The FREND™ PSA Plus as performed on the FREND™ system, is a quantitative in vitro diagnostic test which measures total Prostate Specific Antigen (PSA) in human serum and plasma. The NanoEnTek FREND™ PSA Plus is designed for in vitro DIAGNOSTIC USE ONL Y for the quantitative measurement of total Prostate Specific Antigen (PSA) in human serum, heparinized plasma, and EDTA plasma using the FREND™ System. This device is indicated for the serial measurement of total PSA in serum, heparinized plasma and EDTA plasma to be used as an aid in the management of patients with prostate cancer.

The FREND™ PSA Plus is indicated for use in clinical laboratories upon prescription by the attending physician as an aid to clinicians in managing patients with prostate cancer.

The information provided from this test may supplement decision-making and should only be used in conjunction with routine monitoring by a physician and the use of other diagnostic procedures. Because of the variability in the effects of various medications used in the treatment of prostate cancer, clinicians should use professional judgment in the interpretation of PSA results as an indicator of disease status.

Device Description

The FREND™ PSA Plus is a rapid fluorescence immunoassay that measures prostate specific antigen (PSA) in human serum and in lithium heparin and EDTA plasma using the FREND™ system. The FREND™ PSA Plus is intended for use as an aid for prostate cancer management. The FREND™ PSA Plus Test is a single use fluorescence immunoassay designed to quantify the concentration of total PSA in serum and lithium heparin and EDTA plasma samples. The specimen is added by the operator to the sample inlet with a transfer pipet, allowing the appropriate volume of sample (30 µL) to be delivered into the FREND™ PSA Plus Test Cartridge. The Cartridge is then placed into the FREND™ System, which is programmed to begin analysis once the sample has reacted with the reagents. The reaction and analysis time is approximately 6 minutes. The PSA quantification is based on the amount of fluorescence detected by the FREND™ System at the FREND™ PSA Plus Test Cartridge window. A higher level of fluorescence is indicative of a higher PSA concentration. In other words, the magnitude of the fluorescent signal is directly proportional to the amount of total PSA in the sample.

The FREND™ System is a bench top fluorescence reader containing a touchscreen user interface. The System has a slot that accepts the FREND™ PSA Plus 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 through the RS232C interface.

AI/ML Overview

Here's a summary of the acceptance criteria and study findings for the FREND™ PSA Plus on the FREND™ system, based on the provided 510(k) summary:

1. Table of Acceptance Criteria and Reported Device Performance

| Performance Metric | Acceptance Criteria (Stated or Implied) | Total (Total CV%) | |
| Site-to-Site Value) | 3.50% | 1.57% | 1.67% | 3.47% | 1.61% | 2.06% | |
| Inter-cartridge | 18.45% | 6.81% | 7.94% | 20.03% | 6.17% | 7.49% | |
| Total (Total CV%) | 20.87% | 7.74% | 10.79% | 21.22% | 7.69% | 9.81% | |

Note: The document explicitly states "acceptance criteria" for some tests (e.g., dilution linearity, spiked recovery, interference) but for others (e.g., imprecision, method comparison), it describes the results and concludes they are "acceptable" or "compared well," implying that the performance met internal thresholds comparable to predicate devices.

2. Sample Sizes and Data Provenance

Clinical Samples: 1219 evaluable clinical serum samples.
- Provenance: Prospectively collected stored samples were utilized for the clinical study. No specific country of origin is explicitly stated, but NanoEnTek is a Korean company with a CRO (DOCRO, Inc.) in the US, and testing was done at both NanoEnTek facilities and CLIA licensed facilities in the US.
Precision (Analytical):
- Intra-assay/inter-assay/complex imprecision: Three clinical samples (0.186, 2.757, 16.625 ng/mL) assayed in duplicates twice a day for 20 days using a single lot cartridge (total 80 measurements per sample).
- Multi-Site, Multi-Lot Imprecision: Four replicates each of Material A, B, C and two replicates of QC 1, 2, 3 were evaluated in two runs performed for five days at each of three geographically diverse sites. This yielded a total of 40 results on each material per site, and 120 total replicates for each material across all sites.
Dilution Linearity & Recovery: A serum pool with elevated PSA (34 ng/mL) was diluted to seven levels, plus neat and zero samples. Each level was tested in 6 replicates.
Spiked Recovery: A serum pool from females (tPSA < 0.01 ng/mL) was spiked with 3 known PSA levels. Samples were assayed before and after spiking (number of replicates not explicitly stated, but table shows 3 replicates per spike level).
Analytical Sensitivity (LOD/LOQ): 5 blank samples and 5 low PSA concentration samples were tested in 12 replicates each.
High Dose Hook Effect: A concentrated sample of purified PSA antigen tested neat and on dilution (number of replicates not specified).
Interfering Substances and Assay Specificity: Not explicitly stated, but samples were tested according to CLSI protocol EP7-A.
Method Comparison:
- Prostate cancer samples: Single point samples (n = 85) and earliest sample from serially monitored subjects (n = 75), for a total of 160 samples (tPSA < 25 ng/mL).
- Serially Monitored Subjects: 75 subjects undergoing serial monitoring for prostate cancer. A total of 236 point-to-point determinations for each assay.
Sample Matrix Comparison Study: 36 matched sets (serum, lithium heparin, EDTA plasma, sodium citrate) were collected, aliquoted, frozen, and run.
Reagent Stability Studies: Three different lots of cartridges were tested over 15 months at three-month intervals using five standard specimens.

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

The document does not explicitly state the number of experts or their specific qualifications (e.g., radiologist with X years of experience) for establishing the ground truth for the test set in the clinical performance section.

For the serial monitoring study, "Disease status for the patients was determined by the physician. This Disease Status was used to determine Progression or No Progression from a clinical perspective." This implies a medical professional established the clinical ground truth. The nature of this determination included "other laboratory tests, patient interviews, physical examinations, and imaging studies of a variety of types."

4. Adjudication Method for the Test Set

The document does not describe a formal adjudication method (like 2+1 or 3+1 consensus) for the clinical test set. The clinical status for serially monitored patients was determined by "the physician" and was used as the ground truth.

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

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not conducted. This device is an in vitro diagnostic assay, not an imaging device or AI-assisted diagnostic tool that would typically involve human readers interpreting cases with and without AI assistance. The clinical study compares the device's performance to a predicate device and clinical status, but not the improvement of human reader performance.

6. Standalone (Algorithm Only) Performance

Yes, numerous standalone performance studies were done. The FREND™ PSA Plus is an automated immunoassay system, and its analytical and clinical performance is its standalone performance. The entire "Performance Data Summary - Analytical Testing" and "Performance Data Summary - Clinical Testing" sections detail the standalone performance of the assay and system without human intervention in the measurement process (though human input is required to operate the system and interpret results in a clinical context).

7. Type of Ground Truth Used

Analytical Studies:
- Precision, Dilution Linearity, Spiked Recovery, Sensitivity, Hook Effect, Interference: Ground truth was based on known concentrations of PSA, characterized reference materials, or clinically established levels of interfering substances. For example, for dilution linearity, the "Expected Value" was the ground truth based on known dilutions. For spiked recovery, the "Concentration Added" was the ground truth.
Clinical Studies:
- Disease Cohorts (Normal, Benign, Malignant): Ground truth was based on patient diagnoses (e.g., "Benign prostate disease," "Prostate Cancer") established through clinical assessment (including other diagnostic tests and potentially pathology results, though not explicitly detailed for this specific aspect).
- Serial Monitoring: Ground truth for "Progression" or "No Progression" was based on "the clinical status of the patients as measured by other laboratory tests, patient interviews, physical examinations, and imaging studies of a variety of types." This is essentially patient outcomes data/clinical diagnosis as determined by a physician.

8. Sample Size for the Training Set

The document does not explicitly describe a separate "training set" for an algorithm in the way that would typically be detailed for an AI/ML device. This device is an immunoassay, and its performance characteristics (like linearity, precision, etc.) are established through traditional analytical and clinical validation studies, rather than machine learning model training on a distinct dataset. The "development" and "optimization" of the assay would typically involve internal samples and experiments, but these are not referred to as a "training set" in the context of this 510(k) submission.

9. How the Ground Truth for the Training Set Was Established

Since a "training set" for an AI/ML algorithm is not described, the method for establishing its ground truth is not applicable/provided. The analytical and clinical performance data presented here are for the validation and verification of the device's performance against established metrology principles and clinical outcomes.

Summary

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510(k) Summary

510(k) Number:

FREND™ PSA Plus on the FREND™ system K124056:

Summary Preparation Date:

December 28, 2012

Submitted by:

NanoEnTek, Inc. 12F, Ace High-end Tower, 235-2, Guro3-dong, Guro-gu Seoul, 152-740, Korea

Jimmy Chen

Owner/Operator Jimmy Chen (jim@nanoentek.com)

Contact:

Judith E Loebel Director, Clinical and Regulatory Affairs DOCRO, Inc. (CRO assigned as contact with the FDA)

Proprietary Names:

For the assay: FREND™ PSA Plus

For the instrument: FREND™ system

Common Names:

For the assay: Quantitative PSA immunoassay

For the instrument: Bench-top fluorometer with microprocessor

Regulatory Information:

Regulation section:

866.6010 Tumor Associated Antigen Immunologic Test System 862.2560 Fluorometer

Classification:

Class II

Panel:

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Immunology

Product Code(s):

LTJ Tumor Associated Antigen Immunologic Test System For the assay: For the instrument:

Predicate Device: TOSOH ST AIA-PACK PA (P910065/S004)

Intended Use:

The FREND™ PSA Plus performed on the FREND™ system, is a quantitative in vitro diagnostic test which measures total Prostate Specific Antigen (PSA) in human serum and plasma. The NanoEnTek FREND™ PSA Plus is designed for in vitro DIAGNOSTIC USE ONLY for the quantitative measurement of total Prostate Specific Antigen (PSA) in human serum, heparinized plasma, and EDTA plasma using the FREND™ System. This device is indicated for the serial measurement of total PSA in serum, heparinized plasma and EDTA plasma to be used as an aid in the management of patients with prostate cancer.

The FREND™ PSA Plus is indicated for use in clinical laboratories upon prescription by the attending physician as an aid to clinicians in managing patients with prostate cancer.

Technological Characteristics:

Material Provided - FREND™ PSA Plus - Catalog Number FRPS 025

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25 FREND™ PSA Plus cartridges The Test Cartridge is a disposable plastic device that . houses the reagents and contains an opening 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 sample is dropped into the specimen inlet located on the top left corner of the Test Cartridge. Sample and reagents interact before being analyzed by the FREND System fluorescence reader. The shelf-life of FREND PSA Plus cartridges is 12 months. One Cartridge contains:
Monoclonal anti-PSA1 48 ± 9.6 ng

Monoclonal anti-PSA2 144 ± 28.8 ng

Fluorescent particle 2.4 ± 0.48 µg

30 Disposable pipette tips (micro-pipettor optional) .
1 FREND™ PSA Plus Code Chip The OC Code Chip contains data to ensure the . performance of the FREND System's power, optical, and software systems when the QC Cartridge is inserted into the System. The QC Code Chip is inserted into the code chip port at the rear of the System. Each QC Code Chip is specific to its accompanying QC Cartridge. Each time a new lot is used, the PSA Code Chip that corresponds to that new lot must be inserted into the instrument and the previous PSA Code Chip must be removed. If the PSA Code Chip and the PSA Cartridge are not from the same lot, an error message will appear.
1 FREND™ PSA Plus Package Insert .
1 Product Certificate .
QC Case Storage box for the QC Cartridge and QC Code Chip .
Optional FREND™ System Pipettor device sheathed in a disposable single use plastic tip . used to transfer samples to the Cartridge.
Adaptor & Power cable used to supply power to the System .
FREND™ System User Manual .
FRENDIM System Quick Manual .
. USB drive 1.1
Optional printer Results of the test can be printed out using the optional printer. . Otherwise, they are displayed on the screen.

Commercially available controls from a variety of manufacturers are available that contain total PSA as a measured analyte. These controls are not provided with the assay cartridge.

The FREND™ System is not provided with the kit but is required for utilization of the FREND™ PSA Plus assay cartridge.

FREND™ System is a bench top fluorescence reader containing a touchscreen user interface. The System has a slot that accepts the FREND™ PSA Plus 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 through the RS232C interface.

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Performance Data Summary - Analytical Testing

ﺗ

To determine the analytical validity of the FREND™ PSA Plus, a series of studies have been performed as described below.

1. Estimates of imprecision: intra-assay, inter-assay and complex imprecision analyses were generated at three sites including the NanoEnTek, Inc. facility as well as at CLIA licensed facilities in the US. Imprecision was found to be acceptable at all three sites on all three lot numbers. The magnitude of the imprecision is not significantly different than that seen with other devices of this type.
  Precision data was determined as described in the CLSI protocol EP5-A2. Three clinical samples were assayed in replicates of two at two separated times per day for twenty days using a single lot cartridge. Results are shown below in table format. All elements of this testing process met the specifications set. Repeatability and Within-laboratory precision on samples with measured concentrations from 0.1 to 1.0 ng/mL was less than 15%. At concentrations >1.0 ng/mL but < 25 ng/mL, Repeatability and Within-laboratory precision was less than 10%.

Sample	Mean PSA	Repeatability		Between-run		Between-day		Within-laboratory
	(ng/mL)	SD	CV(%)	SD	CV(%)	SD	CV(%)	SD	CV(%)
1	0.186	0.026	14.20	0.005	2.9	0.004	2.3	0.027	14.6
2	2.757	0.221	8.0	0.132	4.8	0.071	2.6	0.268	9.7
3	16.625	1.407	8.5	0.000	0.0	0.449	2.7	1.477	8.9

Clinical Sample Precision

Three different lots of FREND™ PSA Plus were evaluated at three geographically diverse sites. Four replicates each of Material A, Material B, and Material C and two replicates of QC 1, QC 2 and QC 3 were evaluated in each of two runs performed for five days at each site. A total of 40 results on each material were generated at each of the three sites yielding a grand total of 120 replicates of each material. The data was analyzed using a CLSI format from EP5-A2 for an ANOVA analysis. Instrument-to-Instrument is the same as Site-to-Site in this scenario. As can be seen from the table below, the largest source of variation is the cartridge which would be the expected result. The FREND™ PSA Plus cartridge is a single use cartridge that contains all the reagents within the cartridge necessary to support the reactions.

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%CV by Material for Multi-Site, Multi-Lot Imprecision
Material
Variation Source	MAT A(0.29 ng/mL)	MAT B(3.67 ng/mL)	MAT C(18.33 ng/mL)	QC 1(0.30 ng/mL)	QC 2(2.93 ng/mL)	QC 3(20.25 ng/mL)
Site-to-Site	3.50%	1.57%	1.67%	3.47%	1.61%	2.06%
Day-to-Day	0.00%	0.99%	1.21%	0.00%	0.00%	0.00%
Lot-to-Lot	9.12%	3.16%	7.01%	6.08%	4.30%	6.00%
Inter- cartridge	18.45%	6.81%	7.94%	20.03%	6.17%	7.49%
Total	20.87%	7.74%	10.79%	21.22%	7.69%	9.81%

2. Dilution Linearity & Recovery:

To demonstrate the linearity of the assay, a serum sample pool with an elevated total PSA (34 ng/mL) was diluted to a total of seven levels according to the dilution protocol outlined in CLSI-EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach. In addition, a "neat" sample (no dilution) and a "zero" sample were also be run. At each dilution level, the samples were tested in 6 replicates to determine the observed value of PSA. Percent recovery was calculated by comparing the observed PSA result with the expected value. Recoveries within + 10% of the expected results for the overall mean recovery at concentrations above 1.0 ng/mL for each sample were considered acceptable proof of this performance goal.

Image /page/4/Figure/3 description: This image is a scatter plot with a linear trendline. The x-axis is labeled "Expected value (ng/ml)" and ranges from 0 to 40. The y-axis is labeled "FREND PSA Plus (ng/ml)" and ranges from 0 to 40. The equation of the trendline is y = 0.9856x + 0.1324, and the R-squared value is 0.9992.

Dilution Linearity FREND™ PSA Plus

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No.	Dilution	TEST 1	TEST 2	TEST 3	TEST 4	TEST 5	TEST 6	MEAN(ng/mL)	SD	CV%	ExpectedValue	%Recovery
Blank	0.000	0.00	0.00	0.10	0.00	0.00	0.10	0.033			0.0
1	0.125	4.63	4.65	4.08	4.27	4.59	4.26	4.413	0.241	5.5	4.3	103.8
2	0.250	8.85	8.65	9.24	8.18	8.67	7.74	8.555	0.526	6.1	8.0	106.9
3	0.375	13.30	12.33	12.71	13.91	12.61	11.58	12.740	0.802	6.3	12.8	99.9
4	0.500	18.34	16.89	18.22	16.07	17.56	18.57	17.608	0.972	5.5	17.0	103.6
5	0.625	21.44	22.49	19.39	19.32	20.67	22.78	21.015	1.490	7.1	21.3	98.9
6	0.750	23.78	25.42	27.13	26.88	24.91	25.93	25.675	1.255	4.9	25.5	100.7
7	0.875	28.04	34.33	27.12	29.51	27.36	26.38	28.790	2.913	10.1	29.8	96.8
High	1.000	39.98	35.22	31.74	27.43	28.32	40.09	33.797	5.561	16.5	34.0	99.4

1. Spiked Recovery: a serum pool from females (initial tPSA concentration < 0.01 ng/mL) was spiked with three (3) different known levels of PSA across the range of the assay (0.1 to 25 ng/mL). After spiking, samples were assayed during the same run before and after spiking. The percentage of tPSA recovered was compared to the theoretical amount spiked into the samples. Recoveries within 10% of the expected result for the overall mean recovery for a given sample were considered acceptable proof of this performance goal.

ConcentrationAdded (ng/mL)	ObservedConcentration(ng/mL)	Recovery(%)
	1.06	98.3
1.08	1.09	100.7
1.08	1.04	96.7
4.34	4.42	101.8
4.34	4.35	100.3
4.34	4.27	98.4
12.81	13.58	106
12.81	12.04	94

Spiked Recovery FREND™ PSA Plus

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	11.82	92.3
	24.26	95
25.53	24.67	96.6
	26.90	105.4

The percentage recovery ranged from 92.3% to 105.4%. The results are within the acceptance region.

1. Analytical Sensitivity (Limits of Detection or LOD/LOQ): Determination of the limit of detection/limit of quantitation was performed according to procedures outlined in CLSI-EP17-A: Protocols for Determination of Limits of Detection and Limits of Quantitation. In the determination, 5 samples without PSA (blank samples) and 5 samples with a low PSA concentration were tested in the assay in 12 replicates for each sample. The 95th percentile value for the blank samples was 0.04 ng/mL. The 5th percentile value for the low PSA samples was 0.055 ng/mL. At a PSA value of 0.1 ng/mL, there were no blank samples (without PSA) labeled as detected by the assay and 50% of low PSA sample values (LOD samples) had a PSA value below 0.1 ng/mL. The LOD/LOQ limit as determined by this testing is 0.1 ng/mL PSA.
1. High Dose Hook Effect Testing (Prozone Detection): The presence of high dose hook effect was tested by analyzing a concentrated sample of purified PSA antigen both neat and on dilution. No High Dose Hook effect was seen in samples with a PSA concentration as high as 1200 ng/mL.
1. Interfering Substances and Assay Specificity: Prostatic acid phosphatase and kallikrein (not otherwise described) were evaluated for potential cross-reactivity with the FREND PSA Plus at 10 ng/mL and 15 ng/mL respectively utilizing the instructions recommended by CLSI protocol EP7-A. No significant cross-reactivity was found.

Interference studies on endogenous serum substances were performed using the FREND PSA Plus according to the recommendations in the CLSI protocol EP7-A. Interference is defined to be recovery outside of 15% of the known specimen mean concentration. Lack of interference (recovery from 85% to 115% of the expected) is considered acceptable performance. The following describes the interferents tested and the maximum concentration without interference at the total PSA concentrations evaluated:

· Added hemoglobin (up to 500 mg/dL) does not interfere with the assay. Average recovery when added to serum containing tPSA at 1.0 and 4.0 ng/mL was 97.25%.
· Added conjugated bilirubin (up to 20 mg/dL) does not interfere with the assay. Average recovery when added to serum containing tPSA at 1.0 and 4.0 ng/mL was 98.2%.

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Added gamma globulin (Total Protein) up to 5.0 g/dL does not interfere with the . assay. Average recovery when added to serum containing tPSA at 1.0 and 4.0 ng/mL was 106.3%.
· Added triglyceride up to 3 grams/dL does not interfere with this assay. Average recover when added to serum containing tPSA at 1.0 and 4.0 ng/mL was 101.5%.

No interference was found from endogenous materials such as bilirubin, triglycerides, hemoglobin, or added protein at a level above what is found in the usual clinical laboratory specimen.

Drugs commonly given to patients with prostate cancer and some common over-thecounter medications were also tested using CLSI protocol EP7-A.

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Substance	InterferentConcentration tested	Average %Recovery
flutamide	10 µg/mL	94.5%
Diethylstilbestrol(DES)	5 µg/mL	103.8%
goserelin	40 ng/mL	103.2%
tamsulosin	100 ng/mL	98.85%
acetaminophen	250 ng/mL	100.45%
acetylsalicylicacid	600 µg/mL	95.85%
leuprolide	275 ng/mL	101.5%
ibuprofen	500 µg/mL	102.25%
finasteride	250 ng/mL	93.6%
docetaxel	10 µg/mL	114.45%

Drug Interference Testing FREND™ PSA Plus

No interference outside the acceptable recovery range (100 ± 15%) was found.

1. Method comparison: Comparison studies were done in the DOCRO, Inc. laboratory testing facility, Orion Laboratory, Inc., at the time the clinical samples enrolled for this study were evaluated. The predicate device and the FREND™ system were run at the same time, side-by-side on the bench so samples were tested simultaneously using the same aliquot of sample. Regression analysis was performed on the data pairs obtained from prostate cancer samples taken at a single point (n = 85) and the earliest sample from 75 subjects undergoing serial monitoring for prostate cancer. Any samples reading above either of the linearity limits of the predicate or proposed assay was diluted and the final result determined by multiplying the dilution factor by the answer obtained on that dilution. Statistics were generated on the samples from prostate cancer subjects (n=160). For those results whose tPSA was less than 25 ng/mL, the linearity limit of the FREND™ PSA Plus assay, comparison was performed using Passing-Bablok regression analysis.

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Image /page/9/Figure/0 description: The image shows a scatter plot with a regression line. The x-axis is labeled as 'Predicate Device (ng/mL)' and ranges from 0 to 30. The y-axis ranges from 0 to 30. The regression line equation is 'y=0.94x-0.006'.

The two methods compared well within the specifications usually accepted in the clinical laboratory.

Sample Matrix Comparison Study: The FREND™ PSA Plus designates multiple 8. matrices as being acceptable sample types. Lithium heparin and EDTA plasma are specified in addition to serum as appropriate sample types. Matrix comparison studies were performed in which the various matrices above plus sodium citrate were compared with respect to the PSA result in a serum matrix. Deming regression and Passing-Bablok were performed. Concentrations of PSA across the measuring range were used. 36 matched sets were collected, aliquoted, frozen and then run once all collections were completed.
To determine the statistical relationship between the PSA concentrations of the plasma specimens and that of the serum specimen, linear regression and Passing-Bablok regression analyses were performed. Equivalence of repeatability in duplicate runs for all matrices and equivalence of variance between serum and each of three different plasma matrices in three partitions were also evaluated. An assessment of the repeatability of variance for each matrix type was performed to ensure that variation in serum samples was equivalent to variation in each plasma type. Analysis showed that EDTA heparin, lithium heparin and serum are all acceptable matrices. Sodium citrate showed unacceptable performance.
1. Reagent Stability Studies: Real time stability studies of the FREND PSA Plus Test Cartridge were conducted over 15 months on three different lots. The Test Cartridges were either stored at 2 - 8°C or at room temperature. At each of three month intervals, the Test Cartridge was assayed with five standard specimens. The results using both the refrigerated and the room temperature stored cartridges for the five standard specimens yielded results with a %CV of less than 10% meeting the acceptance criteria. Reagent

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studies showed that the cartridges for FREND™ PSA Plus are good for at least one year from date of manufacturer if stored appropriately as directed.

10. Performance Data Summary - Clinical Testing

A total of 1219 evaluable clinical serum samples were assessed during this study. Results were obtained on both the FREND™ PSA Plus and the predicate device. Sample distributions across various cohorts of samples: Normal, Benign and Malignant, were studied. Testing of ambulatory male subjects fifty years old and older who reported themselves as healthy without any known illnesses, diseases or conditions was performed. The cohorts shown in the following table were assembled to determine the distribution of values in benign diseases that may be co-existent in patients with confirmed prostate cancer. Prospectively collected stored samples were utilized for this study. .

Cohort	Number
Benign prostate disease (BPH, prostatitis, etc.)	104
Benign Diseases of the GI tract	107
Subjects with Diabetes of any type	97
Cardiovascular Disease/Hypertension	102
Total	410

The following patient cohorts were assembled to determine the distribution of PSA values in patients with known malignancies (a mixture of treated and untreated are represented).

Cohort	Number
Lung/Liver Cancer	52
Gall Bladder/Gastric/PancreaticCancer	31
Prostate Cancer (Divided by GleasonScore)	85
Colorectal Cancer	89
Other Cancers	45
Total	302

Normal male cohort: For the FREND PSA Plus assay in normal men age 50 years or older the following graph shows the distribution of PSA in these normal men.

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Image /page/11/Figure/0 description: This image is a histogram titled "Histogram with Reference Interval". The y-axis is labeled "Frequency" and ranges from 0 to 50. The x-axis is labeled "FREND PSA Plus Results (ng/mL)" and ranges from 0 to 4. The histogram shows the frequency of different PSA levels, with the highest frequency between 1 and 2 ng/mL.

The 95th percentile value using the FREND PSA Plus assay is well below a value of 4.0 ng/mL. The upper 95th percentile value for the same cohort of men using the predicate assay was similar.

and Various Malignant Disease States
	N	0 - 4.0ng/mL	4.1 - 10.0ng/mL	10.1 - 20ng/mL	20.1 - 40ng/mL	>40ng/mL
Healthy Subjects	196
Men ≥ 50 yrs.	196	100%	0%	0%	0%	0%
Benign Disease/Cond*	410
Benign Prostate	104	56.73%	25.96%	11.54%	3.85%	1.92%
Diabetes	97	95.88%	3.09%	1.03%	0.00%	0.00%
HTN/Heart Disease	102	95.10%	4.90%	0.00%	0.00%	0.00%
Benign GI	107	94.4%	4.67%	0.00%	0.93%	0.00%

Malignant Diseases*	302
Prostate Cancer**	85	40.00%	38.82%	12.95%	2.35%	5.88%
Gleason Score 5-6	43	51.16%	44.19%	2.38%	2.38%	0%
Gleason Score 7	31	35.48%	38.72%	19.35%	0%	6.45%
Gleason Score 8-9	11	9.09%	18.18%	36.36%	9.09%	27.27%
Lung/Liver Cancer	52	98.08%	0%	1.92%	0%	0%
GB,Gastric,Pancreatic	31	100%	0%	0%	0%	0%
Colorectal Cancer	89	94.38%	4.49%	1.13%	0%	0%
Other Cancers	45	97.78%	2.22%	0%	0%	0%

Distribution of Serum FRENDTM PSA Plus Concentrations in Healthy, Benign
and Various Malignant Disease States

*Treated and untreated subjects

TOTAL Subjects

** Serial samples are not included in this cohort.

Performance between the two assay systems did not show any clinically significant differences.

Serially monitored subjects: As an important part of the clinical studies performed to characterize the FREND™ PSA Plus, serial samples collected longitudinally from patients previously diagnosed with prostate cancer and treated in a variety of ways over the clinical

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course of their disease (including prostatectomy, radioactive seeds, external beam radiation, chemotherapy, hormone therapy alone or in combination) were assayed for tPSA with the FREND™ PSA Plus on the FREND™ system. The same samples were also measured for tSA by the predicate PSA method.

For each point to point in a sample serial set, the change in the tPSA concentration was compared to the change in the clinical status of the patients as measured by other laboratory tests, patient interviews, physical examinations, and imaging studies of a variety of types.

These changes in the tPSA marker concentration were defined as significant or not by multiplying the overall %CV of the assay at the midrange (as determined by the test imprecision study) by a factor of 2.5 to define a percentage change difference higher than would be expected because of assay imprecision. For the FREND™ PSA Plus assay with an overall mid-range %CV of 8%, significance was set at a change in excess of 20%. Any increase in value from one time period to the next that did not exceed 20% was logged as ≤ 20% Change. For the predicate method, significant percentage change was set at a change > 8.5%. This was calculated using that method's published overall mid-range CV of 3.4% x 2.5.

Disease status for the patients was determined by the physician. This Disease Status was used to determine Progression or No Progression from a clinical perspective. The first table below shows the progressions and non-progressions as determined for the FREND™ PSA Plus results for all seventy five subjects compared to the Clinical Status changes. The second table shows the same comparison for the other FDA cleared assay. There are a total of 236 such determinations for each assay.

Point to Point for FRENDTM PSA Plus
% Change in PSA Value	Progression	No Progression	Total
Change≥ 20.0%	84	46	130
Change< 20%	25	81	106
Total	109	127	236

Point to Point for Another FDA Cleared PSA Assay
% Change in PSA Value	Progression	No Progression	Total
Change≥ 8.5%	88	45	133
Change< 8.5%	21	82	103
Total	109	127	236

Below is a chart comparing the concordances of the FREND™ PSA Plus assay and the TOSOH ST AIA-PACK PSA.

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Concordance	FREND	95% CI*	FDA Cleared PSA	95% CI*
Positive	77.06%	69.02% to84.85%	80.73%	73.19% to88.03%
Negative	63.78%	55.30% to71.97%	64.57%	56.25% to75.59%
Total	69.92%	63.98% to75.85%	72.03%	66.10% to77.54%

Concordance FREND™ PSA Plus and TOSOH ST AIA-PACK™ PSA

Confidence Intervals are based on 10,000 resamples of the patient data

Based on the 95% confidence intervals, there are no differences between the concordances for the FREND™ PSA Plus assay and TOSOH ST AIA-PACK™ PSA assay. Positive, negative and overall concordances determined on the serial sets of samples for the two methods showed no significant difference in the ability of the assay to mirror the patient clinical status.

Substantial Equivalence

The FREND™ PSA Plus is as safe and effective as the "Predicate Device", the TOSOH ST AIA-PACK PA assay. FREND™ PSA Plus has a similar Intended Use and Indications for Use for monitoring of prostate cancer patients, similar technological and performance characteristics, and principles of operation as its predicate device. The differences between the FREND™ PSA Plus and its predicate device raise no new. issues of safety or effectiveness. Performance data, analytical and clinical, demonstrate that the FREND™ PSA Plus is as safe and effective as the "Predicate Device".

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/14/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol resembling three stylized waves or ribbons, arranged in a descending order from left to right.

Public Health Service

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

May 29, 2013

NANOENTEK INC. C/O JUDITH E. LOEBEL DIRECTOR, CLINICAL AND REGULATORY AFFAIRS DOCRO INC. 1 JACKS HILL ROAD SUITE A & B OXFORD CT 06478

Re: K124056

Trade/Device Name: FREND™ PSA Plus on the FREND™ System Regulation Number: 21 CFR 866.6010 Regulation Name: Tumor-associated antigen immunological test system Regulatory Class: II Product Code: LTJ, KHO Dated: May 18, 2013 Received: May 24, 2013

Dear Ms. Loebel:

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. Iisting 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 (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance 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. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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/Safetv/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 Small Manufacturers, International and Consumer Assistance 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.

Sincerely vours,

Maria Mi Mhan -S

Maria M. Chan, Ph.D.

Director

Division of Immunology and Hematology 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): K124056

Device Name: FREND™ PSA Plus on the FREND™ system

Indications For Use:

The FREND™ PSA Plus is indicated for use in clinical laboratories upon prescription by the attending physician as an aid to clinicians in managing patients with prostate cancer.

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH; Office of In Vitro Diagnostics and Radiological Health (OIR)

Maria M. @han -S

Division Sign-Off Office of In Vitro Diagnostics and Radiological Health

510(k):

Regulation Number and Section

§ 866.6010 Tumor-associated antigen immunological test system.

(a)
Identification. A tumor-associated antigen immunological test system is a device that consists of reagents used to qualitatively or quantitatively measure, by immunochemical techniques, tumor-associated antigens in serum, plasma, urine, or other body fluids. This device is intended as an aid in monitoring patients for disease progress or response to therapy or for the detection of recurrent or residual disease.(b)
Classification. Class II (special controls). Tumor markers must comply with the following special controls: (1) A guidance document entitled “Guidance Document for the Submission of Tumor Associated Antigen Premarket Notifications (510(k)s) to FDA,” and (2) voluntary assay performance standards issued by the National Committee on Clinical Laboratory Standards.