K Number
K152245
Device Name
ONLINE TDM Vancomycin Gen.3
Date Cleared
2016-01-08

(151 days)

Product Code
Regulation Number
862.3950
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP Authorized
Intended Use
In vitro test for the quantitative determination of vancomycin in serum and plasma on Roche/Hitachi cobas c systems. A vancomycin test system is a device intended to measure vancomycin, an antibiotic drug, in serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy.
Device Description
The ONLINE TDM Vancomycin Gen.3 is a two reagent assay for the in vitro quantitative determination of vancomycin in human serum or plasma on automated clinical chemistry analyzers. It is a homogeneous microparticle agglutination immunoassay based on the kinetic interaction of microparticles in solution (KIMS). A competitive reaction takes place between the drug conjugate and vancomycin in the serum sample for binding to the vancomycin antibody on the microparticles. The resulting kinetic interaction of microparticles is indirectly proportional to the amount of drug present in the sample.
More Information

K060373/A001

No
The description details a homogeneous microparticle agglutination immunoassay based on kinetic interaction, which is a standard biochemical method and does not mention or imply the use of AI/ML.

No

Explanation: This device is an in vitro diagnostic test designed to measure the amount of vancomycin in a patient's serum or plasma. It provides information for diagnosis and treatment monitoring, but it does not directly apply or administer therapy to a patient.

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the measurements obtained by this device "are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy." This directly indicates a diagnostic purpose.

No

The device description clearly states it is a "two reagent assay" and a "homogeneous microparticle agglutination immunoassay," indicating it is a chemical and biological test kit, not a software-only device. It is designed to be used on automated clinical chemistry analyzers, which are hardware devices.

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

  • Intended Use: The "Intended Use / Indications for Use" section explicitly states it is an "In vitro test for the quantitative determination of vancomycin in serum and plasma". This directly aligns with the definition of an in vitro diagnostic device, which is used to examine specimens taken from the human body.
  • Device Description: The description details a "two reagent assay for the in vitro quantitative determination of vancomycin in human serum or plasma". This further confirms its use in testing biological samples outside of the body.
  • Performance Studies: The "Summary of Performance Studies" describes evaluations using "native human serum samples of patients taking Vancomycin", which are biological specimens.

The entire description points to a device designed to analyze biological samples (serum and plasma) in a laboratory setting to provide information about a patient's health status (vancomycin levels). This is the core function of an IVD.

N/A

Intended Use / Indications for Use

In vitro test for the quantitative determination of vancomycin in serum and plasma on Roche/Hitachi cobas c systems.

A vancomycin test system is a device intended to measure vancomycin, an antibiotic drug, in serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy.

Product codes

LEH

Device Description

The ONLINE TDM Vancomycin Gen.3 is a two reagent assay for the in vitro quantitative determination of vancomycin in human serum or plasma on automated clinical chemistry analyzers. It is a homogeneous microparticle agglutination immunoassay based on the kinetic interaction of microparticles in solution (KIMS). A competitive reaction takes place between the drug conjugate and vancomycin in the serum sample for binding to the vancomycin antibody on the microparticles. The resulting kinetic interaction of microparticles is indirectly proportional to the amount of drug present in the sample.

Mentions image processing

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Mentions AI, DNN, or ML

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Input Imaging Modality

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Anatomical Site

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Indicated Patient Age Range

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Intended User / Care Setting

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Description of the training set, sample size, data source, and annotation protocol

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Description of the test set, sample size, data source, and annotation protocol

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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Non-Clinical Performance Evaluation:

  • Detection Limit: LoB, LoD, and LoQ studies were performed based upon CLSI EP17-A2.
    • LoB: Concentration below which analyte-free samples are found with a probability of 95%. Diluent measured with 5-fold determinations per run on two instruments (or one for P2/P3 lots). Six runs over 3 days. Data analysis based on 95th percentile of 60 measured values.
    • LoD: Lowest analyte concentration which can be detected (value above Limit of Blank with a probability of 95%). Five serum samples with low analyte content spiked with Vancomycin measured 1-fold (or 2-fold for P2/P3 lots) per run on two instruments (or one). Six runs over 3 days.
    • LoQ: Lowest amount of analyte in a sample that can be detected quantitatively within specified precision and accuracy ranges. Fifteen serum samples prepared covering concentration range between LoB and 2x LoQ, tested in two aliquots. Six runs over 4 days. Expected value determined with Vancomycin LCMS/MS.
    • Results: LoB claimed 1.0 µg/mL, LoD claimed 1.5 µg/mL, LoQ claimed 4.0 µg/mL.
  • Precision: According to CLSI EP5-A2. Two runs per day for >= 21 days on the same analyzer. Repeatability and intermediate precision calculated.
    • Repeatability Precision Summary: CVs range from 2.3% to 8.2%.
    • Intermediate Precision Summary: CVs range from 2.9% to 10.5%.
  • Linearity: According to CLSI EP6-A. Dilution series prepared from human serum sample pool and diluent. Sixteen levels to span measuring range. Process repeated for plasma samples.
    • Results: Serum: y=1.000x-0.000, Pearson correlation coefficient (R)=0.9985 for 4.0 to 80.0 µg/mL. Plasma: y=1.000x-0.000, Pearson correlation coefficient (R)=0.9976 for 4.0 to 80.0 µg/mL.
  • Matrix Comparison - Anticoagulants: Each pair of serum and plasma of a single donor spiked with Vancomycin. 67 full tubes and 9 (or 10 for K2-EDTA) half-filled tubes included. Serum used as reference.
    • Results: Serum vs. Li-heparin: y = 1.01x -0.3, r = 0.996. Serum vs. K2-EDTA: y = 0.99x -0.0, r = 0.996. Serum vs. K3-EDTA: y = 1.00x - 0.3, r = 0.995.
  • Interferences - H, L and I Indices: Effect of endogenous interfering substances determined at two Vancomycin concentrations. Interfering substances evaluated: Hemolysis (up to H index of 1000), Lipemia (up to L index of 1000), Icterus/Bilirubin (up to I index of 60). Results compared to expected (aliquot with no interfering substance).
    • Results: No significant interference up to 60 I Index for conjugated and unconjugated bilirubin, 1000 H Index for hemolysis, and 1000 L Index (or 1000 mg/dL triglycerides).
  • Interferences - Drugs: Pharmaceutical compounds spiked into samples with high or low Vancomycin concentration according to EP7-A2 or higher.
    • Results: A table lists 17 drugs and their highest concentrations shown not to interfere, e.g., Acetylsalicylic acid 1000 mg/L, Acetaminophen 200 mg/L, Heparin 5000 U/L.
  • Method Comparison to Predicate: One hundred twenty five single native human serum samples of patients taking Vancomycin covering the reportable range were tested. Eight samples spiked, 1 sample diluted. All samples tested for icteric, lipemic and hemolytic interference. Samples tested in singlicate on candidate and predicate devices.
    • Results: Passing Bablok Regression analysis: y = 0.993x + 0.641, r = 0.994.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

  • Detection Limit:
    • LoB = 1.0 µg/mL
    • LoD = 1.5 µg/mL
    • LoQ = 4.0 µg/mL
  • Precision (CV):
    • Repeatability CV: 2.3% - 8.2%
    • Intermediate Precision CV: 2.9% - 10.5%
  • Linearity (Pearson correlation coefficient):
    • Serum: R = 0.9985
    • Plasma: R = 0.9976
  • Matrix Comparison (r):
    • Serum vs. Li-heparin: r = 0.996
    • Serum vs. K2-EDTA: r = 0.996
    • Serum vs. K3-EDTA: r = 0.995
  • Method Comparison (r):
    • r = 0.994

Predicate Device(s)

ONLINE TDM Vancomycin, K060586

Reference Device(s)

K060373/A001

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

§ 862.3950 Vancomycin test system.

(a)
Identification. A vancomycin test system is a device intended to measure vancomycin, an antibiotic drug, in serum. Measurements obtained by this device are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy.(b)
Classification. Class II.

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, stacked on top of each other. The profiles are connected by a flowing line that resembles a wave or a ribbon.

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

January 8, 2016

ROCHE DIAGNOSTICS OPERATIONS (RDO) BARBARA MCWHORTER REGULATORY AFFAIRS PRINCIPAL 9115 HAGUE ROAD INDIANAPOLIS IN 46250

Re: K152245

Trade/Device Name: ONLINE TDM Vancomvcin Gen.3 Regulation Number: 21 CFR 862.3950 Regulation Name: Vancomycin test system Regulatory Class: II Product Code: LEH Dated: December 9, 2015 Received: December 10, 2015

Dear Barbara Mc Whorter:

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

1

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.

Courtney H. Lias -S

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

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Indications for Use

510(k) Number (if known) K152245

Device Name ONLINE TDM Vancomycin Gen.3

Indications for Use (Describe)

In vitro test for the quantitative determination of vancomycin in serum and plasma on Roche/Hitachi cobas c systems.

A vancomycin test system is a device intended to measure vancomycin, an antibiotic drug, in serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy.

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)

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510(k) Summary

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

Submitter NameRoche Diagnostics Operations (RDO)
Address9115 Hague Road
Indianapolis, IN, 46250, USA
ContactBarbara McWhorter
Phone: (317) 521-2336
FAX: (317) 521-2324
Email: Barbara.McWhorter@roche.com
Date PreparedJuly 31st, 2015
Proprietary NameONLINE TDM Vancomycin Gen.3
Common NameImmunoassay, Vancomycin
Classification NameClinical Toxicology Test Systems, Class II
Product CodesLEH, 21 CFR § 862.3950
Predicate DevicesONLINE TDM Vancomycin, K060586
Establishment
Registration1823260, Roche Diagnostics Operations Inc.

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1. DEVICE DESCRIPTION

The ONLINE TDM Vancomycin Gen.3 is a two reagent assay for the in vitro quantitative determination of vancomycin in human serum or plasma on automated clinical chemistry analyzers. It is a homogeneous microparticle agglutination immunoassay based on the kinetic interaction of microparticles in solution (KIMS). A competitive reaction takes place between the drug conjugate and vancomycin in the serum sample for binding to the vancomycin antibody on the microparticles. The resulting kinetic interaction of microparticles is indirectly proportional to the amount of drug present in the sample.

2. INDICATIONS FOR USE

In vitro test for the quantitative determination of vancomycin in serum and plasma on Roche/Hitachi cobas c systems.

A vancomycin test system is a device intended to measure vancomycin, an antibiotic drug, in serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of vancomycin overdose and in monitoring the level of vancomycin to ensure appropriate therapy.

3. TECHNOLOGICAL CHARACTERISTICS

The assay is based on the kinetic interaction of microparticles in a solution (KIMS). Vancomycin antibody is covalently coupled to microparticles and the drug derivative is linked to a macromolecule. The kinetic interaction of microparticles in solutions is induced by binding of drug-conjugate to the antibody on the microparticles and is inhibited by the presence of Vancomycin in the sample. A competitive reaction takes place between the drug conjugate and Vancomycin in the serum sample for binding to the Vancomycin antibody on the microparticles. The resulting kinetic interaction of microparticles is indirectly proportional to the amount of drug present in the sample.

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Reagents - working solutions:

  • . R1 Vancomycin conjugate; piperazine-N,N-bis (2-ethanesulfonic acid) (PIPES) buffer, pH 7.2; preservative; stabilizer
  • R2 Anti-Vancomycin antibody (mouse monoclonal); latex microparticle; 3 -(Nmorpholino) propane sulfonic acid (MOPS) buffer, pH 7.2; stabilizer

4. PREDICATE DEVICE

The predicate device, manufactured by Siemens, which Roche Diagnostics claims substantial equivalence to, is the ONLINE TDM Vancomycin assay. This was cleared in K060586 as a Traditional 510(k) on the Roche/Hitachi 917 and Modular P analyzer systems. The ONLINE TDM Vancomycin assay was then applied to the cobas c 501 analyzer system. FDA designated this CLIA categorization as moderate complexity with the K060373/A001.The following table compares the features of the candidate device to the predicate device.

Assay Comparison
FeaturePredicate Device:
ONLINE TDM Vancomycin
K060586Candidate Device:
ONLINE TDM Vancomycin
Gen.3
Intended UseThe ONLINE TDM Vancomycin
assay is for the quantitative
determination of vancomycin in
human serum or plasma on Roche
automated clinical chemistry
analyzers.In vitro test for the quantitative
determination of vancomycin in
serum and plasma on
Roche/Hitachi cobas c systems.
Analyzer SystemsHitachi / Roche Modular P and 917
cobas c 501 (K060373/A001)cobas c 501
Sample TypesSerum: Collect serum using
standard sampling tubes.
Plasma:Potassium (K2 or K3)
EDTA,sodium citrate, or fluoride
oxalate plasma.Serum
Plasma: K2- or K3-EDTA, lithium
heparin.
Assay Comparison
FeaturePredicate Device:
ONLINE TDM Vancomycin
K060586Candidate Device:
ONLINE TDM Vancomycin
Gen.3
Test PrincipleThe assay is based on a
homogeneous enzyme immunoassay
technique used for the quantitative
analysis of vancomycin in human
serum or plasma. The assay is based
on competition between drug in the
sample and drug labeled with the
enzyme glucose-6-phosphate
dehydrogenase (G6PDH) for
antibody binding sites. Enzyme
activity decreases upon binding to
the antibody, so the drug
concentration in the sample can be
measured in terms of enzyme
activity. Active enzyme converts
oxidized nicotinamide adenine
dinucleotide (NAD) to NADH,
resulting in an absorbance change
that is measured
spectrophotometrically. Endogenous
serum G6PDH does not interfere
because the coenzyme functions
only with the bacterial (Leuconostoc
mesenteroids) enzyme employed in
the assay.The assay is based on the kinetic
interaction of microparticles in a
solution (KIMS). Vancomycin
antibody is covalently coupled to
microparticles and the drug
derivative is linked to a
macromolecule. The kinetic
interaction of microparticles in
solutions is induced by binding of
drug-conjugate to the antibody on
the microparticles and is inhibited
by the presence of Vancomycin in
the sample. A competitive reaction
takes place between the drug
conjugate and Vancomycin in the
serum sample for binding to the
Vancomycin antibody on the
microparticles. The resulting
kinetic interaction of
microparticles is indirectly
proportional to the amount of drug
present in the sample.
Reagent Shelf Life
Stability2-8 °C until expiration dateSame
Reagent On-Board
Stability60 days opened and refrigerated on
the analyzer. Do not freeze.12 weeks on-board in use and
refrigerated on the analyzer. Do
not freeze.
Measuring Range1.7 to 80 µg/mL
(based on LDL)4.0 to 80 µg/mL
(based on LoQ)
TraceabilityThis method has been standardized
against USP reference standards.Same
CalibratorPreciset TDM 1 calibrator
(Previously cleared 510(k):
K031856)Same
Calibration
Frequency• after reagent bottle change
• after reagent lot change
• as required following quality
control procedures• after lot change
• after 6 weeks
• as required following quality
control procedures
ControlsTDM Control Set
(Previously cleared 510(k): K060429
and K070200)Same
Assay Comparison
FeaturePredicate Device:
ONLINE TDM Vancomycin
K060586Candidate Device:
ONLINE TDM Vancomycin
Gen.3
Lower Limits of
MeasurementLower Detection Limit = 1.7
µg/mL (1.2 µmol/L)LoB = 1.0 µg/mL (0.69 µmol/L)
LoD = 1.5 µg/mL (1.04 µmol/L)
LoQ = 4.0 µg/mL (2.76 µmol/L)
Reagent
CompositionR1 Enzyme Reagent
Vancomycin labeled with bacterial
G6PHDH in buffer
R2 Antibody/Substrate Reagent
Anti-vancomycin antibody (mouse
monoclonal), G6P and NAD in
bufferR1 Vancomycin conjugate;
piperazine-N,N'-bis (2
ethanesulfonic acid) (PIPES)
buffer, pH 7.2; preservative;
stabilizer
R2 Anti-Vancomycin antibody
(mouse monoclonal); latex
microparticle; 3-(N-morpholino)
propane sulfonic acid (MOPS)
buffer, pH 7.2; stabilizer

Table 1: Substantial Equivalence – Assav Comparison

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5. NON-CLINICAL PERFORMANCE EVALUATION

The following performance data were provided in support of the substantial equivalence determination:

Detection Limit: LoB, LoD and LoQ according to CLSI EP17-A2

Precision according to CLSI EP5-A2

Linearity according to CLSI EP6-A

Matrix Comparison - Anticoagulants

Interferences - H, L and I Indices

Interference - Drugs

Method Comparison to Predicate

5.1. Detection Limit

LoB, LoD, and LoQ studies were performed based upon CLSI EP17-A2.

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LoB:

LoB (Limit of Blank) corresponds to the concentration below which analyte-free samples are found with a probability of 95%.

For MP Lot: The diluent is measured with 5-fold determinations per run on two instruments. Six runs distributed over 3 days were performed. Data analysis was based on determination of the 95th percentile of the 60 measured values.

For P2 and P3 Lots: The diluent is measured with 5-fold determinations per run on one instrument. Six runs distributed over 3 days were performed. Data analysis was based on determination of the 95th percentile of the 60 measured values.

LoD:

LoD (Limit of Detection) corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

For MP Lot: Five serum samples with low analyte content spiked with Vancomycin (with concentrations ranging from LoB to approx. 4 times specified LoB) were measured with 1-fold determination per run on two instruments. Six runs distributed over 3 days were performed.

For P2 and P3 Lots: Five serum samples with low analyte content spiked with Vancomycin (with concentrations ranging from LoB to approx. 4 times specified LoB) were measured with 2-fold determination per run on one instrument. Six runs distributed over 3 days were performed.

LoQ:

LoQ (Limit of Quantitation) is the lowest amount of analyte in a sample that can be detected quantitatively within specified precision and accuracy ranges.

Fifteen serum samples were prepared which cover the concentration range between LoB and 2x LoQ. Those samples were tested in two aliquots. Six runs over 4 days were performed. Expected value is determined with Vancomycin LCMS/MS.

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| | Result MP Lot
(μg/mL) | Result P2 Lot
(μg/mL) | Result P3 Lot
(μg/mL) | Claim
(μg/mL) |
|-----------------------------|--------------------------|--------------------------|--------------------------|------------------|
| Limit of Blank (LoB) | 0.6 | 0.7 | 0.5 | 1.0 |
| Limit of Detection (LoD) | 1.4 | 1.3 | 1.1 | 1.5 |
| Limit of Quantitation (LoQ) | 2.0 | 3.3 | 3.1 | 4.0 |

Table 2: LoB, LoD, and LoQ Experimental Determination

5.2. Precision according to CLSI EP5-A

Precision experiments were performed in Accordance with CLSI Guideline EP5-A2. Two runs per day for $\geq$ 21 days were performed on the same analyzer. Repeatability and intermediate precision was calculated. The serum samples were randomized in each run separately. The data set was completed for the 21 days. For each sample, the following were calculated: Mean, Repeatability and Intermediate precision as CV and SD values.

SpecimenMean (µg/mL)SD (µg/mL)CV (%)
TDM Control 17.450.45.2
TDM Control 221.50.52.3
TDM Control 336.20.92.4
Human Serum 14.820.48.2
Human Serum 27.950.45.2
Human Serum 332.10.82.5
Human Serum 440.01.02.5
Human Serum 571.42.02.8

Table 3: Repeatability Precision Summary

Table 4: Intermediate Precision Summary

SpecimenMean ( $\mu$ g/mL)SD ( $\mu$ g/mL)CV (%)
TDM Control 17.450.56.2
TDM Control 221.50.83.7
TDM Control 335.51.13.2
Human Serum 14.930.510.5
Human Serum 27.950.55.9
Human Serum 332.11.13.4
Human Serum 439.51.12.9
Human Serum 571.42.23.1

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Linearity according to CLSI EP6-A 5.3.

A dilution series were prepared from a human serum sample pool and diluent (Preciset TDM1Diluent). The dilution series were prepared to obtain sixteen levels* (including the high concentration pool and diluent). The diluted samples shall span the measuring range including a non-zero sample below the measuring range and a sample over the measuring range. The process was repeated for plasma samples.

  • Due to the importance of the trough concentration (ca. 5 – 10 $\mu$g/mL Vancomycin) and due to the unknown LoQ before the measurement, more samples were chosen in the low concentration range.

The calculation is according to the CLSI guideline EP6-A. All measurement data of the dilution steps were calculated by linear regression without weighting.

Sample TypeLinear Regression EquationClaimed Measuring Range
Serumy=1.000x-0.000
Pearson correlation coefficient (R)=0.99854.0 to 80.0 µg/mL
Plasmay=1.000x-0.000
Pearson correlation coefficient (R)=0.99764.0 to 80.0 µg/mL

Table 5: Linearity Results

5.4. Matrix Comparison - Anticoagulants

Each pair of serum and plasma of a single donor are spiked with Vancomycin. Included in the data are 67 full tubes and 9 half-filled tubes (except K2-EDTA plasma, which had 10 tubes). The half-filled and filled sample tubes are also from one donor. Method comparison is executed by taking the serum as reference. Only samples within the measuring range were used.

The following method comparisons are provided:

  • K2-EDTA plasma vs serum ●
  • K3-EDTA plasma vs serum
  • Li-Heparin plasma vs serum ●

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Table 6: Matrix Comparison

AnticoagulantCorrelation
Serum vs. Li-heparin$y = 1.01x -0.3, r = 0.996$
Serum vs. K2-EDTA$y = 0.99x -0.0, r = 0.996$
Serum vs. K3-EDTA$y = 1.00x - 0.3, r = 0.995$

5.5. Interferences - H, L and I Indices

The effect on quantitation of analyte in the presence of endogenous interfering substances is determined at two Vancomycin concentrations and a dilution set of the added interfering substances. Interfering substances evaluated include:

Hemolysis up to an H index of 1000

Lipemia up to an L index of 1000

Icterus/Bilirubin up to an I index of 60

High concentrated stock solutions of the interference substances were prepared in a suitable solvent. Two human serum sample pools were spiked with the defined Vancomvcin concentrations and divided into two aliquots. The potential interfering substance is added to one aliquot, while the other aliquot was mixed with the same amount of solvent without the interfering substance. A dilution series was prepared with 11 dilution steps for each interferent by mixing the 2 aliquots. Three aliquots per level were tested in 1 run on 1 instrument and 1 lot.

The parts containing the interfering substance will have the same Vancomycin concentrations as the aliquots containing no interfering substance. When diluting those two aliquots the Vancomycin concentration will remain constant while the concentration of interferent will vary. Thus the effect of increasing concentrations of interferent can be determined.

Median of the measured results were compared to the expected result (aliquot with no interfering substance) and the recovery is determined (paired difference testing).

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InterferentNo interference up to
HemolysisLevel 1: 1161 H Index
Level 2: 1133 H Index
LipemiaLevel 1: 1234 L Index
Level 2: 1232 L Index
Unconjugated BilirubinLevel 1: 79 I Index
Level 2: 82 I Index
Conjugated BilirubinLevel 1: 77 I Index
Level 2: 75 I Index

Table 7: Interference from Endogenous Substances

Labeling Claim for Endogenous Substances:

Icterus:

No significant interference up to an I index of 60 for conjugated bilirubin and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 60 mg/dL or 1026 umo1/L).

Hemolysis:

No significant interference up to an H index of 1000 (approximate hemoglobin concentration: 1000 mg/dL or 622 umol/L).

Lipemia (Intralipid):

No significant interference up to an L index of 1000. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration. No significant interference from triglycerides up to 1000 mg/dL (11.4 mmol/L).

5.6. Interferences – Drugs

Two human serum sample pools spiked with the defined Vancomycin concentrations were divided into two aliquots. One aliquot of each concentration were used as the reference sample for Vancomycin concentration and were not spiked with the drugs but the solvent for the drug.

The other aliquots, with either the high or low Vancomycin concentration, were spiked with the respective amount of drug. The Vancomycin concentration of the spiked aliquots were tested with 3 replicates in one run, 1 reagent lot and one instrument. The defined pharmaceutical compounds were spiked into samples with concentrations according to EP7-A2 or higher concentrations.

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| Drug | Highest Concentration Shown Not to Interfere with
Vancomycin |
|----------------------|-----------------------------------------------------------------|
| Acetylsalicylic acid | 1000 mg/L |
| Acetaminophen | 200 mg/L |
| Acetylcysteine | 1660 mg/L |
| Ampicillin-sodium | 1000 mg/L |
| Ascorbic acid | 300 mg/L |
| Cefoxitin | 2500 mg/L |
| Cyclosporine | 5 mg/L |
| Doxycycline | 50 mg/L |
| Heparin | 5000 U/L |
| Ibuprofen | 500 mg/L |
| Levodopa | 20 mg/L |
| Methyldopa | 20 mg/L |
| Metronidazole | 200 µg/mL |
| Methotrexate | 455 µg/mL |
| Phenylbutazone | 400 mg/L |
| Rifampicin | 60 mg/L |
| Theophyllin | 100 mg/L |

Table 8: Common Drug Interferences

5.7. Method Comparison to Predicate

One hundred twenty five single native human serum samples of patients taking Vancomycin covering the reportable range were tested. Eight of these native Vancomycin samples were spiked with Vancomycin and 1 sample diluted to cover the range. All samples were tested for icteric, lipemic and hemolytic interference.

The samples were tested in singlicate on the candidate and predicate device (cobas c 501).

The data was evaluated using Passing Bablok Regression analysis.

$$\mathbf{y} = 0.993\mathbf{x} + 0.641, \mathbf{r} = 0.994$$

CONCLUSIONS 6.

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