(104 days)
No
The summary describes a standard immunoassay system with fluorescence detection and a calibration curve. There is no mention of AI or ML in the device description, intended use, or performance studies.
No
The device is an in vitro diagnostic (IVD) device used to quantify 25-Hydroxy vitamin D in human serum and plasma to aid in the assessment of vitamin D sufficiency. It does not directly treat or prevent any condition.
Yes
The device "FREND™ Vitamin D test" is explicitly described as an "in vitro diagnostic use only" test and is used for "quantitative measurement of 25-Hydroxy vitamin D ... to aid in the assessment of vitamin D sufficiency," which are characteristics of a diagnostic device.
No
The device description clearly outlines multiple hardware components including the FREND™ System, FREND™ AP System, FREND™ Vitamin D Cartridge, Vitamin D Pretreatment Tube, and various reagents. While software is mentioned as controlling the user interface and hardware, it is an integral part of a larger hardware-based system for performing an immunoassay.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "For in vitro diagnostic use only."
- Nature of the Test: The device is designed to measure a substance (25-Hydroxy vitamin D and related hydroxylated metabolites) in human biological specimens (serum and plasma). This is a core characteristic of an in vitro diagnostic test.
- Intended Use: The test is used "to aid in the assessment of vitamin D sufficiency," which is a diagnostic purpose.
- Care Setting: The device is intended for use in "clinical laboratories," a typical setting for IVD testing.
The entire description reinforces that this device is used to perform a diagnostic test on samples taken from the human body, outside of the body (in vitro).
N/A
Intended Use / Indications for Use
The FREND™ Vitamin D test is a rapid indirect competitive fluorescent immunoassay designed for the quantitative measurement of 25-Hydroxy vitamin D and related hydroxylated metabolites in human serum and plasma (K;EDTA, lithium-heparin and citrate) specimens using the FREND™ System, and the FREND™ AP System. Measurements of total 25-hydroxy vitamin D and related hydroxylated metabolites are used to aid in the assessment of vitamin D sufficiency.
The FREND™ Vitamin D microfluidic flow cartridge is designed for use in the FREND™ System fluorescent immunoassay reader, and the FREND™ AP System. The FREND™ Vitamin D Test System is intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings.
Product codes (comma separated list FDA assigned to the subject device)
MRG
Device Description
The FREND™ Vitamin D Test is a rapid fluorescence immunoassay designed to quantify the concentration of total 25-hydroxy (OH) vitamin D and related hydroxylated metabolites in human serum and plasma (K3 EDTA, lithium-heparin and citrate) samples using the FREND™ System.
The FREND™ Vitamin D Test System is a competitive immunoassay with gold microparticles labeled with Vitamin D-specific monoclonal anti-Vitamin D-antibody (mouse), Vitamin D-biotin labeled with fluorescence nanoparticles and fluorescence detection by the FREND™ System.
The FREND™ Vitamin D Cartridge is a single-use disposable rapid lateral flow fluorescence immunoassay designed to quantify the concentration of total 25-OH vitamin D and related hydroxylated metabolites in human serum and plasma (K3 EDTA, lithium-heparin and citrate) samples. Each test cartridge contains reagents to perform one test and bears a bar code with test type and lot information. Each FREND™ Vitamin D Cartridge contains a test zone and a reference zone (internal control). The fluorescence measured in the test and reference zones corresponds to the amount of fluorescent conjugates of Vitamin D-biotin that is bound to each zone. Cartridges are provided in individually sealed pouches and must be stored between 2 and 8 °C.
In addition to the QC Code Chip there is a Vitamin D Code Chip. Lot information and calibration data are loaded into the software via the Vitamin D Code Chip provided with each reagent kit. Up to three different Vitamin D Code Chip lots can be loaded and used at the same time. Each reagent cartridge bears a barcode that the FREND™ System uses to identify the test type and lot number and links it to the appropriate analytical program and calibration information. The FREND™ System will not output results if the Vitamin D Code Chip lot does not match the lot of the cartridge used.
The Vitamin D Pretreatment Tube is a small single-use disposable plastic tube that contains Vitamin D gold nano-particle coniugation antibody. The sample is diluted in the dilution tube containing perfluorohexanoic acid and transferred to the pretreatment tube prior to being inserted into the FREND™ AP System to be mixed and then added to the Vitamin D Test Cartridge. The test cartridge is then inserted into the FREND™ System to be analyzed.
The FREND™ AP System is an "Advanced Preparing" device that performs automated pre-analytical steps of mixing, timed heating, and pipetting of the sample from the Gold Antibody Pretreatment tube into the FREND Vitamin D test cartridge.
The FREND™ System utilizes microfluidic technology and detects immune-complexes bound to Vitamin D. The test cartridge is placed on the warming platform of the FREND™ AP System. A 35 µL patient sample is added to the dilution tube and 70 µL of diluted sample is pipetted into the pretreatment tube containing monoclonal anti-Vitamin D antibody conjugated with gold nanoparticles(Step 1). The tube is then loaded into the FREND™ AP System (Step 2). The tube is mixed and loaded into the test cartridge automatically. In the AP System (Step 3), mixing, reaction, sample addition, hydration of the Vitamin D-biotin fluorescent bead conjugate, and migration along the cartridge channel takes approximately 15 minutes, after which the cartridge is read on the FREND™ System. During migration, the bound Vitamin D in the sample and the fluorescent bead conjugates of Vitamin D-biotin compete to form antigen-antibody complex in the test zone. Unbound Vitamin D-biotin fluorescent conjugates flow through and bind to the anti-Vitamin D antibody that is fixed on the surface in the reference zone. The fluorescent signals in the test and reference zones are measured, typically within 4 minutes.
Vitamin D quantification is based upon the ratio of the intensity of the test and reference zones. A lower ratio of fluorescence is indicative of a higher Vitamin D concentration; in other words, the magnitude of the fluorescent ratio is inversely proportional to the amount of Vitamin D in the sample.
The measuring range of the FREND™ Vitamin D Test System is 13.0 to 96.0 ng/mL. Results are determined via a lot-specific calibration curve which is generated by the manufacturer using a four-point calibration determined from values averaged from 6 replicates at each level. The established curve is uploaded to the FREND™ System via the Vitamin D Code-chip and is valid until the lot expiration date.
The FREND™ System is a portable, automated FREND™ cartridge reader. The FREND™ System is based on quantitative immunoassay technology capable of quantifying single or multiple analytes by measuring laser-induced fluorescence in a single-use disposable reagent cartridge. The test cartridge is a disposable plastic device that contains a port or opening (inlet) where the sample is applied. Once the sample is applied, it will mix with the reagents and travel towards the detection area via capillary action.
The FREND™ cartridge utilizes microfluidics lateral flow technology where the analyte of interest in the sample forms immune complexes while moving through the fluidics pathway in the cartridge. The concentration of the analyte of interest in an unknown sample is calculated using the ratio of the fluorescent intensity of the test zone and the reference zone.
The FREND™ System is a bench top fluorescence reader containing a touch screen user interface. The System has a slot that accepts the FREND™ Vitamin D Test Cartridge (which contains the reagents and sample), and is programmed to analyze the Test when the sample has fully reacted with the on-board in-cartridge reagents. Results of the test are displayed on the screen and can be printed on an optional printer.
The FREND™ System software controls the graphical user interface, communication with hardware, database management and data analysis. The software also controls the functions of the mechanical components including the motor, laser, printer control and acquisition of data from the sensor. The user can set the time and date and enter patient ID through the graphic user interface. The user cannot make any changes to the software.
The FREND™ Vitamin D Test System includes the following in the reagent kit:
- . 20 FREND™ Vitamin D Test Cartridges
- 20 FREND™ Sample Dilution Tubes .
- 20 FREND™ Vitamin D Gold Antibody Pretreatment Tubes .
- 30 Disposable pipette tips
- 1 FREND™ Vitamin D Code Chip .
- . 1 FREND™ Vitamin D Package Insert
One cartridge contains:
- . Monoclonal mouse anti- Vitamin D 1.6 ± 0.16 na
- . 25-hydroxyvitamin D 136 ± 13.6 ng
- . Fluorescence particles 2.4 ± 0.24 µg
One sample dilution tube contains:
- . Perfluorohexanoic acid 9.5 ± 0.95 µg
One Gold antibody pretreatment tube contains: - . Gold nano-particle conjugation antibody 7.0 ± 0.7 µg
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings.
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision/Reproducibility: A single lot imprecision study was performed at the NanoEnTek laboratory as described in the CLSI protocol EP5-A3. Three serum pools with low, intermediate and high Vitamin D levels were assayed in duplicate twice per day for 20 days (80 total measurements per level).
Results: Sample 1 (mean 20.1 ng/mL): Repeatability SD 1.76, CV% 8.8%; Between-run SD 0.46, CV% 2.3%; Between-day SD 0.22, CV% 1.1%; Within-laboratory SD 1.83, CV% 9.1%. Sample 2 (mean 50.6 ng/mL): Repeatability SD 2.26, CV% 4.5%; Between-run SD 1.31, CV% 2.6%; Between-day SD 0.59, CV% 1.2%; Within-laboratory SD 2.67, CV% 5.3%. Sample 3 (mean 80.5 ng/mL): Repeatability SD 4.53, CV% 5.6%; Between-run SD 1.13, CV% 1.4%; Between-day SD 0.74, CV% 0.9%; Within-laboratory SD 4.72, CV% 5.9%.
Linearity/assay reportable range: Linearity was established according to CLSI-EP6-A using nine levels of serum Vitamin D tested in quadruplicate. Linearity was demonstrated across the measuring range of the FREND™ Vitamin D (13.0 ng/mL ~ 96.0 ng/mL).
Traceability, Stability, Expected values (controls, calibrators, or methods): The calibrators are for factory calibration. The standards/calibrators are prepared gravimetrically and confirmed by measurement on the ARCHITECT i 25-OH Vitamin D assay (K110619). All calibration statistics and information have been electronically stored on the FREND™ Vitamin D Code chip included in each box of FREND™ Vitamin D cartridges. There is no need for calibration by the operator as the calibration information is in each code chip, which is lot-specific.
Stability: Real-time stability testing for the Vitamin D reagent kit was performed according to CLSI EP25-A. Evaluation of Stability of In Vitro Diagnostic Reagents. Reagent stability studies based on procedures and criteria in the NanoEnTek quality system showed that the cartridges for FREND™ Vitamin D will meet performance acceptance criteria for one year from the date of manufacturer if stored refrigerated appropriately as directed.
Detection Limit: The Limit of Quantitation (LoQ) for the FREND™ Vitamin D Test System was established at 9.9 ng/mL according to the CLSI EP17-A2 protocol. The analytical sensitivity of the FREND™ Vitamin D Test System is claimed at 13.0 ng/mL.
Analytical specificity (Interference Studies): Interference was determined according to CLSI EP07-A2 using triplicate determinations and two concentration of Vitamin D. No interference was specified if recoveries were between 90% to 110% of the expected Vitamin D value.
Results: Hemoglobin (500 mg/dL): 107.0% (low), 98.4% (high); Bilirubin, conjugated (30 mg/dL): 95.6% (low), 95.7% (high); Bilirubin, unconjugated (30 mg/dL): 100.0% (low), 94.9% (high); Triglyceride (1500 mg/dL): 99.6% (low), 94.0% (high); Cholesterol (500 mg/dL): 93.0% (low), 96.1% (high); Total protein (12 g/dL): 103.4% (low), 98.8% (high); Biotin (6 µg/mL): 99.9% (low), 102.7% (high); HAMA (1000 ng/mL): 97.4% (low), 108.2% (high); Rheumatoid Factor (536 IU/mL): 101.4% (low), 96.8% (high).
Analytical specificity (Cross Reactivity): The following substances were evaluated in triplicate for potential cross-reactivity with the FREND™ Vitamin D Test System at two Vitamin D concentrations. Testing was done according to the CLSI protocol EP07-A2.
Results: No significant cross-reactivity was found, with the exception of Paricalcitol. Drugs containing Paricalcitol (e.g., Zemplar) will interfere with the assay. Cross-reactivity was measured for Vitamin D2 (Ergocalciferol) (0.5-0.8%), Vitamin D3 (Cholecalciferol) (0.5-0.7%), 1,25-(OH)2-Vitamin D2 (2.2-0.7%), 1,25-(OH)2-Vitamin D3 (Calcitriol) (6.9-1.9%), 3-epi-25-(OH) Vitamin D3 (2.1-2.0%), 25-(OH) Vitamin D2 (97.2-97.3%), 25-(OH) Vitamin D3 (107.3-92.2%), and Paricalcitol (15.8-14.7%).
Method comparison with predicate device: Comparison studies with 133 de-identified leftover samples were performed in a CLIA-certified hospital laboratory. The reference method was the Abbott ARCHITECT i 25-OH Vitamin D Assay (K110619) run on the Abbott ARCHITECT i System. The samples spanned the measuring range of the FREND Vitamin D Test System. All samples were assayed using sera split between the applicant device and the reference method.
Results: Results from the FREND™ Vitamin D on the FREND™ System (y) were compared with the reference results (x) by Passing-Bablok regression analysis, giving a slope of 1.069, a y-intercept of -0.182, and a correlation (R) of 0.971.
Matrix comparison: The matrix comparison study was performed at the NanoEnTek laboratory according to CLSI EP14-A3. Vitamin D concentrations from 36 paired samples, covering the analytical measuring range, were measured in serum, K3EDTA, lithium-heparin and sodium citrate plasma samples with the FREND™ Vitamin D Test System, giving equivalent results.
Expected values/Reference range: A study was conducted to establish a Reference Interval for the FREND™ Vitamin D Test System. A total of 300 serum specimens from apparently healthy individuals residing in north, central and southern regions of the US were collected during summer and winter months and tested in the FREND™ Vitamin D Test System.
Reference Interval:
§ 862.1825 Vitamin D test system.
(a)
Identification. A vitamin D test system is a device intended for use in clinical laboratories for the quantitative determination of 25-hydroxyvitamin D (25-OH-D) and other hydroxylated metabolites of vitamin D in serum or plasma to be used in the assessment of vitamin D sufficiency.(b)
Classification. Class II (special controls). Vitamin D test systems must comply with the following special controls:(1) Labeling in conformance with 21 CFR 809.10 and
(2) Compliance with existing standards of the National Committee on Clinical Laboratory Standards.
0
Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo consists of a stylized image of an eagle with three heads in profile, facing right. The eagle is enclosed in a circle with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter of the circle.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
NANOENTEK USA, INC. C/O MAŬREEN GARNER NEW WORLD REGULATORY SOLUTIONS, INC 1983 HAZELWOOD ROAD TOMS RIVER NJ 08753
January 12, 2017
Re: K162754
Trade/Device Name: FREND™ Vitamin D Test System Regulation Number: 21 CFR 862.1825 Regulation Name: Vitamin D test system Regulatory Class: II Product Code: MRG Dated: November 23, 2016 Received: November 28, 2016
Dear Maureen Garner:
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.
Katherine Serrano -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
Indications for Use
510(k) Number (if known)
Device Name FREND™ Vitamin D Test System
Indications for Use (Describe)
The FREND™ Vitamin D test is a rapid indirect competitive fluorescent immunoassay designed for the quantitative measurement of 25-Hydroxy vitamin D and related hydroxylated metabolites in human serum and plasma (K;EDTA, lithium-heparin and citrate) specimens using the FREND™ System, and the FREND™ AP System. Measurements of total 25-hydroxy vitamin D and related hydroxylated metabolites are used to aid in the assessment of vitamin D sufficiency.
The FREND™ Vitamin D microfluidic flow cartridge is designed for use in the FREND™ System fluorescent immunoassay reader, and the FREND™ AP System. The FREND™ Vitamin D Test System is intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings.
X 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."
3
510(k) Summary
As required by the Safe Medical Devices Act (SMDA) of 1990 and in accordance with 21 CFR §807.92, a 510(k) summary is provided for K162754.
A. Applicant
| Company Name: | NanoEnTek, Inc. |
|---|---|
| Address: | 12F 5 Digital-ro, 26-gil, Guro-gu |
| Seoul 152-740, KOREA | |
| Contact Person: | Sunmi Han |
| Phone Number: | +82-2-6220-7940 |
| Facsimile Number: | +82-2-6220-7721 |
B. 510(k) Preparer Information (Contact Person)
| Company Name: | New World Regulatory Solutions, Inc |
|---|---|
| Address: | P.O. Box 5374 |
| Toms River, New Jersey 08754, USA | |
| Contact Person: | Maureen Garner |
| Phone Number: | (732) 779-7422 |
| Facsimile Number: | (732) 270-4829 |
| Email: | NWRSinc@gmail.com |
Date Prepared:
January 6, 2017
C. Purpose for Submission:
New Analyte on FREND™ System
D. Measurand:
Vitamin D
E. Type of Test:
Quantitative, Fluorescence Immunoassay
F. Proprietary and Established Device Name:
FREND™ Vitamin D Test System
Regulatory Information: G.
| Test Kit | |
|---|---|
| Classification Name: | Vitamin D Test System |
| Classification: | Class II |
| Generic Name: | Competitive Immunoassay, Vitamin D |
| Proprietary Name: | FRENDTM Vitamin D |
| (reagent, cartridge assay) | |
| Regulation Number: | 21 CFR §862.1825 |
| Product Code: | MRG |
| Panel | Clinical Chemistry (75) |
Confidential and Proprietary Copyright © 2016 NanoEnTek
4
H. Intended Use:
-
- Intended use(s): See indications for use below:
-
- Indication(s) for use:
The FREND™ Vitamin D Test System is a rapid indirect competitive fluorescent immunoassay designed for the quantitative measurement of 25-hydroxy vitamin D and related hydroxylated metabolites in human serum and plasma (KsEDTA, lithiumheparin and citrate) specimens using the FREND™ System, and the FREND™ AP System. Measurements of total 25-hydroxy vitamin D and related hydroxylated metabolites are used to aid in the assessment of vitamin D sufficiency.
The FREND™ Vitamin D microfluidic flow cartridge is designed for use in the FREND™ System fluorescent immunoassay reader, and the FREND™ AP System. The FREND™ Vitamin D Test System is intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings.
-
- Special conditions for use statement(s): For prescription use only.
-
- Special instrument requirements: NanoEnTek FREND™ System and FREND™ AP System
l. Device Description
The FREND™ Vitamin D Test is a rapid fluorescence immunoassay designed to quantify the concentration of total 25-hydroxy (OH) vitamin D and related hydroxylated metabolites in human serum and plasma (K3 EDTA, lithium-heparin and citrate) samples using the FREND™ System.
The FREND™ Vitamin D Test System is a competitive immunoassay with gold microparticles labeled with Vitamin D-specific monoclonal anti-Vitamin D-antibody (mouse), Vitamin D-biotin labeled with fluorescence nanoparticles and fluorescence detection by the FREND™ System.
The FREND™ Vitamin D Cartridge is a single-use disposable rapid lateral flow fluorescence immunoassay designed to quantify the concentration of total 25-OH vitamin D and related hydroxylated metabolites in human serum and plasma (K3 EDTA, lithiumheparin and citrate) samples. Each test cartridge contains reagents to perform one test and bears a bar code with test type and lot information. Each FREND™ Vitamin D Cartridge contains a test zone and a reference zone (internal control). The fluorescence measured in the test and reference zones corresponds to the amount of fluorescent conjugates of Vitamin D-biotin that is bound to each zone. Cartridges are provided in individually sealed pouches and must be stored between 2 and 8 °C.
In addition to the QC Code Chip there is a Vitamin D Code Chip. Lot information and calibration data are loaded into the software via the Vitamin D Code Chip provided with each reagent kit. Up to three different Vitamin D Code Chip lots can be loaded and used at the same time. Each reagent cartridge bears a barcode that the FREND™ System uses to identify the test type and lot number and links it to the appropriate analytical
Confidential and Proprietary Copyright © 2016 NanoEnTek
5
program and calibration information. The FREND™ System will not output results if the Vitamin D Code Chip lot does not match the lot of the cartridge used.
The Vitamin D Pretreatment Tube is a small single-use disposable plastic tube that contains Vitamin D gold nano-particle coniugation antibody. The sample is diluted in the dilution tube containing perfluorohexanoic acid and transferred to the pretreatment tube prior to being inserted into the FREND™ AP System to be mixed and then added to the Vitamin D Test Cartridge. The test cartridge is then inserted into the FREND™ System to be analyzed.
The FREND™ AP System is an "Advanced Preparing" device that performs automated pre-analytical steps of mixing, timed heating, and pipetting of the sample from the Gold Antibody Pretreatment tube into the FREND Vitamin D test cartridge.
The FREND™ System utilizes microfluidic technology and detects immune-complexes bound to Vitamin D. The test cartridge is placed on the warming platform of the FREND™ AP System. A 35 µL patient sample is added to the dilution tube and 70 µL of diluted sample is pipetted into the pretreatment tube containing monoclonal anti-Vitamin D antibody conjugated with gold nanoparticles(Step 1). The tube is then loaded into the FREND™ AP System (Step 2). The tube is mixed and loaded into the test cartridge automatically. In the AP System (Step 3), mixing, reaction, sample addition, hydration of the Vitamin D-biotin fluorescent bead conjugate, and migration along the cartridge channel takes approximately 15 minutes, after which the cartridge is read on the FREND™ System. During migration, the bound Vitamin D in the sample and the fluorescent bead conjugates of Vitamin D-biotin compete to form antigen-antibody complex in the test zone. Unbound Vitamin D-biotin fluorescent conjugates flow through and bind to the anti-Vitamin D antibody that is fixed on the surface in the reference zone. The fluorescent signals in the test and reference zones are measured, typically within 4 minutes.
Vitamin D quantification is based upon the ratio of the intensity of the test and reference zones. A lower ratio of fluorescence is indicative of a higher Vitamin D concentration; in other words, the magnitude of the fluorescent ratio is inversely proportional to the amount of Vitamin D in the sample.
The measuring range of the FREND™ Vitamin D Test System is 13.0 to 96.0 ng/mL. Results are determined via a lot-specific calibration curve which is generated by the manufacturer using a four-point calibration determined from values averaged from 6 replicates at each level. The established curve is uploaded to the FREND™ System via the Vitamin D Code-chip and is valid until the lot expiration date.
The FREND™ System is a portable, automated FREND™ cartridge reader. The FREND™ System is based on quantitative immunoassay technology capable of quantifying single or multiple analytes by measuring laser-induced fluorescence in a single-use disposable reagent cartridge. The test cartridge is a disposable plastic device that contains a port or opening (inlet) where the sample is applied. Once the sample is applied, it will mix with the reagents and travel towards the detection area via capillary action.
The FREND™ cartridge utilizes microfluidics lateral flow technology where the analyte of interest in the sample forms immune complexes while moving through the fluidics Confidential and Proprietary Copyright © 2016 NanoEnTek
6
pathway in the cartridge. The concentration of the analyte of interest in an unknown sample is calculated using the ratio of the fluorescent intensity of the test zone and the reference zone.
The FREND™ System is a bench top fluorescence reader containing a touch screen user interface. The System has a slot that accepts the FREND™ Vitamin D Test Cartridge (which contains the reagents and sample), and is programmed to analyze the Test when the sample has fully reacted with the on-board in-cartridge reagents. Results of the test are displayed on the screen and can be printed on an optional printer.
The FREND™ System software controls the graphical user interface, communication with hardware, database management and data analysis. The software also controls the functions of the mechanical components including the motor, laser, printer control and acquisition of data from the sensor. The user can set the time and date and enter patient ID through the graphic user interface. The user cannot make any changes to the software.
The FREND™ Vitamin D Test System includes the following in the reagent kit:
- . 20 FREND™ Vitamin D Test Cartridges
- 20 FREND™ Sample Dilution Tubes .
- 20 FREND™ Vitamin D Gold Antibody Pretreatment Tubes .
- 30 Disposable pipette tips
- 1 FREND™ Vitamin D Code Chip .
- . 1 FREND™ Vitamin D Package Insert
One cartridge contains:
- . Monoclonal mouse anti- Vitamin D 1.6 ± 0.16 na
- . 25-hydroxyvitamin D 136 ± 13.6 ng
- . Fluorescence particles 2.4 ± 0.24 µg
One sample dilution tube contains:
-
. Perfluorohexanoic acid 9.5 ± 0.95 µg
One Gold antibody pretreatment tube contains: -
. Gold nano-particle conjugation antibody 7.0 ± 0.7 µg
The FREND™ System (previously cleared in K124056 (FREND™ PSA), K131928 (FREND™ TSH), K152422 (FREND™ Free T4), and K153577 (FREND™ Testosterone) is not provided with the kit but it is required for the FREND™ Vitamin D Test cartridge.
7
Substantial Equivalence Information: J.
An overview of the similarities and differences between the FREND™ Vitamin D Test System and the predicate is provided in the table below:
| Similarities | ||
|---|---|---|
| Item | FRENDTM Vitamin D Test System | Abbott ARCHITECT i 25-OH Vitamin D Assay (K110619) |
| Intended Use | The FRENDTM Vitamin D test is a rapid indirect competitive fluorescent immunoassay designed for the quantitative measurement of 25-hydroxy vitamin D and related hydroxylated metabolites in human serum and plasma (K3 EDTA, lithium-heparin and citrate) specimens using the FRENDTM System, and the FRENDTM AP System. Measurements of total 25-hydroxy vitamin D and related hydroxylated metabolites are used to aid in the assessment of Vitamin D sufficiency. |
The FRENDTM Vitamin D microfluidic flow cartridge is designed for use in the FRENDTM System fluorescent immunoassay reader. and the FRENDTM AP System. The FRENDTM Vitamin D Test System is intended for use in clinical laboratories. For in vitro diagnostic use only. The test is not intended for use in point-of-care settings. | The ARCHITECT 25-OH Vitamin D assay is a chemiluminescent microparticle immunoassay (CMIA) for the quantitative determination of 25-hydroxy vitamin D (25-OH Vitamin D) in human serum and plasma. The ARCHITECT 25-OH Vitamin D assay is to be used as an aid in the assessment of Vitamin D sufficiency.
The ARCHITECT 25-OH Vitamin D Calibrators are for the calibration of the ARCHITECT i System when used for the quantitative determination of 25-hydroxy Vitamin D (25-OH Vitamin D) in human serum and plasma.
The ARCHITECT 25-OH Vitamin D Controls are for the estimation of test precision and the detection of systematic analytical deviations of the ARCHITECT i System when used for the quantitative determination of 25-hydroxy Vitamin D (25-OH Vitamin D) in human serum and plasma.
For in vitro diagnostic use only. |
| Sample Type | Human serum and K3EDTA, lithium heparin, and citrate plasma | Human serum, lithium heparin and sodium heparin |
| Analyte | 25-OH Vitamin D | Same |
| Type of Test | Fluorescent immunoassay determination of Vitamin D | Chemiluminescent immunoassay determination of Vitamin D |
| Quality Control | Internal procedural/instrument quality controls; commercially-available external positive and negative assay controls | Internal procedural/instrument quality controls; external positive and negative assay controls cleared with test kit |
| Interpretation of Results | Inversely proportional Interpolation of fluorescence against a standard curve | Same |
| Dynamic Range | 13.0 ng/mL to 96.0 ng/mL | Same |
8
| Differences | ||
|---|---|---|
| Item | FREND™ Vitamin D | |
| Test System | Abbott ARCHITECT i 25-OH Vitamin D | |
| Assay (K110619) | ||
| Sample Size | 35 µL for the dilution step and 70 µL for | |
| running the test | 60 µL (priority) or 150 µL (≤3h) for the | |
| first test plus 10 µL for each additional | ||
| test from the same test cup | ||
| Test Cartridge | Disposable single-use cartridge | No single-use cartridge |
| Random | ||
| Access / | ||
| Degree of | ||
| Automation | No random access, manual | |
| manipulation | Random access, semi-automated |
K. Performance Characteristics (if/when applicable)
-
- Analytical performance
a. Precision/Reproducibility
A single lot imprecision study was performed at the NanoEnTek laboratory as described in the CLSI protocol EP5-A3. Three serum pools with low, intermediate and high Vitamin D levels were assaved in duplicate twice per day for 20 days (80 total measurements per level). The results are summarized below:
| Sample
Pool | Mean Vitamin D
Concentration
(ng/mL) | Repeatability | | Between-
run | | Between-
day | | Within-
laboratory | |
|----------------|--------------------------------------------|---------------|------|-----------------|------|-----------------|------|-----------------------|------|
| | | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| 1 | 20.1 | 1.76 | 8.8% | 0.46 | 2.3% | 0.22 | 1.1% | 1.83 | 9.1% |
| 2 | 50. 6 | 2.26 | 4.5% | 1.31 | 2.6% | 0.59 | 1.2% | 2.67 | 5.3% |
| 3 | 80.5 | 4.53 | 5.6% | 1.13 | 1.4% | 0.74 | 0.9% | 4.72 | 5.9% |
| FREND™ Vitamin D Single Site Single Lot Precision |
|---|
- b. Linearity/assay reportable range:
Linearity was established according to CLSI-EP6-A using nine levels of serum Vitamin D tested in quadruplicate. Linearity was demonstrated across the measuring range of the FREND™ Vitamin D (13.0 ng/mL ~ 96.0 ng/mL).
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
The calibrators are for factory calibration. The standards/calibrators are prepared gravimetrically and confirmed by measurement on the ARCHITECT i 25-OH Vitamin D assay (K110619). All calibration statistics and information have been electronically stored on the FREND™ Vitamin D Code chip included in each box of FREND™ Vitamin D cartridges. There is no need for calibration by the operator as the calibration information is in each code chip, which is lot-specific.
- d. Stability
Real-time stability testing for the Vitamin D reagent kit was performed according to CLSI EP25-A. Evaluation of Stability of In Vitro Diagnostic Reagents. Reagent stability studies based on procedures and criteria in the NanoEnTek quality system showed that the cartridges for FREND™ Vitamin D will meet performance acceptance criteria for one year from the date of manufacturer if stored refrigerated appropriately as directed.
Confidential and Proprietary Copyright © 2016 NanoEnTek
9
-
e. Detection Limit:
The Limit of Quantitation (LoQ) for the FREND™ Vitamin D Test System was established at 9.9 ng/mL according to the CLSI EP17-A2 protocol. The analytical sensitivity of the FREND™ Vitamin D Test System is claimed at 13.0 ng/mL. -
f. Analytical specificity:
Interference Studies
Interference was determined according to CLSI EP07-A2 using triplicate determinations and two concentration of Vitamin D. No interference was specified if recoveries were between 90% to 110% of the expected Vitamin D value. Results are summarized in the table below.
| Class | Interferent (Concentration Tested) | % Recovery
Vitamin D
Low | % Recovery
Vitamin D
High |
|----------------------------|------------------------------------|--------------------------------|---------------------------------|
| Endogenous
substances | Hemoglobin (500 mg/dL) | 107.0 | 98.4 |
| | Bilirubin, conjugated (30 mg/dL) | 95.6 | 95.7 |
| | Bilirubin, unconjugated (30 mg/dL) | 100.0 | 94.9 |
| | Triglyceride (1500 mg/dL) | 99.6 | 94.0 |
| | Cholesterol (500 mg/dL) | 93.0 | 96.1 |
| | Total protein (12 g/dL) | 103.4 | 98.8 |
| | Biotin (6 µg/mL) | 99.9 | 102.7 |
| Heterophilic
Antibodies | HAMA (1000 ng/mL) | 97.4 | 108.2 |
| | Rheumatoid Factor (536 IU/mL) | 101.4 | 96.8 |
Cross Reactivity
The following substances were evaluated in triplicate for potential cross-reactivity with the FREND™ Vitamin D Test System at two Vitamin D concentrations. Testing was done according to the CLSI protocol EP07-A2. No significant cross-reactivity was found, with the exception of Paricalcitol. Drugs containing Paricalcitol (e.g., Zemplar) will interfere with the assay.
| Cross-reactant | Cross-reactant Concentration (ng/mL) | % Cross-reactivity Vitamin D Low | % Cross-reactivity Vitamin D High |
|---|---|---|---|
| Vitamin D2 (Ergocalciferol) | 500 | 0.5 | 0.8 |
| Vitamin D3 (Cholecalciferol) | 500 | 0.5 | 0.7 |
| 1,25-(OH)2-Vitamin D2 | 100 | 2.2 | 0.7 |
| 1,25-(OH)2-Vitamin D3 (Calcitriol) | 100 | 6.9 | 1.9 |
| 3-epi-25-(OH) Vitamin D3 | 100 | 2.1 | 2.0 |
| 25-(OH) Vitamin D2 | 25 | 97.2 | 97.3 |
| 25-(OH) Vitamin D3 | 25 | 107.3 | 92.2 |
| Paricalcitol | 25 | 15.8 | 14.7 |
% Cross-reactivity = 100 x ((Measured value - true value)/concentration of interferent)
g. Assay cut-off:
Not applicable
10
2. Comparison studies
- a. Method comparison with predicate device:
Comparison studies with 133 de-identified leftover samples were performed in a CLIA-certified hospital laboratory. The reference method was the Abbott ARCHITECT i 25-OH Vitamin D Assay (K110619) run on the Abbott ARCHITECT i System. The samples spanned the measuring range of the FREND Vitamin D Test System. All samples were assayed using sera split between the applicant device and the reference method.
Results from the FREND™ Vitamin D on the FREND™ System (y) were compared with the reference results (x) by Passing-Bablok regression analysis, giving a slope of 1.069, a y-intercept of -0.182, and a correlation (R) of 0.971.
- b. Matrix comparison:
The matrix comparison study was performed at the NanoEnTek laboratory according to CLSI EP14-A3. Vitamin D concentrations from 36 paired samples, covering the analytical measuring range, were measured in serum, K3EDTA, lithium-heparin and sodium citrate plasma samples with the FREND™ Vitamin D Test System, giving equivalent results.
3. Expected values/Reference range:
A study was conducted to establish a Reference Interval for the FREND™ Vitamin D Test System. It is also recommended that each laboratory establish its own reference range, which may be unique to the population it serves depending upon geographical season, patient, dietary, or environmental factors.
The study was conducted based on CLSI C28-A3. A total of 300 serum specimens from apparently healthy individuals residing in north, central and southern regions of the US were collected during summer and winter months and tested in the FREND™ Vitamin D Test System. The reference interval is provided below.
| Reference Interval | Lower reference limit
90% CI | Upper reference limit
90% CI |
|--------------------|---------------------------------|---------------------------------|
|