For the quantitative measurement of human chorionic gonadotropin (hCG) and its ß-subunit in human serum and plasma (heparin and EDTA) using the VITROS 5600 Integrated System to aid in the early detection of pregnancy.

Device Description

The VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack (test) is performed using the VITROS Immunodiagnostic Products Total β-hCG II Reagent Pack and VITROS Immunodiagnostic Products Total B-hCG II Calibrators on the VITROS 5600 Integrated System.

An immunometric immunoassay technique is used, which involves the reaction of human chorionic gonadotropin (hCG) present in the sample with a microwell coated with biotinylated Antibody (mouse monoclonal anti-ß-hCG) bound to streptavidin, and a Horseradish Peroxidase (HRP)-labelled antibody conjugate (mouse monoclonal anti-ß-hCG). Unbound materials are removed by washing.

The bound HRP conjugate is measured by a luminescent reaction. A reagent containing luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent, is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent (a substituted acetanilide) increases the level of light produced and prolongs its emission. The light signals are read by the system. The amount of HRP conjugate bound is directly proportional to the concentration of hCG present in the sample.

VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack contains:
1 reagent pack containing:

100 coated wells (antibody, mouse monoclonal anti-ß-hCG, binds >600 mIU hCG/well)
14.4 mL assay reagent (buffer containing mouse serum, bovine serum albumin, bovine gamma globulin and . antimicrobial agent)
19.2 mL conjugate reagent (HRP-mouse monoclonal anti-β-hCG, binds ≥4005 mIU hCG/mL) in buffer with ● bovine serum albumin and antimicrobial agent.

VITROS Total ß-hCG II Calibrators contains:

3 sets of VITROS Total ß-hCG II Calibrators 1, 2 and 3, (freeze-dried, recombinant hCG in human plasma with antimicrobial agent, reconstitution volume 1.0 mL), nominal values 0; 3,000; 14,000 mIUmL (U/L)
24 calibrator bar code labels (8 for each calibrator)

AI/ML Overview

Here's an analysis of the acceptance criteria and study information based on the provided text, structured according to your request:

1. A table of acceptance criteria and the reported device performance

The document does not explicitly present a dedicated "acceptance criteria" table for all non-clinical tests. Instead, it describes each test (e.g., Precision, Detection Capability, Linearity) and then presents the results, implying that the results met internal acceptance criteria for substantial equivalence to the predicate device. Therefore, I will derive the "acceptance criteria" from the stated goals or industry guidelines mentioned for each test and then present the reported performance.

Test Category	Acceptance Criteria (Inferred/Stated)	Reported Device Performance
Stability (Shelf-life)	Consistency with methods based on CLSI EP25-A, supporting the claimed shelf-life.	52-week shelf-life supported by four runs on each of 3 Lots at monthly intervals.
Stability (On-board)	Consistency with methods based on CLSI EP25-A, supporting the claimed on-board stability.	Supported by evaluation of three Lots stored opened refrigerated for up to 12 weeks, with four runs on each Lot at each time-point for fresh and open samples. All results acceptable and support 6 weeks on-board stability.
Precision (Repeatability)	Evaluation consistent with CLSI document EP05-A3. (Specific %CV or SD targets not explicitly stated in this section, but implied by acceptable performance.)	Repeatability (SD/%CV): - 5.33 mIU/mL: 0.095 (1.80%)- 15.46 mIU/mL: 0.181 (1.20%)- 45.67 mIU/mL: 0.666 (1.50%)- 297.34 mIU/mL: 4.970 (1.70%)- 4708.50 mIU/mL: 82.29 (1.70%)- 9651.00 mIU/mL: 286.71 (3.00%)
Precision (Within Lab)	Evaluation consistent with CLSI document EP05-A3. (Specific %CV or SD targets not explicitly stated in this section, but implied by acceptable performance.)	Within Lab (SD/%CV): - 5.33 mIU/mL: 0.281 (5.30%)- 15.46 mIU/mL: 0.706 (4.60%)- 45.67 mIU/mL: 1.560 (3.40%)- 297.34 mIU/mL: 8.640 (2.90%)- 4708.50 mIU/mL: 151.22 (3.20%)- 9651.00 mIU/mL: 464.48 (4.80%) Further breakdown including Between Lot shows Total %CV up to 6.4%
Detection Capability (LoB)	Evaluation consistent with CLSI document EP17-A2. Supporting the claimed LoB of 0.05 mIU/mL.	Representative LoB is 0.00 mIU/mL (IU/L), which supports the claimed LOB of 0.05 mIU/mL.
Detection Capability (LoD)	Evaluation consistent with CLSI document EP17-A2. Supporting the claimed LoD of 0.70 mIU/mL.	Representative Limit of Detection (LoD) is 0.21 mIU/mL (IU/L), which supports the claimed LoD of 0.70 mIU/mL (IU/L).
Detection Capability (LoQ)	Designed to be less than or equal to the claimed low end of the measuring range (2.39 mIU/mL) at 20% CV. Consistent with CLSI document EP17 (Total Error approach).	Representative LoQ using the Total Error approach was 2.32 mIU/mL (IU/L). Claimed LoQ verified at 2.39 mIU/mL (IU/L).
Linearity	Established in accordance with CLSI guideline EP06 2nd edition. Results support linearity across the specified range.	Supported linearity from 1.51 mIU/mL (IU/L) to 15695 mIU/mL (IU/L). For Lot 2643, Slope was 0.991 (38.6 to 15695) and 1.025 (1.51 to 51.18); R2 was 0.996 and 0.999 respectively; % Recovery ranged from 92.2% to 112.7%.
Measuring Range	The device is expected to have a measuring range on the VITROS 5600 system.	2.39*-15,000 mIU/mL (IU/L).
Matrix Comparison	Serum and plasma (Lithium-Heparin and K2-EDTA) specimen matrices determined to be equivalent. Results met acceptance criteria for comparison between serum and plasma spanning the expected measuring interval.	Weighted Deming Regression: Li-Hep Plasma (Slope 0.978, Corr. Coef r 0.999), K2-EDTA Plasma (Slope 0.978, Corr. Coef r 0.999). Serum and plasma (Li-Heparin and K2-EDTA) found suitable matrices.
Analytical Specificity (Known Interferents)	Evaluated following CLSI EP07 and EP37. None found to cause a bias of >10% at specified hCG concentrations.	Over 30 common substances (e.g., Acetaminophen, Bilirubin, Biotin, Hemoglobin, Triglycerides) tested at high concentrations (e.g., Acetaminophen 15.6 mg/dL, Hemoglobin 1000 mg/dL, Triglycerides 1500 mg/dL) showed no bias >10% at hCG concentrations of 5.00, 50.00, and 10,000 mIU/mL.
Analytical Specificity (Cross-Reactivity)	Evaluation for cross-reactivity with FSH, LH, and TSH in hCG negative samples and samples with ~25 mIU/mL hCG.	In hCG negative samples, FSH, LH, and TSH (at 400 mIU/mL and 200 mIU/mL respectively) were Not Detectable (ND). With a pool at ~25 mIU/mL hCG, % Cross Reactivity was: FSH (0.6%), LH (-0.3%), TSH (0.4%).
Expected Values (Adult Reference Interval)	Validated following CLSI document EP28-A3c. Distribution of hCG values from normal healthy non-pregnant blood donors shows equivalency to the predicate device's expected values claim.	Original Claim: Total 290 samples, Mean 0.56, Min 0.00, Max 6.66, 2.5th Percentile 0.00, 97.5th Percentile 4.83. Updated Pack: Total 180 samples, Mean 0.47, Min 0.00, Max 5.61, 2.5th Percentile 0.00, 97.5th Percentile 3.73. Demonstrated equivalency.
Method Comparison (Accuracy)	Evaluated consistent with CLSI guideline EP09c. Comparison with predicate device (VITROS Immunodiagnostic Products Total β-hCG II reagent pack, K063720).	n=135 samples. Slope 0.99 (95% CI: 0.977 to 0.995). Correlation Coefficient 0.998. Intercept -0.0215 (95% CI: -0.160 to 0.117). (Comparing VITROS Total ß-hCG II (GEM.1076A) against predicate VITROS Total B-hCG (GEM.1076)). Consistent with graphs indicating strong agreement.

2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)

Stability Studies:
- Long-term: 3 Lots evaluated.
- On-board: 3 Lots evaluated.
Precision:
- Single Lot Precision: 6 precision fluids. 2 replicates per run, 2 runs per day for 20 days. Total of 80 data points per fluid.
- Additional Precision Analysis Summary: 6 samples (PP1-6).
- Multiple System (Reproducibility) Study: 6 precision fluids (RP1-6). 5 replicates per run, 1 run per day for 5 days. Tested on 3 VITROS 5600 integrated systems.
Detection Capability (LoB): 4 serums containing no measurable hCG. Study design: 2 replicates per run, 2 runs per day over 5 test days (20 reps per test fluid x 4 fluids = 80 replicates) x 3 lots = 240 total replicates.
Detection Capability (LoD & LoQ): 5 LoD samples targeted at 1 to 5 times LoB. 4 LoQ fluids for Total Error method. All samples run using 3 reagent lots on one VITROS 5600 System. 6 replicates per run, 2 runs per day over 5 test days (60 reps per fluid x 5 fluids = 300 replicates) x 3 lots = 900 total replicates.
Linearity: Two series: one across entire range (pools 1a to 10a) and one in lower range (pools 1b to 10b). 10 replicates of pools 1a/10 and 10a/10b, and 5 replicates of pools 2a/2b to 9a/9b. Run on one VITROS 5600 Integrated System.
Matrix Comparison: 41 samples each for Lithium-Heparin Plasma and K2-EDTA Plasma.
Analytical Specificity (Known Interferents): Not specifically quantified, but refers to "compounds tested" at three hCG concentrations.
Analytical Specificity (Cross-Reactivity): hCG negative samples and hCG pools at approximately 25 mIU/mL were used for spiking test substances (FSH, LH, TSH).
Expected Values (Adult Reference Interval):
- Original Claim (from predicate): 290 total samples (98 normal male, 123 normal female, 69 post-menopausal).
- Updated Pack Validation: 180 normal healthy non-pregnant blood donors (60 Normal Male, 60 Normal Female, 60 Post-Menopausal).
Method Comparison: 135 human serum samples.

Data Provenance:

The document states that human serum samples for method comparison were "obtained from certified vendors."
Precision fluids mentioned as "pooling female serum samples."
"Normal healthy non-pregnant blood donors" for expected values.
"Neat serum samples containing low levels of endogenous hCG" for LoD samples.
The overall context is non-clinical laboratory testing. No specific country of origin or retrospective/prospective status is explicitly stated for the patient samples used, though the tests themselves were conducted in a laboratory setting per CLSI guidelines. The stability studies and many precision studies seem to be prospective evaluations of the manufactured product. Patient samples, however, are often retrospectively collected from vendors in such studies unless specified.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)

This device is an in-vitro diagnostic (IVD) test for quantitative measurement of hCG. The "ground truth" for such devices is established through reference methods, traceability to international standards, and comparison with legally marketed predicate devices, rather than expert human interpretation of images or clinical outcomes.

Traceability: The device's calibration is traceable to "in-house reference calibrators, which have been value-assigned with reference to the 4th International Standard (NIBSC 75/589)." This international standard serves as the "ground truth" for the quantitative measurement.
Method Comparison: The predicate device (VITROS Immunodiagnostic Products Total β-hCG II, K063720) serves as the comparative "ground truth" for assessing equivalence of performance with patient samples.

Therefore, there were no human "experts" (like radiologists) establishing ground truth in the way described for image-based diagnostic AI. The "ground truth" is defined by established metrological standards and comparative testing against a cleared predicate.

4. Adjudication method (e.g. 2+1, 3+1, none) for the test set

Not applicable. This is an IVD device for quantitative measurement based on chemical reactions and instrumental readings. There is no human interpretation or subjective assessment of results that would require an adjudication method among experts.

5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

Not applicable. This is an IVD device, not an AI-assisted diagnostic tool that aids human readers in image interpretation or clinical decision-making. No MRMC study was conducted.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

This device itself is a "standalone" analytical system in the context of laboratory testing. The VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack, used on the VITROS 5600 Integrated System, performs the quantitative measurement of hCG without immediate human intervention in the assay process itself. The "performance" sections (Precision, Detection Capability, Linearity, etc.) describe its standalone analytical performance.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

The ground truth for this IVD device is primarily:

International Reference Standards: Specifically, the 4th International Standard (NIBSC 75/589) for hCG, to which the device's calibration is traceable.
Comparison to a Legally Marketed Predicate Device: The performance of the new device is compared to the predicate device (VITROS Immunodiagnostic Products Total β-hCG II Reagent Pack, K063720), which established its own accuracy and reliability.
CLSI Guidelines: Various CLSI (Clinical and Laboratory Standards Institute) documents (e.g., EP05-A3 for Precision, EP17-A2 for Detection Capability, EP06 for Linearity, EP07/EP37 for Interferents, EP28-A3c for Reference Intervals, EP09c for Method Comparison) define the acceptable methodologies for establishing truth and performance in laboratory diagnostics.

8. The sample size for the training set

This document describes a non-AI IVD device. There is no "training set" in the machine learning sense. The device is a chemical reagent pack used on an analyzer. Its "training" equivalent relies on the design, manufacturing tolerances, and calibration traceable to international standards.

9. How the ground truth for the training set was established

Not applicable, as there is no "training set" in the context of an AI/ML algorithm. The "ground truth" for the device's fundamental function (accurate hCG measurement) is established through its traceability to international reference materials and validation against the predicate device, as detailed in point 7.

Summary

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

Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

May 8, 2024

Ortho Clinical Diagnostics Rebecca Lewis Senior Regulatory Affairs Associate Felindre Meadows Pencoed Bridgend, CF35 5PZ United Kingdom

Re: K233581

Trade/Device Name: VITROS Immunodiagnostic Products Total B-hCG II Reagent Pack Regulation Number: 21 CFR 862.1155 Regulation Name: Human Chorionic Gonadotropin (HCG) Test System Regulatory Class: Class II Product Code: DHA Dated: March 26, 2024 Received: March 26, 2024

Dear Rebecca Lewis:

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 (the 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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

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

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Paula V. Caposino -S

Paula Caposino, Ph.D. Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

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

Indications for Use

510(k) Number (if known) K233581

Device Name

VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack

Indications for Use (Describe) For in vitro diagnostic use only.

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

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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

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

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

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

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

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

VITROS Immunodiagnostic Products Total β-hCG II Reagent Pack Traditional 510(k)

510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92. The assigned 510(k) number is: K233581.

Submitter's Information

Ortho Clinical Diagnostics

Felindre Meadows.

Pencoed,

UK CF35 5PZ Phone: +44 (0) 7971 427649

Fax: 585-453-4110 Contact Person: Rebecca Lewis

Preparation Date

May 08, 2024

Device Proprietary Name(s)

VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack

Common Name(s) VITROS Total ß-hCG II Reagent Pack

Classification Names

ProductCode	Class	Regulation Section	Panel
DHA	II	21 CFR 862.1155	Clinical Chemistry

Predicate Device(s)

Predicate Device	FDA 510(k) Number
VITROS Immunodiagnostic Products Total β-hCG IIReagent Pack	K063720

Device Description

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

VITROS Immunodiagnostic Products Total β-hCG II Reagent Pack Traditional 510(k)

VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack contains:

1 reagent pack containing:

100 coated wells (antibody, mouse monoclonal anti-ß-hCG, binds >600 mIU hCG/well)
14.4 mL assay reagent (buffer containing mouse serum, bovine serum albumin, bovine gamma globulin and . antimicrobial agent)
19.2 mL conjugate reagent (HRP-mouse monoclonal anti-β-hCG, binds ≥4005 mIU hCG/mL) in buffer with ● bovine serum albumin and antimicrobial agent.

VITROS Total ß-hCG II Calibrators contains:

3 sets of VITROS Total ß-hCG II Calibrators 1, 2 and 3, (freeze-dried, recombinant hCG in human plasma with antimicrobial agent, reconstitution volume 1.0 mL), nominal values 0; 3,000; 14,000 mIUmL (U/L)
24 calibrator bar code labels (8 for each calibrator)

Intended Use Statement(s):

For in vitro diagnostic use only.

Comparison to Predicate Devices

The following table provides a summary of the key features of the new device assessed against the predicate.

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

Pack Traditional 510(k)
DeviceCharacteristic	Predicate Device	Modified Device
Intended Use	For in vitro diagnostic use only.For the quantitative measurement ofhuman chorionic gonadotropin (hCG) andits β-subunit in human serum and plasma(heparin and EDTA) using the VITROSECi/ECiQ/3600 ImmunodiagnosticSystems and the VITROS 5600/XT 7600Integrated Systems to aid inthe early detection ofpregnancy.	For in vitro diagnostic use only.For the quantitative measurement of humanchorionic gonadotropin (hCG) and its β-subunit in human serum and plasma (heparinand EDTA) using the VITROS 5600Integrated System to aid in the early detectionof pregnancy.
Antibody	Mouse Monoclonal anti-β-hCG.	Same.
Sample Type	Serum and Plasma.	Same.
Sample Volume	40 μL.	Same.
Traceability	Traceable to in-house referencecalibrators, which have beenvalue assigned with reference tothe 4th International Standard(NIBSC 75/589).	Same.
MeasuringRange	Analytical Measuring Interval:$2.39-15,000$ mIU/mL (IU/L)	Same.
DetectionLimit	LOB: $0.05$ mIU/mL (IU/L)LOD: $0.70$ mIU/mL (IU/L)	LOB:SameLOD: Same.LOQ: $2.39$ mIU/mL (IU/L)
Calibrator Levels	3.	Same.
DeviceCharacteristic	VITROS Immunodiagnostic ProductsTotal β-hCG II reagent pack, K063720,cleared 09 April 2007	VITROS Immunodiagnostic Products Total β-hCG IIreagent pack

VITROS Immunodiagnostic Products Total β-hCG II Reagent
Pack Traditional 510(k)

General Device Characteristic Differences:

Assay Principle	Sandwich immunoassay	Sandwich immunoassay.In the modified VITROS ImmunodiagnosticProducts Total β-hCG II reagent pack, the mouseanti-β hCG antibody has been removed from theBiotin Reagent and coated directly onto the well. Themodification is to allow the biotinylated antibodycapture conjugate to be pre- bound to the well,eliminating the risk of biotin interference.
Reagent pack	Biotin-conjugated mouse monoclonal anti-β-hCGcoated microwell	Biotin-conjugated mouse monoclonal anti-β-hCGreagent buffer
Instrumentation	VITROS 5600 Integrated System	VITROS ECi/ECiQ/3600 Immunodiagnostic Systemsand the VITROS 5600/XT 7600 Integrated Systems

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

Nonclinical Performance

Several nonclinical tests were performed.

Stability Studies

Long term stability and on-board storage performance was evaluated consistent with methods based on CLSI EP25-A.

Long Term Stability: Four runs have been performed on each of 3 Lots at each time-point, monthly intervals, data supports a 52 week shelf-life.

On-board Stability: Three Lots of the VITROS Immunodiagnostic Products Total ß-hCG II reagent pack will be stored opened refrigerated for up to 12 weeks. Four runs were performed on each Lot at each time-point for fresh and open, all results were acceptable and support a claim of 6 weeks on-board stability currently.

Precision

Precision was evaluated consistent with CLSI document EP05-A3, Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline-Third Edition. Six (6) precision fluids, covering the analytical measuring interval, were evaluated for performance. For one reagent Lot, two (2) replicates of each precision fluid were run on two (2) occasions per day for twenty (20) days, for a total of 80 data points per fluid.

The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of test results.

	Units = mIU/mL (IU/L)
System	Mean hCG Conc.	Repeatability*		Within Lab**		No. ofObs.	No. ofDays
		SD	%CV	SD	%CV
5600	5.33	0.095	1.80%	0.281	5.30%	80	20
	15.46	0.181	1.20%	0.706	4.60%	80	20
	45.67	0.666	1.50%	1.560	3.40%	80	20
	297.34	4.970	1.70%	8.640	2.90%	80	20
	4708.50	82.29	1.70%	151.22	3.20%	80	20
	9651.00	286.71	3.00%	464.48	4.80%	80	20

Product Claim

Alternate Units (IU/L) are equivalent to Conventional Units (mIU/ml).

**Repeatability (formerly called within-run precision) was determined using two replicates per run.

*** Within Lab precision was determined using a single reagent lot and a single calibration.

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

Additional Precision Analysis Summary

Sample	Mean	With-in Run		Between Run		Within Day		Between Day		Between Lot		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
PP1	5.33	0.095	1.8%	0.190	3.6%	0.213	4.0%	0.183	3.4%	0.201	3.7%	0.281	5.3%
PP2	15.46	0.181	1.2%	0.647	4.2%	0.672	4.3%	0.215	1.4%	0.706	4.4%	0.706	4.6%
PP3	45.67	0.666	1.5%	1.260	2.8%	1.425	3.1%	0.632	1.4%	1.610	3.5%	1.559	3.4%
PP4	297.34	4.967	1.7%	4.479	1.5%	6.688	2.2%	5.463	1.8%	12.627	4.2%	8.635	2.9%
PP5	4708.5	82.29	1.7%	95.10	2.0%	125.76	2.7%	83.98	1.8%	268.80	5.7%	151.22	3.2%
PP6	9651.0	286.71	3.0%	230.87	2.4%	368.11	3.8%	283.26	2.9%	610.09	6.4%	464.48	4.8%

Units = mIU/mL (IU/L)

Multiple System (Reproducibility) Study Summary

Six precision fluids (RP1-6) made by pooling female serum samples were evaluated for performance according to CLSI guideline EP05-A3. Five (5) replicates of each fluid were run on one (1) occasion per day for five (5) days The samples were run in randomized order on three (3) VITROS 5600 integrated systems.

		Within-Run		Between-Day		Within-Site		Between-Site		Total
Sample	Mean (mIU/mL)	SD	% CV	SD	% CV	SD	% CV	SD	% CV	SD	%CV
RP1	4.38	0.10	2.3%	0.14	3.2%	0.17	3.9%	0.37	8.5%	0.41	9.4%
RP2	15.00	0.23	1.5%	0.42	2.8%	0.48	3.2%	0.90	6.0%	1.02	6.8%
RP3	49.55	0.77	1.6%	0.94	1.9%	1.21	2.5%	1.96	4.0%	2.31	4.7%
RP4	306.35	5.11	1.7%	4.50	1.5%	6.81	2.2%	7.19	2.3%	9.90	3.2%
RP5	4781.1	105.94	2.2%	89.54	1.9%	138.70	2.9%	92.27	1.9%	166.59	3.5%
RP6	8329.9	249.62	3.0%	219.68	2.6%	332.52	4.0%	220.88	2.7%	399.20	4.8%

Detection Capability

Detection studies for the VITROS Immunodiagnostic Products Total β-hCG II reagent pack were evaluated consistent with CLSI document EP17-A2, Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second Edition. Four serums containing no measurable human chorionic gonadotropin (hCG) were used for determining the LoB. The study design was 2 replicates per run, 2 runs per day over 5 test days = 20 reps per test fluid x 4 fluids = 80 replicates x 3 lots = 240 total replicates. The representative LoB is 0.00 mIU/mL (IU/L) which supports the claimed LOB of 0.05mIU/mL.

Five samples were used for establishing the LoD, which were targeted at 1 to 5 times the LoB concentration. The LoD samples were neat serum samples containing low levels of endogenous hCG to achieve the approximate target hCG concentrations. The LoD serums were also used to analyze the LoQ via the Precision Profile method. The Total Error Analysis for LoQ was determined by testing a separate set of four LoQ fluids which were then analyzed to calculate the LoQ via the Total Error method. All samples were run using three reagent lots on one VITROS 5600 System, 6 replicates per run, 2 runs per day over 5 test days = 60 reps per fluid x 5 fluids = 300 replicates x 3 lots = 900 total replicates.

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

VITROS Total ß-hCG II Reagent Pack Traditional 510(k)

The representative Limit of Detection (LoD) for the VITROS Immunodiagnostic Products Total B-hCG II reagent pack is 0.21 mIU/mL (IU/L) determined consistent with CLSI document EP17. This supports the claimed LoD of 0.70 mIU/mL (IU/L). The Limit of Quantitation (LoQ) for the VITROS Immunodiagnostic Products Total 8hCG II reagent pack was designed to be less than or equal to the currently claimed low end of the measuring range of 2.39 mIU/mL (IU/L) at 20% CV. The representative LoQusing the Total Error approach was determined to be 2.32 mIU/mL (IU/L), consistent with CLSI document EP17. The claimed LoQ has been verified at 2.39 mIU/mL (IU/L).

Linearity

Linearity was established in accordance with the CLSI guideline EP06 2nd edition. In a study with two series, one across the entire range (pools 1a to 10a) and one in the lower range of the assay (pools 1b to 10b). 10 replicates of pools 1a/16 and 10a/10b, and 5 replicates of pools 2a/2b to 9a/9b were run on one VITROS 5600 Integrated System. Results support linearity from 1.51 mIU/mL (IU/L) to 15695 mIU/mL (IU/L).

				Slope		Intercept
Lot	DilutionRange	% Recovery	Estimate	95% CI	Estimate	95% CI	R2
2643	38.6 to 15695	92.2% to108.2%	0.991	0.949 to1.033	0.10	-2.56 to2.76	0.996
2643	1.51 to 51.18	99.9% to112.7%	1.025	0.995 to1.056	0.07	-0.09 to0.22	0.999

Measuring Range

VITROS System	Measuring (Reportable) Range
5600	2.39*-15,000 mIU/mL(IU/L)

Lower limit of measuring range reported by the system software is based on the Limit of Quantitation.

Matrix Comparison

Serum and plasma (Lithium-Heparin and K2-EDTA) specimen matrices were determined to be equivalent. The results met the acceptance criteria for the comparison between serum and plasma (Lithium-Heparin and K2-EDTA) specimens spanning the expected measuring interval. Based on the analysis, serum and plasma (Lithium-Heparin and K2-EDTA) are suitable specimen matrices for use with the VITROS Immunodiagnostic Products Total ß-hCG II reagent pack.

Specimens Recommended

Serum
. Lithium-HeparinPlasma
K2- EDTA Plasma ●

Specimens Not Recommended

Do not use turbid specimens. Turbidity in specimens may affect test results. .

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

VITROS 5600
Weighted DemingRegression	Li-Hep Plasma	K2-EDTA Plasma
Slope	0.978	0.978
Corr. Coef r	0.999	0.999
n	41	41

Analytical Specificity

Known Interferents

The VITROS Immunodiagnostic Products Total ß-hCG II reagent pack was screened for interfering substances at hCG concentrations of approximately 5.00 mIU/mL (IU/L), 50.00 mIU/mL (IU/L), and 10,000 mIU/mL (IU/L) following CLSI EP07 and EP37. Of the compounds tested, none was found to cause a bias of >10%.

For substances that were tested and did not interfere, refer to "Substances that do not Interfere."

Substances that do not Interfere

The substances listed in the table below were tested with the VITROS Immunodiagnostic Products Total ß-Hcg II reagent pack following CLSI EP07and EP37 and found not to cause bias > 10% at Hcg concentrations of approximately 5.00 mIU/mL (IU/L), 50.00 mIU/mL (IU/L), and 10,000 mIU/mL (IU/L) at the test concentrations shown.

Substance	Concentration	Concentration
Acetaminophen	15.6 mg/Dl	1032 µmol/L
N-Acetylcysteine	15.0 mg/Dl	920 µmol/L
Alpha-tocopherol	6.45 mg/Dl	150 µmol/L
Amoxicillin	5.40 mg/Dl	148 µmol/L
Ascorbic acid	5.25 mg/Dl	298 µmol/L
Bilirubin, conjugated	40 mg/Dl	475 µmol/L
Bilirubin, unconjugated	40 mg/Dl	475 µmol/L
Biotin	0.351 mg/Dl	14.3 µmol/L
Carbamazepine	4.50 mg/Dl	191 µmol/L
Cefoxitin sodium	695 mg/Dl	15.5 mmol/L
Cholesterol	400 mg/Dl	10.3 mmol/L
Codeine	0.141 mg/Dl	5 µmol/L
Colecalciferol	19.2 µg/Dl	0.5 µmol/L
Cotinine	0.24 mg/Dl	13.6 µmol/L
Dextran 40	2400 mg/Dl	600 µmol/L
Dextromethorphan	0.00156 mg/Dl	0.042 µmol/L
Dipyrone	100 mg/Dl	N/A
Enoxaparin	360 U/DI	N/A
Ethanol	600 mg/Dl	130 mmol/L

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

Substance		Concentration
Furosemide	1.59 mg/Dl	48 µmol/L
Gamma globulin	6 g/Dl	N/A
HAMA (Human Anti-MouseAntibodies)	800 µg/L	0.005 µmol/L
Hemoglobin	1000 mg/Dl	155 µmol/L
Hydralazine hydrochloride	1.44 mg/Dl	73.2 µmol/L
Hydrocodone	0.0072 mg/Dl	0.2 µmol/L
Ibuprofen	21.9 mg/Dl	1.06 mmol/L
Intralipid	2000 mg/Dl	19.2 mmol/L
Levothyroxine	0.0429 mg/Dl	0.552 µmol/L
Loratadine	0.0087 mg/Dl	0.227 µmol/L
Naproxen	36.0 mg/Dl	1.43 mmol/L
Nifedipine	0.0588 mg/Dl	1.7 µmol/L
Nitrofurantoin	0.213 mg/Dl	8.94 µmol/L
Omeprazole	0.840 mg/Dl	24.3 µmol/L
Phenytoin	6.00 mg/Dl	238 µmol/L
Prednisone	0.010 mg/Dl	0.280 µmol/L
Propranolol HCl	0.115 mg/Dl	3.89 µmol/L
Rheumatoid factor	900 IU/mL	N/A
Salicylic acid	2.86 mg/Dl	207 mmol/L
Sodium azide	100 mg/Dl	15.4 mmol/L
Sulfamethoxazole	40 mg/Dl	1.58 mmol/L
Theophylline	6.0 mg/Dl	333 µmol/L
Total Protein	15 g/dL	17.1 nmol/L
Triglycerides	1500 mg/Dl	16.9 mmol/L
Trimethoprim	4.2 mg/Dl	145 µmol/L
Vancomycin hydrochloride	12.3 mg/Dl	8.28 mmol/L

Cross-Reactivity

The cross-reactivity of the VITROS Immunodiagnostic Products Total β-hCG II reagent pack was evaluated by adding the following test substances to a sample containing no hCG.

Substance	TestedConcentration	VITROS 5600 -% CrossReactivity
FSH	400 mIU/mL	ND
LH	400 mIU/mL	ND
TSH	200 mIU/mL	ND

% Cross-Reactivity Summary with a hCG Negative Sample

*Not Detectable (ND). Concentration was below the measuring interval of the test, 2.39 to 15,000 mIU/mL (IU/L). Ortho Clinical Diagnostics

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

Each potential cross-reactant was spiked into hCG pools at hCG concentrations of approximately 25 mIU/mL Test substances were spiked at a volume that constituted no more than 5% of the volume of the test sample.

Substance	Tested Concentration	% Cross Reactivity
FSH	400 mIU/mL	0.6%
LH	400 mIU/mL	-0.3%
TSH	200 mIU/mL	0.4%

% Cross-Reactivity Summary with a Pool at hCG Concentrations of Approximately 25 mIU/mL.

Dilution

Samples with concentrations greater than the measuring range may be diluted automatically on the system up to 400-fold (1 part sample with 399 parts diluent) by the VITROS Integrated System with VITROS High Sample Diluent B Reagent Pack prior to test. Refer to the High Sample Diluent B Reagent Pack Instructions for Use.

Expected Values

Adult Reference Interval

The adult reference interval was validated following CLSI document EP28-A3c Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory: Approved Guideline — Third Edition.

The distribution of hCG values from 180 normal healthy non-pregnant blood donors showed equivalency to the expected values claim published in the Instructions for Use of the current VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack, therefore the current distribution of results for healthy non-pregnant subjects will be transferred to the updated VITROS Immunodiagnostic Products Total ß-hCG II Reagent Pack.

Product Claim:

		Units = mIU/mL (IU/L)
Sample type	Numberofsamples	Mean	Min	Max	2.5thPercentile	97.5thPercentile
Normal Male	98	0.02	0.00	1.06	0.00	0.08
Normal Female	123	0.46	0.00	5.42	0.00	4.32
Post menopausal	69	1.52	0.00	6.66	0.00	6.46
Total	290	0.56	0.00	6.66	0.00	4.83

. As a guide, the following ranges were determined. Concentrations of hCG measured in samples from apparently healthy, non-Ortho Clinical Diagnostics

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

Sample Type	Number of Samples	Units (mIU/mL)
		Mean	Min	Max	2.5th Percentile	97.5th Percentile
Normal Male	60	0.00	0.00	0.04	0.00	0.04
Normal Female	60	0.07	0.00	2.78	0.00	1.44
Post- Menopausal	60	1.32	0.00	5.61	0.00	5.08
Total	180	0.47	0.00	5.61	0.00	3.73

Distribution of Healthy Subjects Results for the updated VITROS Immunodiagnostic Products Total B-hCG II reagent pack:

Traceability of Calibration

Calibration of the VITROS Immunodiagnostic Products Total ß-hCG II reagent pack is traceable to inhouse reference calibrators, which have been value-assigned with reference to the 4th International Standard (NIBSC 75/589).

Method Comparison

Accuracy was evaluated consistent with CLSI guideline EP09c Measurement Procedure Comparison and Bias Estimation Using Patient Samples. 3rd ed. Human serum samples were obtained from certified vendors and tested neat. A total of 135 samples analyzed. The samples were tested in singleton using one reagent lot and one VITROS 5600 Integrated System.

The plots and table show the results of a method comparison study using patient serum samples analyzed on the VITROS Immunodiagnostic Products Total β-hCG II reagent pack compared with those analyzed on the VITROS Immunodiagnostic Products Total {}-hCG II reagent pack (GEM.1076) on the VITROS 5600 Integrated System. The relationship between the 2 methods was determined by Weighted Deming regression.

Image /page/12/Figure/8 description: This image shows a scatter plot comparing two methods of measuring VITROS Total B-hCG. The x-axis represents the comparative method, VITROS Total B-hCG (GEM.1076) in mIU/mL, ranging from 0 to 15000. The y-axis represents VITROS Total B-hCG II (GEM.1076A) in mIU/mL, also ranging from 0 to 15000, with a red line indicating the line of best fit and dashed lines indicating the confidence interval.

Image /page/12/Figure/9 description: This image shows a scatter plot comparing two methods of measuring VITROS Total B-hCG. The x-axis represents the comparative method, VITROS Total B-hCG (GEM.1076) in IU/L, while the y-axis represents VITROS Total B-hCG II (GEM.1076A) in IU/L. The plot includes a red line representing the line of equality and two dashed lines indicating the range of acceptable agreement. The data points are clustered tightly around the line of equality, indicating strong agreement between the two methods across a range of values from 0 to 15000 IU/L.

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

System	n	Slope(95% CI)	CorrelationCoefficient	Conventional Units(mIU/mL)		Alternate Units (IU/L)
				Range ofSamples	Intercept(95% CI)	Range ofSamples	Intercept(95% CI)
5600 vs.ComparativeMethod	135	0.99(0.977 to0.995)	0.998	2.78-14,675	-0.0215(-0.160 to0.117)	2.78-14,675	-0.0215(-0.160 to0.117)

Conclusion

The conclusions drawn from the nonclinical tests (discussed above) demonstrate the updated VITROS Immunodiagnostic Products Total β-hCG II reagent pack is as safe, effective, and performs as well as the cleared predicate device. The information submitted in the premarket notification is complete and supports a substantial equivalence decision.

Regulation Number and Section

§ 862.1155 Human chorionic gonadotropin (HCG) test system.

(a)
Human chorionic gonadotropin (HCG) test system intended for the early detection of pregnancy —(1)Identification. A human chorionic gonadotropin (HCG) test system is a device intended for the early detection of pregnancy is intended to measure HCG, a placental hormone, in plasma or urine.(2)
Classification. Class II.(b)
Human chorionic gonadotropin (HCG) test system intended for any uses other than early detection of pregnancy —(1)Identification. A human chorionic goadotropin (HCG) test system is a device intended for any uses other than early detection of pregnancy (such as an aid in the diagnosis, prognosis, and management of treatment of persons with certain tumors or carcinomas) is intended to measure HCG, a placental hormone, in plasma or urine.(2)
Classification. Class III.(3)
Date PMA or notice of completion of a PDP is required. As of the enactment date of the amendments, May 28, 1976, an approval under section 515 of the act is required before the device described in paragraph (b)(1) may be commercially distributed. See § 862.3.