria (implicitly, the performance metrics evaluated) and the studies conducted to prove the device meets these criteria in the context of an in-vitro diagnostic device.

Here's a breakdown of the available information:

1. Table of Acceptance Criteria (Performance Metrics) and Reported Device Performance

For an in-vitro diagnostic device, acceptance criteria are typically related to analytical performance characteristics such as linearity, limits of detection/quantitation, method comparison (agreement with a predicate), and precision. The reported device performance is the outcome of the studies conducted for these characteristics.

AQT90 FLEX Myo Test Kit

Performance Metric (Implicit Acceptance Criteria)	Reported Device Performance
Linearity (within 10% non-linearity)	Whole Blood & Plasma: Linear within the reportable range (20 – 900 ng/mL), with non-linearity within 10%.
Limit of Blank (LoB)	Myo: 0.5 ng/mL (µg/L)
Limit of Detection (LoD)	Myo: 1 ng/mL (µg/L)
Limit of Quantitation (LoQ) (at 10% CVWithin-lab)	Myo: 1 ng/mL (µg/L)
Method Comparison (vs. predicate) (Implicitly, good correlation and minimal bias)	Plasma: y = 1.01 x - 0.14 (n=103; r²=1.0) compared to predicate.
Matrix Comparison (Implicitly, interchangeability of matrix types)	Equivalence confirmed: No significant matrix effect differences between whole blood vs. plasma or lithium heparin vs. EDTA. (Regression equations provided for various comparisons, all showing strong correlation (r²=1.0) and slopes close to 1).
Precision (Repeatability, Between-Run, Total CVs)	Myo Whole Blood: L1 (57 ng/mL): Total CV 2.3% L2 (92 ng/mL): Total CV 3.0% L3 (622 ng/mL): Total CV 2.4%Myo Plasma: L1 (53 ng/mL): Total CV 2.1% L2 (95 ng/mL): Total CV 2.2% L3 (586 ng/mL): Total CV 2.2%

AQT90 FLEX CKMB Test Kit

Performance Metric (Implicit Acceptance Criteria)	Reported Device Performance
Linearity (within 10% non-linearity)	Whole Blood & Plasma: Linear within the reportable range (1.5 – 300 ng/mL), with non-linearity within 10%.
Limit of Blank (LoB)	CKMB: 0.5 ng/mL (µg/L)
Limit of Detection (LoD)	CKMB: 1 ng/mL (µg/L)
Limit of Quantitation (LoQ) (at 20% CVWithin-lab)	CKMB: 1 ng/mL (µg/L)
Method Comparison (vs. predicate) (Implicitly, good correlation and minimal bias)	Plasma: y = 0.99 x - 0.18 (n=107; r²=1.0) compared to predicate.
Matrix Comparison (Implicitly, interchangeability of matrix types)	Equivalence confirmed: No significant matrix effect differences between whole blood vs. plasma or lithium heparin vs. EDTA. (Regression equations provided for various comparisons, all showing strong correlation (r²=1.0) and slopes close to 1).
Precision (Repeatability, Between-Run, Total CVs)	CKMB Whole Blood: L1 (2.6 ng/mL): Total CV 4.8% L2 (14 ng/mL): Total CV 4.9% L3 (204 ng/mL): Total CV 3.4%CKMB Plasma: L1 (2.3 ng/mL): Total CV 3.7% L2 (8.4 ng/mL): Total CV 2.8% L3 (209 ng/mL): Total CV 2.2%

2. Sample Size Used for the Test Set and Data Provenance

Linearity (Myo & CKMB): 11 sample levels for linearity series, measured with 10 replicates each. This is an in vitro analytical study, not patient data.
LoB/LoD/LoQ (Myo & CKMB):
- LoB: Four blank samples measured with 5 replicates on 3 days, using 2 test kit lots and 2 analyzers. Total 60 measurements per test kit lot.
- LoD/LoQ: 10 samples per matrix (lithium heparin whole blood and plasma).
Method Comparison (Myo & CKMB):
- Myo: n=103 lithium heparin plasma samples.
- CKMB: n=107 lithium heparin plasma samples.
Matrix Comparison (Myo & CKMB):
- Myo: n=125 for most comparisons (e.g., Liph/Pl vs Liph/WB), some n=127. Paired lithium heparin and EDTA specimens.
- CKMB: n=106 for Liph/Pl vs Liph/WB, others n=104, 103, 101. Paired lithium heparin and EDTA specimens.
Precision (Myo & CKMB):
- Whole Blood: 3 lithium heparin whole blood samples, measured 5 times five replicates (total 25 measurements per sample level).
- Plasma: 3 lithium heparin plasma pools, measured across 20 test days, twice a day with 2 replicates (total 80 measurements per sample level).

Data Provenance: The studies were conducted "at one internal test site" for method comparison and "at three hospital laboratory sites" for matrix comparison. This indicates domestic (likely Denmark, where the manufacturer is located) or potentially international clinical laboratory settings. The data are prospective in the sense that they were generated specifically for these validation studies using prepared samples (diluted native specimens, spiked specimens, blank samples). They are not patient-outcome data or retrospective chart reviews.

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications of those Experts:

Not Applicable. For an in-vitro diagnostic device measuring analytes (myoglobin, CK-MB), the "ground truth" isn't established by human experts in the same way as, for example, image interpretation. The ground truth for these studies is the reference measurement from the original (predicate) device or the known concentration of prepared analytical samples.

4. Adjudication Method for the Test Set:

Not Applicable. No human interpretation or adjudication is involved in determining the concentration of analytes in a blood sample by an immunoassay method.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:

No. This type of study is relevant for AI systems that assist human readers in tasks like image interpretation. This document describes the analytical performance of an in-vitro diagnostic device that quantitatively measures biochemical markers. There are no human readers or AI assistance involved in this context.

6. Standalone (Algorithm Only Without Human-in-the-Loop Performance):

Yes, in the context of an IVD. The performance data presented (linearity, LoD/LoQ, method comparison, precision) are all "standalone" in the sense that they demonstrate the analytical performance of the AQT90 FLEX system (analyzer and test kits) independent of human interpretation or intervention beyond proper sample handling and instrument operation. This isn't an "algorithm only" in the AI sense, but rather the performance of analytical machines.

7. The Type of Ground Truth Used:

Reference Measurement/Known Concentration:
- For Linearity, LoB/LoD/LoQ, and Precision: The ground truth is established by preparing samples with known or precisely characterized concentrations of the analytes (e.g., diluted native specimens, spiked specimens, blank samples).
- For Method Comparison: The ground truth is the measurement obtained from the predicate device (the previously cleared version of the AQT90 FLEX system devices). The goal is to show agreement between the modified device and the predicate.

8. The Sample Size for the Training Set:

Not Applicable. This document describes the validation of an in-vitro diagnostic device, not an AI model. Therefore, there's no "training set" in the machine learning sense. The device performs a chemical reaction and optical measurement based on established immunoassay principles, not a learned algorithm trained on data.

9. How the Ground Truth for the Training Set was Established:

Not Applicable. As there is no training set for an AI model, this question is not relevant.

Summary

{0}------------------------------------------------

Image /page/0/Picture/1 description: The image shows the seal of the U.S. Department of Health & Human Services. The seal features a stylized caduceus, a symbol often associated with medicine and healthcare, with three intertwined snakes and wings. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the caduceus.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002 September 21, 2017

RADIOMETER MEDICAL APS PER PAPE THOMSEN REGULATORY AFFAIRS SPECIALIST AKANDEVEJ 21 2700 BROENSHOEJ, DENMARK

Re: K163462 Trade/Device Name: AQT90 FLEX CKMB Test Kit AQT90 FLEX Myo Test Kit AQT90 FLEX Regulation Number: 21 CFR 862.1215 Regulation Name: Creatine phosphokinase/creatine kinase or isoenzymes test system Regulatory Class: II Product Code: JHX, DDR, KHO Dated: September 15, 2017 Received: September 18, 2017

Dear Per Pape Thomsen:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA), You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

{1}------------------------------------------------

electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Kellie B. Kelm -S

for Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

{2}------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

510(k) Number (if known) K163462

Device Name AQT90 FLEX CKMB Test Kit

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the
time to review instructions, search existing data sources, gather and maintain the data needed and complete
and review the collection of information. Send comments regarding this burden estimate or any other aspect
of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services

Food and Drug Administration

Office of Chief Information Officer

Paperwork Reduction Act (PRA) Staff

PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

Form Approved: OMB No. 0910-0120 Expiration Date: 06/30/2020 See PRA Statement below.

{3}------------------------------------------------

Indications for Use

510(k) Number (if known) K163462

Device Name AQT90 FLEX Myo Test Kit

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
☒ Prescription Use (Part 21 CFR 801 Subpart D) ☐ Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

Department of Health and Human Services

Food and Drug Administration

Office of Chief Information Officer

Paperwork Reduction Act (PRA) Staff

PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

Form Approved: OMB No. 0910-0120 Expiration Date: 06/30/2020 See PRA Statement below.

{4}------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

510(k) Number (if known) K163462

Device Name AQT90 FLEX

Indications for Use (Describe)

For in vitro diagnostic use.

The AQT90 FLEX analyzer is an immunoassay instrument based on the quantitative determination of time-resolved fluorescence to estimate the concentrations of clinically relevant markers on whole-blood and plasma specimens to which a relevant anticoagulant has been added. It is intended for use in point-of-care and laboratory settings.

Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)
----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------	-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

Department of Health and Human Services
Food and Drug Administration

Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

Type of Use (Select one or both, as applicable)

{5}------------------------------------------------

K163462

510(k) Summary

This summary of 510(k) safety and effectiveness information is submitted in accordance with the requirements of 21 CFR 807.92.

1. Submitter and Contact Information

Submitter
Company Name:	Radiometer Medical ApS
ER Number:	3002807968
Address:	Aakandevej 212700 BroenshoejDenmark
Phone:	+45 3827 3827
Fax:	+45 3827 2727
Contact Person
Name:	Per Pape Thomsen
Function:	Regulatory Affairs Specialist
E-mail:	per.pape.thomsen@radiometer.dk
Phone:	+45 3827 3304
Fax:	+45 3827 2727

Date prepared: September 18, 2017

2.a Device Information

Device Name:	AQT90 FLEX Myo Test Kit
Regulation Name:	Myoglobin Immulogical Test System
Product Code:	DDR
Regulation Section:	21 CFR 866.5680
Classification:	Class II
Classification Panel:	Immunology (82)
Device Name:	AQT90 FLEX CKMB Test Kit
Regulation Name:Product Code:	Creatine phosphokinase/creatine kinase or isoenzymes test systemJHX
Regulation Section:	21 CFR 862.1215
Classification:	Class II
Classification Panel:	Clinical Chemistry (75)
Device Name:	AQT90 FLEX
Regulation Name:	Fluorometer for clinical use
Product Code:	KHO
Regulation Section:	21 CFR 862.2560
Classification:	Class I
Classification Panel:	Clinical Chemistry (75)

{6}------------------------------------------------

2.b Device Description

This technology uses dried reagents deposited in the test cups and in the calibration adjustment cups – no liquids other than the sample itself together with the assay buffer are required. In summary, the procedure is as follows:

1. Metering of an exact amount of sample and assay buffer and dispensing into a test cup
1. Incubating test cup
1. Washing of the test cup to remove unbound tracer antibodies and sample material
1. Drying the test cup
1. Measuring

2.c Purpose of submission

The purpose of this submission is to seek clearance for modifications to the existing AQT90 FLEX system devices.

The modifications implemented on the AQT90 FLEX System are outlined below:

Three separate analyzer manuals are combined into one manual, which is updated to ● include changes in analyzer functionality
. System Clean is updated to implement semi-automated System Clean using two new accessories; Blank Cartridge and Cleaning Solution Tubes
. Algorithm used to determine hematocrit value is updated
. Needle used in cup wash is shortened
Flash lamp in detection system is changed to a new flash lamp
The filter, which is located in the liquid pathway after the cup washing step, is removed ●
Test cartridge and CAL cartridge foil is pre-laser cut on one side of the foil instead of both . sides
. Recombinant streptavidin from a new supplier which is chemical to the old native streptavidin. Streptavidin is used to bind capture antibody onto the cup surface

These modifications include changes which potentially could impact Myo Test and CKMB Test performance and instructions for use.

In addition to the modifications listed above the AQT90 FLEX analyzer has been maintained via software updates. Also minor updates to consumable inserts have been introduced.

{7}------------------------------------------------

3. Indications for Use

AQT90 FLEX

For in vitro diagnostic use.

The AQT90 FLEX analyzer is an immunoassay instrument based on the quantitative determination of time-resolved fluorescence to estimate the concentrations of clinically relevant markers on whole-blood and plasma specimens to which a relevant anticoaqulant has been added. It is intended for use in point-of-care and laboratory settings.

AQT90 FLEX Myo Test Kit

AQT90 FLEX CKMB Test Kit

4. Substantial Equivalence

The AQT90 FLEX System devices are substantial equivalent in intended use, fundamental scientific technology, and characteristics to the predicates (K112161, K120326).

Predicate devices

Predicate name	Device Manufacturer	510(k) number
AQT90 FLEX Myo Test kit	Radiometer Medical Aps	K112161
AQT90 FLEX CKMB Test Kit	Radiometer Medical Aps	K120326
AQT90 FLEX (analyzer)	Radiometer Medical Aps	K112161

Similarities and Differences

The updated AQT90 FLEX System devices are compared to the predicates in the tables below.

Characteristic	Predicate device (K112161)AQT90 FLEX Myo Test kit	AQT90 FLEX Myo Test kit(updated)
Intended use	AQT90 FLEX Myo Test is an invitro diagnostic assay for thequantitative determination ofmyoglobin in EDTA or lithium-heparin whole blood or plasmaspecimens on the AQT90 FLEXanalyzer in point of care andlaboratory settings. It is indicated foruse as an aid in the rapid diagnosisof heart disease, e.g. acutemyocardial infarction.	Same

{8}------------------------------------------------

Characteristic	Predicate device (K112161)AQT90 FLEX Myo Test kit	AQT90 FLEX Myo Test kit(updated)
Principle	Quantitative time-resolvedfluorimetric one-step sandwichimmunoassay.	Same
Traceability	Scripps M0725	Same
Reportable rangeng/mL (µg/L)	20 to 900	Same
Cartridge foil	Test cartridge is pre-laser cut onboth sides of the foil.	Test cartridge is pre-laser cut onone side of the foil.

Characteristic	Predicate device (K120326)AQT90 FLEX CKMB Test Kit	AQT90 FLEX CKMB Test Kit(updated)
Intended use	AQT90 FLEX CKMB Test is an in vitro diagnostic assay for thequantitative determination ofcreatine kinase isoform MB in EDTAor lithium-heparin whole blood orplasma specimens on the AQT90FLEX analyzer in point of care andlaboratory settings. It is intended foruse as an aid in the diagnosis ofmyocardial infarction.	Same
Principle	Quantitative time-resolvedfluorimetric one-step sandwichimmunoassay	Same
Traceability	ERM-AD455/IFCC	Same
Reportable rangeng/mL (µg/L)	1.5 to 300	Same
Cartridge foil	Test cartridge is pre-laser cut onboth sides of the foil.	Test cartridge is pre-laser cut onone side of the foil.

{9}------------------------------------------------

Characteristic	Predicate device (K112161)AQT90 FLEX (analyzer)	AQT90 FLEX (analyzer)(updated)
Intended use	AQT90 FLEX analyzer is for in vitrodiagnostic use. The AQT90 FLEXanalyzer is an immunoassayinstrument based on the quantitativedetermination of time-resolvedfluorescence to estimate theconcentrations of clinically relevantmarkers on whole-blood and plasmaspecimens to which a relevantanticoagulant has been added. It isintended for use in point of care andlaboratory settings.	Same
System cleaning	System cleaning using customerprepared cleaning sample and testcup.	Semi-automated system cleaningusing cleaning tube from CleaningSolution Tubes and blank cup fromBlank Cartridge.

5. Performance Characteristics

No performance data are affected by the introduction of the modifications to the AQT90 FLEX system. The existing performance data still apply.

To confirm that the modifications have no effect on performance the following studies for Myo test and CKMB test were conducted: Linearity, LoB/LoQ/LoQ, Method Comparison, Matrix comparison, Precision.

5.1 Myo Test

Myo Linearity

The study was designed according to CLSI quideline EP06-A. Low and High concentration lithium heparin whole blood and lithium heparin plasma samples were used to prepare a linearity series of 11 sample levels. The samples were measured with ten replicates in random order using one Test Kit lot and one analyzer during one day. The linear, quadratic and cubic regression models were fit to the data weighted by 1/variance. The statistical significance of each fit parameter was determined and, where appropriate, the degree of non-linearity was calculated.

The degree of non-linearity was determined to be within 10 % throughout the reportable range (20 – 900 ng/mL) for whole blood and plasma.

{10}------------------------------------------------

Myo LoB/LoD/LoQ

The study was designed according to CLSI guideline EP17-A2.

The determination of LoB was performed by measuring four blank samples with five replicates on three days using two Test Kit lots and two AQT90 FLEX analyzers. The total number of measurements per Test Kit lot across all samples and days was 60.

The determination of LoD and LoQ was performed with lithium heparin whole blood and lithium heparin plasma. Ten samples per matrix were measured using two Test Kit lots and two AQT90 FLEX analyzers. The LoB estimate, determined by nonparametric method, and the SDwittin-lab of the lowest concentration samples were used to determine the LoD. A power function was fit to the concentration versus CV withing data from all samples. The intersection of this fit and 10 % CVwithin-lab was used to determine LoQ.

The results of the studies support the claimed LoB, LoQ summarized in the table below

Analytical sensitivity	Myo ng/mL (µg/L)
Limit of blank (LoB)	0.5
Limit of detection (LoD)	1
Limit of quantitation (LoQ)	1

Myo Method Comparison

The study was designed according to CLSI quideline EPO9-A3. Lithium heparin plasma samples in the range 26 - 897 ng/mL were measured at one internal test site across four AQT90 FLEX analyzers. One Myo Test Kit lot was used. The Myoglobin measured on the modified AQT90 FLEX analyzer (y) was compared to the Myoglobin measured on the predicate (x).

The Passing-Bablok reqression equation for plasma was found to be: y= 1.01 x - 0.14 (n= 103; r2= 1.0)

Myo Matrix Comparison

The study was designed according to CLSI quideline EP09-A3. The study was conducted at three hospital laboratory sites on one AQT90 FLEX analyzer per site. One Myo Test Kit lot was used across sites. Paired lithium heparin and EDTA specimens were measured across the reportable range of 20 - 900 ng/mL.

The equivalence between the measurement procedures on the AQT90 FLEX analyzer with lithium heparin whole blood, lithium heparin plasma, EDTA whole blood, and EDTA plasma samples were determined. No matrix effect differences were seen caused by differences in specimen types (whole blood vs. plasma) or anticoaqulants (lithium heparin vs. EDTA). The four anticoagulant matrix combinations can be used interchangeably.

The Passing-Bablok regression equations were found to be:

Lithium heparin plasma (y) = 0.99 Lithium heparin whole blood (x) - 1.0 (n= 125; r²= 1.0) EDTA plasma (y) = 0.96 EDTA whole blood (x) - 1.4 (n= 125; r2= 1.0) EDTA whole blood (y) = 1.01 Lithium heparin whole blood (x) + 0.8 (n= 127; r == 1.0) EDTA plasma (y) = 0.99 Lithium heparin plasma (x) + 0.2 (n= 124; r²= 1.0) EDTA whole blood (y) = 1.03 Lithium heparin plasma (x) + 1.6 (n= 125; r²= 1.0)

{11}------------------------------------------------

Myo Precision

The study was designed according to CLSI guideline EP05-A3.

In the whole blood precision study three lithium heparin whole blood samples were measured, each within one day five times (runs) with five replicates using one analyzer and one Test Kit lot.

Myo whole blood precision results; L1: diluted native specimen, L2: native specimen, L3: native specimen spiked with antigen.

			Repeatability		Between-Run		Total
Sampledescription	Mean value(ng/mL)	n	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)
L1	57	25	1.30	2.3	0.00	0.0	1.30	2.3
L2	92	25	2.57	2.8	0.93	1.0	2.73	3.0
L3	622	25	14.1	2.3	4.94	0.8	15.0	2.4

In the plasma precision study three lithium heparin plasma pools were measured across 20 test days, twice a day with two replicates, using one analyzer and one Test Kit lot.

Myo plasma precision results; L1: native specimen pool, L2: native specimen pool, L3: native specimen pool spiked with antigen.

Sampledescription	Meanvalue(ng/mL)	n	Repeatability		Between-Run		Between-Day		Total
			SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)
L1	53	80	1.0	2.0	0.47	0.9	0.00	0.0	1.1	2.1
L2	95	80	1.9	2.0	0.00	0.0	0.84	0.9	2.1	2.2
L3	586	80	13	2.2	0.00	0.0	0.00	0.0	13	2.2

5.2 CKMB Test

CKMB Linearity

The study was designed according to CLSI guideline EP06-A. Low and High concentration lithium heparin whole blood and lithium heparin plasma samples were used to prepare a linearity series of 11 sample levels. The samples were measured with ten replicates in random order using one Test Kit lot and one analyzer during one day. The linear, quadratic and cubic regression models were fit to the data weighted by 1/variance. The statistical significance of each fit parameter was determined and, where appropriate, the degree of non-linearity was calculated.

The degree of non-linearity was determined to be within 10 % throughout the reportable range (1.5 – 300 ng/mL) for whole blood and plasma.

CKMB LoB/LoD/LoQ

The study was designed according to CLSI guideline EP17-A2.

{12}------------------------------------------------

The determination of LoD and LoQ was performed with lithium heparin whole blood and lithium heparin plasma. Ten samples per matrix were measured using two Test Kit lots and two AQT90 FLEX analyzers. The LoB estimate, determined by nonparametric method, and the SDwittin-lab of the lowest concentration samples were used to determine the LoD. A power function was fit to the concentration versus CV within-lab plot using data from all samples. The intersection of this fit and 20 % CVwithin-lab was used to determine LoQ.

The results of the studies support the claimed LoB, LoQ summarized in the table below

Analytical sensitivity	CKMB ng/mL (µg/L)
Limit of blank (LoB)	0.5
Limit of detection (LoD)	1
Limit of quantitation (LoQ)	1

CKMB Method Comparison

The study was designed according to CLSI guideline EP09-A3. Lithium heparin plasma samples in the range 1.5 - 296 ng/mL were measured at one internal test site across four AQT90 FLEX analyzers. One CKMB Test Kit lot was used. The CKMB measured on the modified AQT90 FLEX analyzer (y) was compared to the CKMB measured on the predicate (x).

The Passing-Bablok regression equation for plasma was found to be: y= 0.99 x - 0.18 (n= 107; r2= 1.0)

CKMB Matrix Comparison

The study was designed according to CLSI guideline EP09-A3. The study was conducted at three hospital laboratory sites on one AQT90 FLEX analyzer per site. One CKMB Test Kit lot was used across sites. Paired lithium heparin and EDTA specimens were measured across the reportable range of 1.5 - 300 nq/mL.

The equivalence between the measurement procedures on the AQT90 FLEX analyzer with lithium heparin whole blood, lithium heparin plasma, EDTA whole blood, and EDTA plasma samples were determined. No matrix effect differences were seen caused by differences in specimen types (whole blood vs. plasma) or anticoagulants (lithium heparin vs. EDTA). The four anticoagulant matrix combinations can be used interchangeably.

The Passing-Bablok regression equations were found to be:

Lithium heparin plasma (y) = 0.99 Lithium heparin whole blood (x) + 0.01 (n= 106; r²= 1.0) EDTA plasma (y) = 0.99 EDTA whole blood (x) - 0.02 (n= 104; r²= 1.0) EDTA whole blood (y) = 1.02 Lithium heparin whole blood (x) + 0.01 (n= 103; r²= 1.0) EDTA plasma (y) = 1.01 Lithium heparin plasma (x) - 0.04 (n= 101; r² = 1.0) EDTA whole blood (y) = 1.03 Lithium heparin plasma (x) - 0.02 (n= 104; r = 1.0)

CKMB Precision

The study was designed according to CLSI guideline EP05-A3.

In the whole blood precision study three lithium heparin whole blood samples were measured, each within one day five times (runs) with five replicates using one analyzer and one Test Kit lot.

{13}------------------------------------------------

	Mean		Repeatability		Between-Run		Total
Sampledescription	value(ng/mL)	n	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)
L1	2.6	25	0.13	4.8	0.00	0.0	0.13	4.8
L2	14	25	0.50	3.5	0.49	3.4	0.70	4.9
L3	204	25	6.9	3.4	0.00	0.0	6.9	3.4

CKMB whole blood precision results; L1: diluted native specimen, L2: native specimen spiked with antigen, L3: native specimen spiked with antigen.

In the plasma precision study three lithium heparin plasma pools were measured across 20 test days, twice a day with two replicates, using one analyzer and one Test Kit lot.

CKMB plasma precision results; L1: native specimen pool, L2: native specimen pool spiked with high patient specimen, L3: native specimen pool spiked with antigen.

Sampledescription	Meanvalue(ng/mL)	n	Repeatability		Between-Run		Between-Day		Total
			SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)	SD(ng/mL)	CV(%)
L1	2.3	80	0.080	3.5	0.026	1.1	0.00	0.0	0.084	3.7
L2	8.4	80	0.21	2.5	0.083	1.0	0.088	1.0	0.24	2.8
L3	209	80	3.9	1.9	1.4	0.7	1.8	0.9	4.6	2.2

6. Conclusion

Based on the substantial equivalence comparison and the results of the conducted performance evaluations it has been concluded that the AQT90 FLEX System devices is as safe and effective as the predicate devices.

Regulation Number and Section

§ 862.1215 Creatine phosphokinase/creatine kinase or isoenzymes test system.

(a)
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.(b)
Classification. Class II.