VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator

DEN210038 · Ortho-Clinical Diagnostics, Inc. · QVP · May 5, 2023 · Microbiology

Device Facts

• Record ID: DEN210038
• Device Name: VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator
• Applicant: Ortho-Clinical Diagnostics, Inc.
• Product Code: QVP · Microbiology
• Decision Date: May 5, 2023
• Decision: DENG
• Submission Type: Direct
• Regulation: 21 CFR 866.3983
• Device Class: Class 2

Indications for Use

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack when used in combination with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator is a chemiluminescent immunoassay intended for the qualitative detection of IgG antibodies to SARS-CoV-2 in human serum and plasma (K2-EDTA and K3-EDTA) samples collected on or after 15 days post-symptom onset using the VITROS ECi/ECiQ/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems. The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is intended for use as an aid in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection.

Device Story

The VITROS Anti-SARS-CoV-2 IgG test is a chemiluminescent immunoassay for laboratory professional use on VITROS automated systems. It detects IgG antibodies in human serum or plasma (K2/K3-EDTA). The test uses a two-stage immunometric reaction: patient antibodies bind to SARS-CoV-2 spike protein-coated wells; unbound sample is washed away; HRP-labeled anti-human IgG conjugate is added and binds to the complex. A luminogenic substrate is added; HRP catalyzes light emission, which is measured by the system. Results are calculated as a normalized signal (S/C) relative to a cutoff. The output aids clinicians in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection. It supports infection control and surveillance efforts.

Clinical Evidence

No clinical data provided in the document.

Technological Characteristics

Chemiluminescent immunoassay; utilizes VITROS ECi/ECiQ/3600 and 5600/XT 7600 systems. Reagents include Reagent Pack and Calibrator. Qualitative detection of IgG antibodies. Automated processing. Requires specific specimen types (serum, K2-EDTA/K3-EDTA plasma).

Indications for Use

Indicated for qualitative detection of IgG antibodies to SARS-CoV-2 in human serum and plasma (K2-EDTA, K3-EDTA) from individuals 15+ days post-symptom onset. Aid in identifying adaptive immune response indicating recent/prior infection. Not for acute infection diagnosis or donor screening.

Regulatory Classification

Identification

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is a qualitative chemiluminescent immunoassay intended for the detection of IgG antibodies to SARS-CoV-2 in human serum and plasma (K2-EDTA and K3-EDTA). It is performed on the VITROS ECi/ECiQ/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems. The test is intended for use as an aid in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection.

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food & Drug Administration (FDA). The logo consists of two parts: a symbol on the left and the text "FDA U.S. FOOD & DRUG ADMINISTRATION" on the right. The symbol features a stylized image of an eagle, while the text is written in blue, with "FDA" in a larger, bolder font than the rest of the text.

# EVALUATION OF AUTOMATIC CLASS III DESIGNATION FOR VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator DECISION SUMMARY

#### Background Information: I

### A De Novo Number

DEN210038

### B Applicant

Ortho-Clinical Diagnostics, Inc.

### C Proprietary and Established Names

VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator

#### D Regulatory Information

| Product<br>Code(s) | Classification | Regulation<br>Section | Panel |
|--------------------|----------------|------------------------------------------------|-------------------|
| QVP | Class II | 21 CFR 866.3983 - SARS-<br>CoV-2 serology test | MI - Microbiology |

#### II Submission/Device Overview:

### A Purpose for Submission:

De Novo request for evaluation of automatic class II designation for the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test comprised of (1) the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, (2) the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator, and (3) the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Controls.

#### B Measurand:

IgG antibodies to SARS-CoV-2 in human serum, K2-EDTA plasma, and K3-EDTA plasma.

### C Type of Test:

Chemiluminescent Immunoassay.

#### III Indications for Use:

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### A Indication(s) for Use: Rx ONLY

For in vitro diagnostic and laboratory professional use.

# VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack when used in combination with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator is a chemiluminescent immunoassay intended for the qualitative detection of IgG antibodies to SARS-CoV-2 in human serum and plasma (K2-EDTA and K3-EDTA) samples collected on or after 15 days post-symptom onset using the VITROS ECi/ECiO/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems. The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is intended for use as an aid in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection.

VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator

For use in the calibration of the VITROS ECi/ECiQ/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems for the in vitro qualitative detection of IgG antibodies to SARS-CoV-2 in human serum and plasma.

# B Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

### C Special Instrument Requirements:

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack when used in combination with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrators uses the following VITROS Systems (instruments):

- VITROS ECi/ECiQ Immunodiagnostic Systems .
- VITROS 3600 Immunodiagnostic Systems .
- . VITROS 5600 Integrated Systems
- . VITROS XT 7600 Integrated Systems

These VITROS systems were cleared previously as part of premarket notifications: K962919, K083173, K081543, and K182063 respectively,

#### IV Device/System Characteristics:

### A Device Description:

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is a qualitative chemiluminescent immunoassay performed on the VITROS Systems (VITROS ECi/ECiO Immunodiagnostic System, VITROS 3600 Immunodiagnostic System, VITROS 5600 Integrated

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System and VITROS XT 7600 Integrated System) providing fully automated random-access testing.

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is performed using the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack in combination with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator and the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Controls on the VITROS Systems.

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack is supplied as ready to use and contains:

- . 100 wells coated with 100ng/well of recombinant SARS-CoV-2 spike antigen derived from human cells.
- 18.0 mL assay reagent (buffer with bovine protein stabilizers and antimicrobial agent) .
- . 20.4 mL conjugate reagent [anti-human IgG (murine monoclonal) conjugated to horseradish peroxidase, 5ng/mL] in buffer with bovine protein stabilizers and antimicrobial agent.

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator contains:

- 2 vials of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 1gG Calibrator 0 (anti-SARS-CoV-2 IgG in anti-SARS-CoV-2 IgG negative human serum with antimicrobial agent, 1 mL)
- Lot calibration card .
- . Protocol card
- . 8 calibrator bar code labels

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Controls contain:

- 3 sets of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Controls 1 and 2 . (defibrinated human plasma with anti-microbial agent, 2 mL). Control 1 is non-reactive and Control 2 is reactive
The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is designed for use on the VITROS Systems. The VITROS Systems use the following ancillary reagents (general purpose reagents):

- . VITROS Immunodiagnostic Products Signal Reagent
- VITROS Immunodiagnostic Products Universal Wash Reagent .

# B Principle of Operation

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test is performed using the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack and the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator on the VITROS ECi/ECiO/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems. An immunometric technique is used; this involves a two- stage reaction. In the first stage antibodies to SARS-CoV-2 present in the sample bind with SARS-CoV-2 spike protein coated on the well. Unbound sample is removed by washing. In the second stage horseradish peroxidase (HRP)-labeled murine monoclonal anti-human IgG antibodies are

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added in the conjugate reagent. The conjugate binds specifically to the antibody portion of the antigen-antibody complex. If complexes are not present, the unbound conjugate is removed by the subsequent wash step.

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.

| Test Type | System | Incubation Time | Time to first result | Test Temperature | Reaction Sample<br>Volume |
|--------------|----------------------------------|-----------------|----------------------|------------------|---------------------------|
| Immunometric | ECi/ECiQ. 3600.<br>5600. XT 7600 | 37 minutes | 48 minutes | 37 ℃ | 20 ul |

* Not all products and systems are available in all countries.

# Figure 1: Reaction Scheme

Image /page/3/Figure/5 description: The image shows a diagram of a process involving several components. It starts with a recombinant SARS-CoV-2 antigen, followed by Anti-SARS-CoV-2, and then HRP-labeled anti-human IgG murine monoclonal. The process culminates in luminescence, which is achieved using a signal reagent with an enhancer.

# C Instrument Description Information

### 1. Instrument Name:

VITROS Systems (instruments):

- . VITROS ECi/ECiQ Immunodiagnostic System
- . VITROS 3600 Immunodiagnostic System
- VITROS 5600 Integrated System .
- VITROS XT 7600 Integrated System. .
- 2. Specimen Identification:

Not applicable

- 3. Specimen Sampling and Handling:
#### Specimen Sampling:

The specimens recommended for this assay are:

- . Serum
- K2-EDTA Plasma
- K3-EDTA Plasma

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# Specimen Handling and Storage:

- . Specimens may be stored for up to 24 hours at room temperature (15-30℃) or 7 days at 2 - 8ºC.
- Specimens may be stored frozen at < 20°C for <4 weeks. .
- Specimens may be subjected to up to one freeze-thaw cvcles. .

# 4. Calibration:

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator is provided together with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack.

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator contains anti-SARS-CoV-2 IgG in human serum. In addition, the calibrator contains the lot calibration card, protocol card and 8 calibrator bar code labels.

Calibration is lot-specific: reagent packs and calibrators are linked by lot number. Reagent packs from the same lot may use the same calibration. The calibrator is supplied frozen. The analyzer automatically processes the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Calibrator in duplicate. Results are calculated as a normalized signal, relative to a cutoff value. During the calibration process a lot-specific parameter is used to determine a valid stored cutoff value for the VITROS Immunodiagnostic and VITROS Integrated Systems.

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test should be calibrated:

- . When the reagent pack and calibrator lot changes
- . Every 28 days
- After specified service procedures have been performed .
- If quality control results are consistently outside of the acceptable range. .

# 5. Quality Control:

The VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Controls are provided separately from the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG Reagent Pack, and contains:

- · Control I: Anti-SARS-CoV-2 IgG non-reactive human serum or defibrinated plasma.
- . Control 2: Anti-SARS-CoV-2 IgG reactive human serum or defibrinated plasma.

# Quality Control Procedure

- To verify system performance, analyze control materials: .
- O After calibration
- o If the system is turned off for more than 2 hours
- · After reloading reagent packs that have been removed from the MicroWell Supply and stored for later use
- According to local regulations or at least once each day that the test is performed

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- · After specified service procedures are performed
- . If controls results fall outside the acceptance range, investigate the cause before deciding whether to report patient results.

#### V Standards/Guidance Documents Referenced:

CLSI EP05-A3 Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition

CLSI EP07 Interference testing in Clinical Chemistry

- CLSI EP09 Measurement Procedure Comparison and Bias Estimation Using Patient Samples. Third Edition (2016).
- CLSI EP35 Assessment of Equivalence or Suitability of Specimen Type for Medical Laboratory measurement Procedure

CLSI EP37 Supplemental Tables for Interference Testing in Clinical Chemistry

#### Performance Characteristics: VI

### A Analytical Performance:

#### 1. Precision/Reproducibility:

A. Within-Laboratory Precision: The within laboratory precision of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test was evaluated with EDTA plasma pools samples (PP4, PP5, and PP6), negative and positive quality control materials (PP1, PP2), and the calibrator (PP3) on the VITROS ECi/ECiQ and 3600 Immunodiagnostics Systems and the VITROS 5600 and XT7600 Integrated Systems following the CLSI document EP05-A3. A total of 3 lots of reagent packs, calibrators and controls were included in the study. For each reagent lot, operators ran two replicates of each precision panel sample on two occasions per day for twenty non-consecutive days.

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.

Each precision panel was tested in duplicate, in two runs per day, on 20 non-consecutive days, on each instrument for a total of 80 observations per sample and lot. The study included three reagent lots and was evaluated within a single calibration cycle (2 replicates x 2 runs x 20 days x 3 lots= 240 observations total). The VITROS Anti-SARS-CoV-2 IgG's total precision %CV (from the "total" standard deviation) for the S/C values ranged from 3.4% -32.2%, depending upon the sample and instrument.

| VITROS<br>System | Panel<br>member | Number<br>of<br>Observ. | Grand<br>Mean<br>(S/C) | Repeatability<br>(Within Run) | | Between-Run | | Between-Day | | Between-Lot | | Within-<br>Laboratory | |
|------------------|-----------------|-------------------------|------------------------|-------------------------------|-----|-------------|-----|-------------|-----|-------------|-----|-----------------------|-----|
| ECi/<br>ECiQ | PP1 | 240 | 0.00 | 0.000 | N/A | 0.000 | N/A | 0.000 | N/A | 0.001 | N/A | 0.001 | N/A |
| ECi/<br>ECiQ | PP2 | 240 | 0.92 | 0.018 | 2.0 | 0.024 | 2.6 | 0.034 | 3.7 | 0.030 | 3.3 | 0.054 | 5.9 |

#### Table 1. Within-Laboratory Precision Study Data Summary (for all 4 VITROS analyzers).

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| | PP3 | 240 | 0.25 | 0.007 | N/A | 0.011 | N/A | 0.011 | N/A | 0.013 | N/A | 0.022 | N/A |
|---------|-----|-----|------|-------|-----|-------|-----|-------|-----|-------|------|-------|------|
| | PP4 | 240 | 0.94 | 0.027 | 2.9 | 0.032 | 3.4 | 0.033 | 3.5 | 0.091 | 9.7 | 0.106 | 11.3 |
| | PP5 | 240 | 3.25 | 0.086 | 2.6 | 0.062 | 1.9 | 0.084 | 2.6 | 0.169 | 5.2 | 0.216 | 6.6 |
| | PP6 | 240 | 7.62 | 0.063 | 0.8 | 0.076 | 1.0 | 0.112 | 1.5 | 0.208 | 2.7 | 0.257 | 3.4 |
| | PP1 | 240 | 0.00 | 0.000 | N/A | 0.000 | N/A | 0.001 | N/A | 0.002 | N/A | 0.002 | N/A |
| | PP2 | 240 | 1.18 | 0.024 | 2.0 | 0.025 | 2.1 | 0.021 | 1.8 | 0.240 | 20.3 | 0.243 | 20.6 |
| | PP3 | 240 | 0.32 | 0.007 | N/A | 0.008 | N/A | 0.006 | N/A | 0.078 | N/A | 0.079 | N/A |
| 3600 | PP4 | 240 | 1.18 | 0.028 | 2.4 | 0.034 | 2.9 | 0.033 | 2.8 | 0.376 | 31.9 | 0.380 | 32.2 |
| | PP5 | 240 | 3.94 | 0.083 | 2.1 | 0.092 | 2.3 | 0.072 | 1.8 | 1.064 | 27.0 | 1.073 | 27.2 |
| | PP6 | 240 | 8.56 | 0.099 | 1.2 | 0.099 | 1.2 | 0.105 | 1.2 | 1.610 | 18.8 | 1.619 | 18.9 |
| | PP1 | 240 | 0.00 | 0.000 | N/A | 0.000 | N/A | 0.000 | N/A | 0.001 | N/A | 0.001 | N/A |
| | PP2 | 240 | 1.03 | 0.020 | 1.9 | 0.027 | 2.6 | 0.027 | 2.6 | 0.058 | 5.6 | 0.072 | 7.0 |
| | PP3 | 240 | 0.28 | 0.006 | N/A | 0.010 | N/A | 0.007 | N/A | 0.025 | N/A | 0.028 | N/A |
| 5600 | PP4 | 240 | 1.02 | 0.026 | 2.5 | 0.036 | 3.5 | 0.026 | 2.5 | 0.140 | 13.7 | 0.149 | 14.6 |
| | PP5 | 240 | 3.43 | 0.073 | 2.1 | 0.074 | 2.2 | 0.095 | 2.8 | 0.288 | 8.4 | 0.321 | 9.4 |
| | PP6 | 240 | 7.51 | 0.097 | 1.3 | 0.105 | 1.4 | 0.119 | 1.6 | 0.276 | 3.7 | 0.333 | 4.4 |
| | PP1 | 240 | 0.00 | 0.000 | N/A | 0.000 | N/A | 0.000 | N/A | 0.001 | N/A | 0.001 | N/A |
| | PP2 | 240 | 1.21 | 0.043 | 3.6 | 0.042 | 3.5 | 0.016 | 1.3 | 0.153 | 12.6 | 0.165 | 13.6 |
| | PP3 | 240 | 0.32 | 0.009 | N/A | 0.009 | N/A | 0.010 | N/A | 0.052 | N/A | 0.055 | N/A |
| XT 7600 | PP4 | 240 | 1.19 | 0.034 | 2.9 | 0.042 | 3.5 | 0.037 | 3.1 | 0.239 | 20.1 | 0.248 | 20.8 |
| | PP5 | 240 | 3.94 | 0.078 | 2.0 | 0.093 | 2.4 | 0.088 | 2.2 | 0.604 | 15.3 | 0.622 | 15.8 |
| | PP6 | 240 | 8.46 | 0.113 | 1.3 | 0.117 | 1.4 | 0.098 | 1.2 | 0.896 | 10.6 | 0.916 | 10.8 |

N/A: Not Applicable, %CV is not meaningful for S/C results <0.5.

B. Reproducibility (Between-Laboratory Precision): The reproducibility of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test was determined with EDTA plasma pools samples (GRP4, GRP5, and GRP6), quality controls (GRP1 and GRP2) and customer calibrator (GRP3) using the VITROS ECi/ECiQ and 3600 Immunodiagnostics Systems, and the VITROS 5600 and XT7600 Integrated Systems. Samples were measured in triplicate using 3 reagent lots, in 2 runs per day over 5 days at 3 sites, according to CLSI EP05-A3. Variance components were calculated. The VITROS Anti-SARS-CoV-2 IgG test total reproducibility %CV (from the "total" standard deviation) for the S/C values ranged from 6.1% - 22.8%, depending upon the sample and instrument. The following results were observed per analyzer:

Table 2. Reproducibility of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test evaluated on VITROS ECi/ECiO Immunodiagnostic Systems

| Panel<br>Member<br>r | Number<br>of<br>Observ. | Grand<br>Mean<br>(S/C) | Repeatability<br>(Within Run) | | Between Run | | Between Day | | Between Lota | | Within-<br>Laboratoryb | | Between-Sitec | | Reproducibility<br>(Overall)d | |
|----------------------|-------------------------|------------------------|-------------------------------|-----------|-------------|-----------|-------------|-----------|--------------|-----------|------------------------|-----------|---------------|-----------|-------------------------------|-----------|
| | | | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) |
| GRP1 | 270 | 0.01 | 0.002 | N/A+ | 0.000 | N/A+ | 0.000 | N/A+ | 0.003 | N/A+ | 0.003 | N/A+ | 0.003 | N/A+ | 0.006 | N/A+ |
| GRP2 | 270 | 0.98 | 0.058 | 5.9 | 0.000 | 0.0 | 0.021 | 2.2 | 0.000 | 0.0 | 0.062 | 6.3 | 0.018 | 1.8 | 0.095 | 9.6 |
| GRP3 | 270 | 0.28 | 0.016 | N/A+ | 0.007 | N/A+ | 0.010 | N/A+ | 0.021 | N/A+ | 0.029 | N/A+ | 0.000 | N/A+ | 0.034 | N/A+ |
| GRP4 | 270 | 1.08 | 0.059 | 5.5 | 0.011 | 1.0 | 0.035 | 3.2 | 0.206 | 19.2 | 0.217 | 20.2 | 0.000 | 0.0 | 0.225 | 21.0 |
| GRP5 | 270 | 3.59 | 0.164 | 4.6 | 0.040 | 1.1 | 0.071 | 2.0 | 0.479 | 13.3 | 0.513 | 14.3 | 0.024 | 0.7 | 0.545 | 15.2 |
| GRP6 | 270 | 7.84 | 0.220 | 2.8 | 0.000 | 0.0 | 0.091 | 1.2 | 0.425 | 5.4 | 0.487 | 6.2 | 0.311 | 4.0 | 0.665 | 8.5 |

11 Between lot: Variability of the test performance from lot to lot.

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b Within-Laboratory variability contains the Within Run, Between Run, Between Day and Between Lot variance components.

Between site: Variability of the test performance from site to site.

d Reproducibility: Variability of the test incorporating factors of site, lot, run and day.

+ % CVs are not meaningful for S/C results < 0.50.

| PanelMember | Number of Observ. | Grand Mean (S/C) | Repeatability (Within Run) | | Between Run | | Between Day | | Between Lota | | Within-Laboratoryb | | Between-Sitec | | Reproducibility (Overall)d | |
|-------------|-------------------|------------------|----------------------------|--------|-------------|--------|-------------|--------|--------------|--------|--------------------|--------|---------------|--------|----------------------------|--------|
| | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| GRP1 | 270 | 0.00 | 0.001 | N/A+ | 0.000 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.002 | N/A+ | 0.001 | N/A+ | 0.002 | N/A+ |
| GRP2 | 270 | 1.05 | 0.046 | 4.4 | 0.000 | 0.0 | 0.029 | 2.8 | 0.051 | 4.9 | 0.074 | 7.1 | 0.010 | 1.0 | 0.089 | 8.5 |
| GRP3 | 270 | 0.29 | 0.012 | N/A+ | 0.003 | N/A+ | 0.012 | N/A+ | 0.026 | N/A+ | 0.032 | N/A+ | 0.008 | N/A+ | 0.038 | N/A+ |
| GRP4 | 270 | 1.08 | 0.043 | 4.0 | 0.012 | 1.1 | 0.017 | 1.5 | 0.230 | 21.4 | 0.235 | 21.9 | 0.033 | 3.1 | 0.245 | 22.8 |
| GRP5 | 270 | 3.57 | 0.108 | 3.0 | 0.040 | 1.1 | 0.049 | 1.4 | 0.575 | 16.1 | 0.588 | 16.5 | 0.082 | 2.3 | 0.615 | 17.3 |
| GRP6 | 270 | 7.53 | 0.132 | 1.8 | 0.031 | 0.4 | 0.055 | 0.7 | 0.445 | 5.9 | 0.468 | 6.2 | 0.000 | 0.0 | 0.509 | 6.8 |

Table 3. Reproducibility of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test evaluated on VITROS 3600 Immunodiagnostic Systems

a Between lot: Variability of the test performance from lot to lot.

b Within-Laboratory variability contains the Within Run, Between Run, Between Day and Between Lot variance components.

Between site: Variability of the test performance from site to site.

d Reproducibility: Variability of the test incorporating factors of site. lot, run and day.

+ %CVs are not meaningful for S/C results < 0.50

#### Table 4. Reproducibility of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test evaluated on VITROS 5600 Integrated Systems

| PanelMember | Number<br>of<br>Observ. | Grand<br>Mean<br>(S/C) | Repeatability<br>(Within Run) | | Between Run | | Between Day | | Between Lota | | Within-<br>Laboratoryb | | Between-Sitec | | Reproducibility<br>(Overall)d | |
|-------------|-------------------------|------------------------|-------------------------------|-----------|-------------|-----------|-------------|-----------|--------------|-----------|------------------------|-----------|---------------|-----------|-------------------------------|-----------|
| | | | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) | SD | CV<br>(%) |
| GRP1 | 270 | 0.00 | 0.000 | N/A+ | 0.000 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.002 | N/A+ | 0.002 | N/A+ |
| GRP2 | 270 | 1.05 | 0.085 | 8.1 | 0.000 | 0.0 | 0.000 | 0.0 | 0.045 | 4.3 | 0.097 | 9.2 | 0.000 | 0.0 | 0.119 | 11.3 |
| GRP3 | 270 | 0.29 | 0.015 | N/A+ | 0.002 | N/A+ | 0.004 | N/A+ | 0.020 | N/A+ | 0.025 | N/A+ | 0.002 | N/A+ | 0.031 | N/A+ |
| GRP4 | 270 | 1.08 | 0.050 | 4.6 | 0.009 | 0.8 | 0.027 | 2.5 | 0.204 | 18.9 | 0.212 | 19.7 | 0.000 | 0.0 | 0.221 | 20.5 |
| GRP5 | 270 | 3.58 | 0.103 | 2.9 | 0.017 | 0.5 | 0.099 | 2.8 | 0.483 | 13.5 | 0.504 | 14.1 | 0.000 | 0.0 | 0.538 | 15.1 |
| GRP6 | 270 | 7.55 | 0.137 | 1.8 | 0.006 | 0.1 | 0.161 | 2.1 | 0.251 | 3.3 | 0.328 | 4.3 | 0.000 | 0.0 | 0.484 | 6.4 |

al Between lot: Variability of the test performance from lot to lot.

b Within-Laboratory variability contains the Within Run, Between Run, Between Day and Between Lot variance components.

· Between site: Variability of the test performance from site to site.

d Reproducibility: Variability of the test incorporating factors of site, lot, run and day.

+ % CVs are not meaningful for S/C results < 0.50.

| Table 5. Reproducibility of VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG |
|----------------------------------------------------------------------------------|
| test evaluated on VITROS XT 7600 Integrated Systems |

| PanelMember | Number of Observ. | Grand Mean (S/C) | Repeatability (Within Run) | | Between Run | | Between Day | | Between Lota | | Within-Laboratoryb | | Between-Sitec | | Reproducibility (Overall)d | |
|-------------|-------------------|------------------|----------------------------|--------|-------------|--------|-------------|--------|--------------|--------|--------------------|--------|---------------|--------|----------------------------|--------|
| mber | Observ. | Mean (S/C) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| GRP1 | 270 | 0.00 | 0.001 | N/A+ | 0.000 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.001 | N/A+ | 0.002 | N/A+ |
| GRP2 | 270 | 1.06 | 0.035 | 3.4 | 0.000 | 0.0 | 0.010 | 0.9 | 0.039 | 3.7 | 0.053 | 5.1 | 0.009 | 0.9 | 0.061 | 5.7 |
| GRP3 | 270 | 0.29 | 0.012 | N/A+ | 0.000 | N/A+ | 0.007 | N/A+ | 0.023 | N/A+ | 0.027 | N/A+ | 0.003 | N/A+ | 0.029 | N/A+ |
| GRP4 | 270 | 1.09 | 0.037 | 3.4 | 0.000 | 0.0 | 0.013 | 1.2 | 0.220 | 20.2 | 0.224 | 20.6 | 0.013 | 1.1 | 0.229 | 21.1 |
| GRP5 | 270 | 3.59 | 0.097 | 2.7 | 0.000 | 0.0 | 0.000 | 0.0 | 0.510 | 14.2 | 0.519 | 14.5 | 0.018 | 0.5 | 0.537 | 15.0 |
| GRP6 | 270 | 7.54 | 0.118 | 1.6 | 0.000 | 0.0 | 0.032 | 0.4 | 0.380 | 5.0 | 0.399 | 5.3 | 0.000 | 0.0 | 0.457 | 6.1 |

a Between lot: Variability of the test performance from lot to lot

b Within-Laboratory variability contains the Within Run, Between Run, Between Lot variance components.

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

Between site: Variability of the test performance from site to site.

d Reproducibility: Variability of the test incorporating factors of site, lot, run and day.

+ % CVs are not meaningful for S/C results < 0.50.

- 2. Linearity:
Not applicable

- 3. Analytical Specificity:
- A. Cross-Reactivity: Cross-reactivity of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test was evaluated by testing serum samples with antibodies to other microorganisms or underlying conditions which could cause false positive results. Cross-reactivity was evaluated in the VITROS 5600 Integrated system, with samples tested in singlicate. No cross-reactivity was observed with any of the cross-reactants evaluated. The results for cross-reactivity are presented in Table 6 below.

| Sample Category | Number of<br>Samples Tested | VITROS Immunodiagnostic<br>Products<br>Anti-SARS-CoV-2 IgG test<br>Results | |
|------------------------------------------------|-----------------------------|----------------------------------------------------------------------------|----------|
| | | Non-reactive | Reactive |
| Influenza A Antibody | 10 | 10 | 0 |
| Influenza B Antibody | 12 | 12 | 0 |
| Hepatitis C Virus (HCV) Antibody | 15 | 15 | 0 |
| Hepatitis B Virus (HBV) Antibody | 12 | 12 | 0 |
| Haemophilus influenzae Antibody | 12 | 12 | 0 |
| Anti-Nuclear Antibody (ANA) | 10 | 10 | 0 |
| Rheumatoid Factor | 11 | 11 | 0 |
| Human Anti-Mouse Antibody (HAMA) | 10 | 10 | 0 |
| Adenovirus Antibody | 12 | 12 | 0 |
| Coxsackie B Virus Antibody | 10 | 10 | 0 |
| Echovirus Antibody | 10 | 10 | 0 |
| Poliovirus Antibody | 9 | 9 | 0 |
| Respiratory Syncytial Virus (RSV)<br>Antibody | 10 | 10 | 0 |
| Anti-SARS-coronavirus | 13 | 13 | 0 |
| Anti-MERS-coronavirus | 14 | 14 | 0 |
| Human coronavirus HKU1 Antibody | 11 | 11 | 0 |
| Human coronavirus NL63 Antibody | 10 | 10 | 0 |
| Human coronavirus OC43 Antibody | 21 | 21 | 0 |
| Human coronavirus 229E Antibody | 24 | 24 | 0 |
| Human Immunodeficiency Virus (HIV)<br>Antibody | 17 | 17 | 0 |
| Human Parainfluenza Virus (HPIV)<br>Antibody | 5 | 5 | 0 |
| Human Metapneumovirus (HMPV)<br>Antibody | 10 | 10 | 0 |

#### Table 6. Cross-reactivity study samples and results

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

| Sample Category | Number of<br>Samples Tested | VITROS Immunodiagnostic<br>Products<br>Anti-SARS-CoV-2 IgG test<br>Results | |
|--------------------------------------------------------|-----------------------------|----------------------------------------------------------------------------|----------|
| | | Non-reactive | Reactive |
| Enterovirus Antibody | 10 | 10 | 0 |
| Rhinovirus Antibody | 11 | 11 | 0 |
| Epstein-Barr Virus (EBV) Antibody | 11 | 11 | 0 |
| Epstein-Barr Virus Nuclear Antigen<br>(EBVNA) Antibody | 11 | 11 | 0 |
| Rubella Virus Antibody | 10 | 10 | 0 |
| Legionella pneumophila Antibody | 10 | 10 | 0 |
| Bordetella pertussis Antibody | 15 | 15 | 0 |
| Mycoplasma pneumoniae Antibody | 14 | 14 | 0 |
| Chlamydophila pneumoniae Antibody | 10 | 10 | 0 |
| Streptococcus pneumoniae Antibody | 10 | 10 | 0 |
| Streptococcus pyogenes Antibody | 10 | 10 | 0 |
| Candida albincans Antibody | 14 | 14 | 0 |
| Pneumocystis jirovecii Antibody | 10 | 10 | 0 |
| Pseudomonas aeruginosa Antibody | 14 | 14 | 0 |
| Staphylococcus epidermidis | 10 | 10 | 0 |
| Cytomegalovirus (CMV) Antibody | 11 | 11 | 0 |

- B. Interference: The VITROS Immunodiagnostic Products Anti SARS-CoV-2 IgG test was evaluated for interference consistent with CLSI document EP07. Commonly encountered substances were tested on one lot of reagent using the VITROS 5600 Integrated System in 3 reactive (low positive) and 3 non-reactive (negative) samples.
(0) +

To evaluate interference, the percent bias was calculated as:

All substances evaluated have been shown to not interfere (<10% bias for low reactive samples and bias <0.22 S/C for non-reactive samples) with the test performance at the concentration listed in the table below.

Table 7. Endogenous interferants evaluated with VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test

| | Test Concentration | | | | | |
|-------------------------|--------------------|-----------------|--|--|--|--|
| Substance | Conventional | SI Units<br>N/A | | | | |
| Anti-Nuclear Antibody | > 8 AI | | | | | |
| Bilirubin, conjugated | 40 mg/dL | 475 umol/L | | | | |
| Bilirubin, unconjugated | 40 mg/dL | 684 umol/L | | | | |
| Cholesterol | 400 mg/dL | 10.3 mmol/L | | | | |
| Hemoglobin | 1000 mg/dL | 10 g/L | | | | |
| Human anti-Mouse | 3600 ng/mL | 0.024 umol/L | | | | |
| Ig (total) | 6 g/dL | 60 g/L | | | | |

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

| | Test Concentration | |
|-------------------|--------------------|--------------|
| Substance | Conventional | SI Units |
| Rheumatoid Factor | 35.7-61.7 IU/mL | N/A |
| Total Protein | 15 g/dL | 150 g/L |
| Triglycerides | 1500 mg/dL | 16.94 mmol/L |

N/A: Not Applicable (alternative units are not provided)

Table 8. Exogenous interferants evaluated with VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test

| Substance | Test Concentration | |
|------------------|--------------------|---------------|
| | Conventional | SI Units |
| Abacavir | 1.27 mg/dL | 44.4 µmol/L |
| Acetaminophen | 15.6 mg/dL | 1.03 mmol/L |
| Amoxicillin | 1.35 mg/dL | 37 µmol/L |
| Aspirin | 3 mg/dL | 0.167 mmol/L |
| Atorvastatin | 75 µg/dL | 1.34 µmol/L |
| Azithromycin | 1.1 mg/dL | 14.8 µmol/L |
| Biotin | 3510 ng/mL | 14.3 µmol/L |
| Cefoxitin | 660 mg/dL | 15.5 mmol/L |
| Ceftriaxone | 84 mg/dL | 1.51 mmol/L |
| Dextromethorphan | 1.56 µg/dL | 0.0575 µmol/L |
| EDTA | 0.099 mg/dL | 3.39 µmol/L |
| Gentamicin | 3.0 mg/dL | 62.8 µmol/L |
| Guaifenesin | 450 µg/dL | 22.7 µmol/L |
| Heparin | 330 units/dL | 330 units/dL |
| Ibuprofen | 21.9 mg/dL | 1.06 mmol/L |
| Intralipid | 2000 mg/dL | N/A |
| Levofloxacin | 3.6 mg/dL | 99.7 µmol/L |
| Levothyroxine | 429 µg/dL | 0.552 µmol/L |
| Lisinopril | 24.6 µg/dL | 0.607 µmol/L |
| Lopinavir | 57.17 µg/mL | 90.89 µmol/L |
| Loratadine | 8.7 µg/dL | 0.271 µmol/L |
| Losartan | 1155 ng/mL | 2.505 µmol/L |
| Meropenem | 33.9 mg/dL | 884 µmol/L |
| Metformin | 1.2 mg/dL | 92.9 µmol/L |
| Metoprolol | 150 µg/dL | 5.61 µmol/L |
| Naproxen | 36.0 mg/dL | 1.56 mmol/L |
| Omeprazole | 840 µg/dL | 24.3 µmol/L |
| Oseltamivir | 39.9 µg/dL | 1.28 µmol/L |
| Peramivir | 183600 ng/mL | 559 µmol/L |
| Prednisone | 10 µg/dL | 0.276 µmol/L |
| Ritonavir | 10.98 mg/dL | 126.42 mmol/L |
| Theophylline | 6.0 mg/dL | 333 µmol/L |
| Vancomycin | 12.0 mg/dL | 82.8 µmol/L |
| Zanamivir | 1089 ng/mL | 3.28 µmol/L |

N/A = Not Applicable (alternative units are not provided)

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

When amlodipine was tested at a concentration of 1.88 ug/dL, a negative bias (-11.1% change of the S/C value) was observed in reactive samples.

- 4. Assay Reportable Range:
Not applicable

- 5. Traceability. Stability. Expected Values (Controls, Calibrators, or Methods):
Specimen Stability: The stability of SARS-CoV-2 antibodies in serum. K2-EDTA plasma. and K -- EDTA plasma was evaluated with the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test after various storage conditions with and without freeze-thawing cycles in the VITROS 5600 Integrated Systems. The storage conditions evaluated were room temperature, 2 - 8°C, and <- 20°C with 5 freeze-thaw cycles.

Freshly drawn whole blood from individual donors was collected. Of the whole blood donors, (1) were unaltered and were spiked using a 15/41

The whole blood was distributed among the collection tubes used in the study. Each spiked sample was unique.

After centrifugation, each sample from each collection tube was tested in duplicate using one reagent lot on one VITROS 5600 Integrated System, denoted as the fresh timepoint. Aliquots of each sample were prepared and stored at room temperature, 2-8°C. and <- 20°C. For the < 20℃ samples were prepared. 1 subjected to 5 freeze-thaw cycles (F/T) and the other subjected to (0) F/F/T cycles.

For each sample type and storage temperature the percent difference to baseline was calculated as follows where baseline is considered as the fresh sample before storing at any temperature condition:

(D (4)

The results support the following specimen storage conditions for serum, K-- EDTA and K --EDTA plasma:

| Sample Type | Temperature | Stability | F/T |
|-----------------------------------------------|-------------------------------|------------|-----|
| Serum and plasma<br>(K2-EDTA and K3-<br>EDTA) | Room Temperature<br>(15-30°C) | ≤ 24 hours | N/A |
| | Refrigerated<br>(2-8°C) | ≤ 7 days | N/A |
| | Frozen<br>(≤-20°C) | ≤ 4 weeks | N/A |

Table 9. Summary of specimen stability

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

| | N/A | N/A | 1 cycle |
|------------------------------------------------------|-----|-----|---------|
| A THE CHART COLLECT CHE CASE CONSTITUTION CONSULTION | | | |

N/A= Not Applicable

Based upon this study design and the results thereof, the specimen stability data support, storage of all matrices at: 15-30 ℃ for 24 hours, 2-8℃ for up to 7 days, and ≤-20℃ or below for up to five freeze-thaw cycle

- 6. Detection Limit:
Not applicable

- 7. Assay Cut-Off:
The study was performed to determine the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test S/C cutoff. The study included the testing of a collection of [0] megative samples collected prior to the COVID-19 pandemic, and samples collected from individuals with a prior SARS-CoV-2 RT-PCR positive result. Additionally, promogative samples collected prior to the pandemic were tested with reagent lots. All samples were tested in singlicate using one VITROS Anti-SARS-CoV-2 IgG reagent pack and calibrator lot. A Receiver Operating Characteristic (ROC) curve analyses was performed to optimize for those cutoff values that maximize both sensitivity specificity. At the cutoff S/C = 1.00 the resultant ROC was [b] confirming high sensitivity and specificity of the established S/C cutoff value (Figure 2).

Image /page/12/Figure/7 description: The image shows a figure labeled as "Figure 2:". There is also the text "(D)(4)" in the upper right corner of the figure. The image appears to be a gray rectangle with a red border. The text "Analytical sensitivity at the cutoff using a Certified Reference Material" is at the bottom of the image.

Analytical sensitivity at the cutoff using a Certified Reference Material

The analytical sensitivity of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test was determined using a series of serial dilutions of the WHO First International Standard for Anti-SARS-CoV-2 Immunoglobulin (human) code 20/136 (Certified Reference

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

Material or CRM) in negative patient sodium citrate plasma. Samples included in the study were prepared covering a will BAU/mL to | BAU/mL range (Table 10). Each sample was tested in duplicate using of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test on two VITROS analyzers (VITROS ECi/ECiQ and VITROS 3600 Immunodiagnostic Systems). In addition, an internal reference calibrator was also included in the assay.

Ortho-Clinical Diagnostics collected both ALU and S/C for each of the CRM samples tested. Ortho-Clinical Diagnostics analyzed the data using the least-squares regression analysis, where the [14] represented the BAU/mL and the [10] the S/C values for each sample. Using the regression equation, Ortho-Clinical Diagnostics calculated the BAU/mL at the cutoff (S/C = 1.00). The data analysis shows that a S/C of 1.00 corresponds to 100 |BAU/mL using the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test.

Table 10: CRM serial dilutions S/C results and calculation of BAU/mL at the cutoff

| CRM<br>concentrations<br>(BAU/mL) | VITROS ECi/ECiQ<br>(S/C) | VITROS 3600<br>(S/C) | VITROS System<br>Average S/C |
|-----------------------------------|--------------------------|----------------------|------------------------------|
| | (0)(4) | | |

Figure 3: Analytical sensitivity regression analysis.

Image /page/13/Picture/5 description: The image is a gray rectangle with a thin red border. In the top center of the rectangle is the text "(b)(4)". The rectangle takes up most of the image. The image is simple and contains no other objects.

{14}------------------------------------------------

# B Comparison Studies:

- 8. Method Comparison:
Not Applicable

- 9. Matrix Comparison:
A matrix equivalency study was conducted to support equivalent performance of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 1gG test between serum. K3-EDTA plasma, and K3-EDTA plasma. The study was conducted using 1 lot for the reagent pack, calibrator and controls in the VITROS 5600 Integrated Systems.

For this study unique paired clinical samples were evaluated (spiked samples prepared using a high titer convalescent plasma pool and non-spiked samples) with SARS-CoV-2 IgG antibodies levels representing the whole assay range (from low negative to high positive, including a 1046 of samples near the cutoff). After centrifugation, the matched serum, Ko-EDTA, and K3-EDTA samples were tested in duplicate using one reagent lot on 100 VITROS 5600 Integrated System. Sample distribution is summarized in Table 11 below.

Table 11. Sample distribution: negative, near the cutoff and positive

| Group | S/C Result | Number of<br>specimens (% total) |
|-------------------|------------|----------------------------------|
| Negative | | (b)(4) |
| Near cutoff | | |
| Moderate Positive | | |
| High Positive | | |
| Total | | (b)(4) |

A weighted Deming regression analysis comparing K2-EDTA plasma (the comparator matrix) to K2-EDTA plasma and to serum was conducted. The Deming regression analysis did not demonstrate significant deviation from the comparator matrix (Figure 4 - 5). Therefore, the study demonstrated equivalency between serum. K->EDTA plasma, and K-EDTA plasma.

Figure 4: Weighted Deming linear regression analysis between K2-EDTA plasma and serum

{15}------------------------------------------------

# C Clinical Studies:

- 10. Clinical Sensitivity:
Not applicable

# 11. Clinical Specificity:

Not applicable

# 12. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):

{16}------------------------------------------------

### Clinical Agreement Study:

The clinical performance of VITROS Immunodiagnostic Products anti SARS- CoV-2 IgG test was evaluated at three testing sites using 642 unique retrospective clinical samples acquired from two populations. Population 1 consisted of 338 samples collected from individuals previously infected with SARS-CoV-2 with a prior SARS-CoV-2 positive test result using a comparator that FDA determined is appropriate (RT PCR test). Population 2 consisted of 304 samples collected prior to December 2019 (before the widespread outbreak of COVID-19). Of the 304 samples, 30% of the tested samples were collected from blood donor centers.

Of the 338 samples in Population 1, 24 samples were collected 0 - 7 days from COVID-19 symptom onset, 21 samples collected between 8 - 14 days from symptom onset, and 293 samples were collected >15 days from COVID-19 symptom onset. Table 12 below shows the sample distribution and the respective percentages from the total of samples tested per "Days post-symptoms onset" time bin.

| Time bin<br>(n= total number<br>samples tested) | Days post-<br>symptom<br>onset | Percent Samples<br>(per time bin) |
|-------------------------------------------------|--------------------------------|-----------------------------------|
| 0-7 days (n = 24) | 0 - 4 | 70.83% |
| | 5 - 7 | 29.17% |
| 8-14 days (n = 21) | 8 - 11 | 47.62% |
| | 12 - 14 | 52.38% |
| ≥ 15 days (n = 293) | 15 - 21 | 10.58% |
| | 22 - 30 | 10.92% |
| | 31 - 60 | 40.61% |
| | 61 - 90 | 26.62% |
| | 91 - 195 | 11.26% |

Table 12. Sample distribution within each "Days post symptoms onset" time bin.

Tables 13 and 14 below represents sample distribution per matrix tested for each study population (Population 1 and Population 2).

| Table 13. Distribution of Population I samples by matrix and days post-symptom onset. | | | | |
|---------------------------------------------------------------------------------------|-------------------------|-------------|----------|-------|
| Matrix | Days post-symptom onset | | | |
| | ≤7 days | 8 - 10 days | ≥15 days | Total |
| EDTA plasma | | (b)(4) | | 169 |
| Serum | | | | 169 |
| Total | | | | 338 |

Table 14. Distribution of Population 2 samples by matrix.

{17}------------------------------------------------

| EDTA plasma | 138 |
|-------------|-----|
| Serum | 166 |
| Total | 304 |

Ortho conducted the clinical agreement study at 3 testing sites: one internal site and 2 external sites.

# Positive Percent Agreement (PPA)

A total of 338 samples collected from individual patients confirmed to have a prior SARS-CoV-2 positive result by RT-PCR were tested. Blood samples were collected within the United States between April 2020 and March 2021. Samples were tested with the VITROS Immunodiagnostic Products Anti SARS-CoV-2 IgG test in each analyzer (except with the VITROS XT7600 Integrated System where one sample was not tested due to limited volume). Of the 338 samples included in the study, 169 were EDTA plasma samples and 169 were serum samples.

The performance of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test and 95% Confidence Interval for each VITROS analyzer is summarized in the tables below.

PPA performance of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test and 95% Confidence Interval by system:

| Days from<br>Symptom<br>Onset | Number of<br>Subjects Tested<br>(with prior RT-<br>PCR Positive) | Reactive | PPA | 95% CI<br>(Wilson score) |
|-------------------------------|------------------------------------------------------------------|----------|--------|--------------------------|
| 0-7 days | 24 | 11 | 45.83% | 27.89%-64.93% |
| 8-14 days | 21 | 11 | 52.38% | 32.37% - 71.66% |
| ≥15 days | 293 | 275 | 93.86% | 90.50%-96.10% |
| Total | 338 | -- | -- | -- |

Table 15. In the VITROS ECi/ECiO Immunodiagnostic Systems.

Table 16. In the VITROS 3600 Immunodiagnostic Systems and the VITROS 5600 Integrated Systems

| | | VITROS Immunodiagnostic Products<br>Anti-SARS-CoV-2 IgG test Results | | |
|----------------------------|------------------------------------------------------------------|----------------------------------------------------------------------|--------|--------------------------|
| Days from<br>Symptom Onset | Number of<br>Subjects Tested<br>(with prior RT-<br>PCR Positive) | Reactive | PPA | 95% CI<br>(Wilson score) |
| 0-7 days | 24 | 10 | 41.67% | 24.47% - 61.17% |
| 8-14 days | 21 | 11 | 52.38% | 32.37% - 71.66% |
| ≥15 days | 293 | 274 | 93.52% | 90.10% - 95.81% |
| Total | 338 | -- | -- | -- |

{18}------------------------------------------------

| Days from<br>Symptom<br>Onset | Number of<br>Subjects Tested<br>(with prior RT-<br>PCR Positive) | Reactive | PPA | 95% CI<br>(Wilson score) |
|-------------------------------|------------------------------------------------------------------|----------|--------|--------------------------|
| 0-7 days | 24 | 10 | 41.67% | 24.47% - 61.17% |
| 8-14 days | 21 | 11 | 52.38% | 32.37% - 71.66% |
| ≥15 days | 292 | 273 | 93.49% | 90.10% - 95.80% |
| Total | 337 | -- | -- | -- |

Table 17. In the VITROS XT 7600 Integrated Systems

# Negative Percent Agreement (NPA)

Three hundred and four (304) presumed SARS-CoV-2 negative samples collected prior to the COVID-19 pandemic within the United States were tested. Of the 304 samples, 138 were EDTA plasma samples and 166 serum samples. All samples were tested using VITROS ECi/ECiQ/3600 Immunodiagnostic Systems and the VITROS 5600/XT 7600 Integrated Systems "(except the samples that were not tested on one or more analyzers due to limited volume). The performance of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test and 95% Confidence Interval for each VITROS analyzer is summarized in the table below:

Table 18: NPA performance of the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test and 95% confidence interval in all VITROS analyzers

| | | VITROS Immunodiagnostic Products<br>Anti-SARS-CoV-2 IgG test Results | | |
|--------------------------------------------|--------------------------------------------|----------------------------------------------------------------------|--------|--------------------------|
| Analyzer | Presumed Negative<br>(Collected Pre-COVID) | Non-<br>Reactive | NPA | 95% CI<br>(Wilson score) |
| VITROS ECi/ECiQ/VITROS<br>3600/VITROS 5600 | 304 | 301 | 99.01% | 97.14%-<br>99.66% |
| VITROS XT 7600 | 303 | 300 | 99.01% | 97.13% -<br>99.66% |

# D Clinical Cut-Off:

Not applicable

# E Expected Values/Reference Range:

Not applicable

### F Other Supportive Performance Characteristics Data:

{19}------------------------------------------------

# Calibration Cvcle Stability

The purpose of this study was to establish the calibration interval, or how frequently the assay should be calibrated. The calibration interval was established by testing samples on the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test using a single calibration over time.

In the study, Ortho-Clinical Diagnostics included a panel of 6 precision pool samples. Each panel member was prepared as follows:

- Precision Pool 1 (PP1): Control level 1 (Non-Reactive) .
- . Precision Pool 2 (PP2): Control level 2 (Reactive)
- Precision Pool 3 (PP3): Calibrator .
- Precision Pool 4 (PP4): High negative EDTA plasma sample .
- . Precision Pool 5 (PP5): Low Positive EDTA plasma sample
- Precision Pool 6 (PP6): High Positive EDTA plasma sample .

After a single initial calibration on Day samples were tested on Day and up to Day on the VITROS ECi/ECiO, VITROS 3600, VITROS 5600, and VITROS XT7600.

For each positive sample (PP2, PP4, PP5, and PP6) linear regression analysis was conducted. In addition, percent difference from baseline was calculated as follows:

| (b)(4) |
|--------|
|--------|

Percent difference to baseline should be (b)(4).The percent difference to baseline should be

The study results support a calibration interval stability of 28 days for the VITROS Immunodiagnostic Products Anti-SARS-CoV-2 IgG test.

#### VII Proposed Labeling:

The labeling supports the decision to grant the De Novo request for this device.

#### VIII Identified Risks and Mitigations:

| Risks to Health | Mitigation Measures |
|----------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Risk of false test results | Certain labeling information including<br>limitations, device descriptions, explanations<br>of procedures and performance information<br>identified in special controls (1), (3), and (5).<br>Use of certain specimen collection devices<br>identified in special control (2).<br>Certain design verification and validation<br>including documentation of device<br>descriptions, certain analytical studies and<br>clinical studies, and risk analysis strategies<br>identified in special control (4).<br>Testing of characterized samples and labeling<br>information identified in special control (6). |

10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov DEN210038 -

{20}------------------------------------------------

| Risks to Health | Mitigation Measures |
|-------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Failure to correctly interpret the test results | Certain labeling information including<br>limitations, device descriptions, explanations<br>of procedures and performance information<br>identified in special controls (1), (3), and (5).<br>Use of certain specimen collection devices<br>identified in special control (2).<br>Certain design verification and validation<br>including documentation of device<br>descriptions, certain analytical studies and<br>clinical studies, and risk analysis strategies<br>identified in special control (4).<br>Testing of characterized samples and labeling<br>information identified in special control (6). |
| Failure to correctly operate the device | Certain labeling information including<br>limitations, device descriptions, explanations<br>of procedures and performance information<br>identified in special controls (1), (3), and (5).<br>Use of certain specimen collection devices<br>identified in special control (2). |

#### IX Benefit/Risk Assessment:

### A Summary of the Assessment of Benefit:

The benefit of the assay is the ability to detect Anti-SARS-CoV-2 IgG antibodies as an aid in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection. The device could also provide a tool for tracking possible patient exposure.

True positive test results provide additional support for the diagnosis of recent or past SARS-CoV-2 infection. Results could be used in conjunction with clinical and epidemiological information, as well as other laboratory results to guide patient management. The test results may improve infection control measures and may aid in tracking and reducing transmission of infection. There is currently no SARS-CoV-2 antibody test that has undergone full FDA premarket review to most definitively determine clinical truth of the presence or absence of detectable antibodies for the method comparison study, however this uncertainty could be acceptable, particularly because the sponsor used a comparator that FDA determined is appropriate (SARS-CoV-2 RT-PCR devices), which represents the most reasonable alternative to establish clinical truth in the clinical study. This is an acceptable source of uncertainty regarding the benefits of the test.

#### Summary of the Assessment of Risk: B

The risks associated with the device, when used as intended, are those related to the risk of false test results, which have essentially the same impacts as the risks related to failure to correctly

{21}------------------------------------------------

interpret the test results and failure to correctly operate the device as all would cause the user to rely on incorrect information.

A false negative result could be interpreted as indicating that a person did not recently have COVID-19, which may lead a person to take fewer necessary precautions against spreading the virus to others if they are still shedding the virus from a recent infection. This may increase the risk of transmission. In the context of the current public health emergency, incorrect serological test results used to guide infection control activities could lead to misallocation of resources used for surveillance and prevention. The positive percent agreement performance point estimate of the device observed in the clinical study indicates that false negative results are not likely to occur when the device is used in the intended use population.

A false positive SARS-CoV-2 antibody result could be interpreted as a diagnosis of recent COVID-19, and a clinician may assume a patient may still be shedding the virus, which may result in unnecessary additional testing, quarantine, or self-isolation to prevent the spread of the virus to others. False positive serology test results can lead to an incorrect assessment that the tested person had an immune response to SARS-CoV-2, which may lead the person to take fewer necessary precautions against virus exposure. This may increase the individual's risk of infection and may lead the person to not seek testing if later infected, likely increasing the spread of the disease. In the context of the current public health emergency, incorrect serological test results used to guide infection control activities could lead to misallocation of resources used for surveillance and prevention.

A positive result could be wrongly interpreted as a diagnosis of acute COVID-19 to explain an individual's symptoms and delay correct diagnosis and initiation of appropriate treatment for the actual cause of patient illness. A positive test result could be wrongly interpreted as indicating that the tested person has immunity to SARS-CoV-2, which may lead the person to take fewer precautions against virus exposure. This may increase the individual's risk of infection and may lead the person to not seek testing if later infected, likely increasing the spread of the disease. A negative result may be misinterpreted as ruling out SARS-CoV-2 infection, with a concomitant delay in the correct diagnosis and treatment.

# C Patient Perspectives:

This submission did not include specific information on patient perspectives for this device.

# D Summary of the Assessment of Benefit-Risk:

The risks associated with the device (risk of false test results, failure to correctly interpret the results, and failure to correctly operate the device) are mitigated by labeling information, which will assist the operator in correctly performing the test and will assist healthcare providers in understanding the intended use of the test and evaluating the predictive value of a result based on the analytical and clinical performance of the test. In addition, those risks are mitigated by the use of certain validated specimen collection devices. Further, the risk of false test results and failure to correctly interpret the results are mitigated by certain design verification and validation, including analytical and clinical studies and risk analysis strategies to reduce the likelihood of such errors. Finally, the risk of false test results due to a disease or disorder that Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov DEN210038 -

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presents a public health emergency, or a public health emergency that otherwise exists are addressed by special controls requiring certain testing of characterized samples and labeling information in those situations. The special controls help to ensure that errors will be uncommon and will facilitate accurate assay implementation of results. In addition, the device's performance observed in the clinical study suggests that errors will be uncommon and that the assay will provide benefits to patients as an aid in identifying individuals with an adaptive immune response to SARS-CoV-2, indicating recent or prior infection. While general controls alone are insufficient to mitigate the risks associated with the device, the benefits outweigh the risks given the special controls.

#### Conclusion: X

The De Novo request is granted, and the device is classified under the following regulation and subject to the special controls identified in the letter granting the De Novo request:

| Product Code(s): | QVP |
|------------------|--------------------------|
| Device Type: | SARS-CoV-2 serology test |
| Class: | Class II |
| Regulation: | 21 CFR 866.3983 |