Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on standard immunoassay technology (ECLIA) and statistical analysis of the resulting sFlt-1/PlGF ratio. There is no mention of AI, ML, or related concepts in the provided text.

Therapeutic

No
This device is an in vitro diagnostic immunoassay that aids in the risk assessment of preeclampsia, but it does not directly treat or prevent a disease or condition.

Diagnostic

Yes

The device quantitatively determines the sFlt-1/PlGF ratio in human serum, which is explicitly "indicated as an aid in the risk assessment of pregnant women... to develop preeclampsia with severe features within two weeks from testing," directly providing information to assess a medical condition.

SaMD (Software as a Medical Device)

No

The device is an immunoassay system that relies on physical reagents and a dedicated immunoassay analyzer (cobas e) to perform the test and generate results. While software is likely involved in controlling the analyzer and processing the data, the core functionality and components are hardware and chemical-based, not software-only.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states "Immunoassays for the in vitro quantitative determination..." and "The sFlt-1/PlGF ratio is indicated as an aid in the risk assessment of pregnant women...". The term "in vitro" is a key indicator of an IVD.
Sample Type: The device analyzes "human serum," which is a biological sample taken from the body, a common characteristic of IVDs.
Purpose: The purpose is to provide information ("aid in the risk assessment") about a medical condition (preeclampsia) by analyzing a sample outside of the body.
Technology: The description details an "electrochemiluminescence immunoassay 'ECLIA'", which is a laboratory-based analytical technique used for diagnostic purposes.
Clinical Performance Evaluation: The document describes a clinical study (PRAECIS study) to evaluate the performance of the device in a clinical setting, which is a requirement for IVDs.
Predicate Device: The mention of a "Predicate Device(s)" with a K number (DEN220027) indicates that this device is being compared to a previously cleared IVD by a regulatory body like the FDA.

All of these factors strongly indicate that this device is an In Vitro Diagnostic.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Immunoassays for the in vitro quantitative determination of the tyrosine kinase-1/placental growth factor (sFlt-1/PlGF) ratio in human serum.

The sFlt-1/PlGF ratio is indicated as an aid in the risk assessment of pregnant women, with a singleton pregnancy (23+0 to 34+6/7 weeks' gestation) hospitalized for hypertensive disorders of pregnancy (preeclampsia, chronic hypertension with or without superimposed preeclampsia, or gestational hypertension), to develop preeclampsia with severe features within two weeks from testing. The sFit-1/PIGF ratio should be used in conjunction with clinical assessment and routine laboratory testing.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

Product codes (comma separated list FDA assigned to the subject device)

QWH

Device Description

The Elecsys sFlt-1 and Elecsys PlGF assays employ a sandwich principle.
For Elecsys sFlt-1:
1st incubation: 20 µL of sample, a biotinylated monoclonal sFlt-1 specific antibody, and a monoclonal sFlt-1 specific antibody labeled with a ruthenium complex (Tris(2.2'bipyridyl)ruthenium(II)-complex (Ru(bpy)s') react to form a sandwich complex.
2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.
The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
Results are determined via a calibration curve which is instrument-specifically generated by 2point calibration and a master curve provided via the reagent barcode.
The Elecsys sFlt-1 reagents "M", "R1" and "R2" are combined in a "rackpack".

For Elecsys PlGF:
1st incubation: 50 µL of sample, a biotinylated monoclonal PlGF specific antibody and a monoclonal PlGF specific antibody labeled with a ruthenium complex (Tris(2.2'bipyridyl)ruthenium(II)-complex (Ru(bpy)) react to form a sandwich complex.
2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.
The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
Results are determined via a calibration curve which is instrument-specifically generated by 2point calibration and a master curve provided via the reagent barcode.
The Elecsys PlGF reagents "M", "RI" and "R2" are combined in a "rackpack".

Both assays contain a blocking agent for exogenous biotin from supplements.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Pregnant women (≥18 years of age)

Intended User / Care Setting

Not Found

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

Not Found

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

Not Found

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

Non-Clinical Performance Evaluation:

Precision:
- Study Type: Evaluated repeatability (within-run precision) and intermediate precision (within-laboratory precision) according to CLSI guideline EP05-A3.
- Sample Size: One reagent lot, 2 aliquots of each control (PC Multimarker) and human sera (HS) in single determinations per run, 2 runs per day for 21 days for a total of 84 determinations for most samples. For Human serum 4 (PlGF), n=83.
- Key Results (PIGF): CVs for repeatability ranged from 1.0% to 5.7%. CVs for intermediate precision ranged from 1.7% to 9.9%.
- Key Results (sFlt-1): CVs for repeatability ranged from 0.9% to 2.4%. CVs for intermediate precision ranged from 1.7% to 3.7%.
- Key Results (Ratio): CVs for repeatability ranged from 1.1% to 4.9%. CVs for intermediate precision ranged from 1.4% to 7.0%.
Linearity/Assay Reportable Range:
- Study Type: Linearity study conducted with Elecsys sFlt-1 and PlGF assays on the cobas e 411 analyzer, according to CLSI EP06-Ed2, Study Design B.
- Sample Size: More than two sets of panel members, diluted samples covering the entire measuring range. Assayed in 4-fold determination within a single run.
- Key Results: Reportable range for Elecsys sFlt-1 is 80-85000 pg/mL. Reportable range for Elecsys PlGF is 10-5400 pg/mL.
Analytical Sensitivity (LoB, LoD, LoQ):
- LoB (Limit of Blank): Determined according to CLSI EP17-A2. 60 determinations for analyte-free samples. LoB claim in labeling: PlGF ≤ 2 pg/mL, sFlt-1 26.4 mg/dL can result in up to 10% decrease of sFlt-1/PlGF ratio around the cutoff of 38. Heterophilic human anti-mouse antibodies (HAMA) and high titers of antibodies to analyte-specific antibodies, streptavidin or ruthenium can interfere.
Analytical Specificity/Cross-Reactivity:
- Study Type: Cross-reactivity study with human serum samples in the presence and absence of potential cross-reactants.
- sFlt-1 Key Results: VEGFR2 ( 500 U/L. No interference found with the additional 13 substances.

Clinical Performance Evaluation:

PRAECIS Study ("Preeclampsia Risk Assessment: Evaluation of Cut-offs to Improve Stratification"):
- Study Type: Validation of the Roche sFlt-1/PlGF ratio cut-off of 38.
- Sample Size: Total N = 556 pregnant women (≥18 years of age, gestational age 23+0 to 34+6/7 weeks) hospitalized for hypertensive disorders of pregnancy.
- Key Results (sFlt-1/PIGF ratio at cutoff of 38):
  - Sensitivity: 91.40% (95% CI: 86.41%, 95.00%)
  - Specificity: 77.30% (95% CI: 72.68%, 81.47%)
  - NPV: 94.70% (95% CI: 91.54%, 96.94%)
  - PPV: 66.93% (95% CI: 60.77%, 72.68%)
  - Accuracy: 82.01% (95% CI: 78.56%, 85.12%)
  - LR-: 0.11 (95% CI: 0.07, 0.18)
  - LR+: 4.03 (95% CI: 3.32, 4.88)
- Impact of hypertensive disorder at presentation:
  - Preeclampsia (n=142): Sensitivity 95.06%, Specificity 72.13%, PPV 81.91%, NPV 91.67%.
  - Superimposed Preeclampsia (n=68): Sensitivity 93.75%, Specificity 55.56%, PPV 65.22%, NPV 90.91%.
  - Chronic Hypertension (n=232): Sensitivity 87.76%, Specificity 82.51%, PPV 57.33%, NPV 96.18%.
  - Gestational Hypertension (n=114): Sensitivity 83.33%, Specificity 78.89%, PPV 51.28%, NPV 94.67%.
- Impact of maternal age at presentation:
  *

Regulation Number and Section

N/A

Summary

0

Image /page/0/Picture/0 description: The image shows the date February 7, 2025. The text is written in a simple, sans-serif font. The date is written out in full, with the month, day, and year clearly visible. The text is black and the background is white.

Image /page/0/Picture/1 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 consists of the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.

Roche Diagnostics Jane Phillips Senior Regulatory Manager 9115 Hague Road Indianapolis, Indiana 46250

Re: K241453

Trade/Device Name: Elecsys sFlt-1 and Elecsys PlGF Regulation Number: 21 CFR 862.1602 Regulation Name: Prognostic Test for Development or Progression of Preeclampsia Regulatory Class: Class II Product Code: QWH Dated: January 2, 2025 Received: January 6, 2025

Dear Jane Phillips:

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.

1

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).

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.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

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-regulatory

2

assistance/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, Joseph A. Kotarek -S Joseph Kotarek Branch Chief Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

3

Indications for Use

510(k) Number (if known) K241453

Device Name Elecsys sFlt-1 and Elecsys PlGR

Indications for Use (Describe)
--------------------------------	--

Immunoassays for the in vitro quantitative determination of the tyrosine kinase-1/placental growth factor (sFlt-1/PlGF) ratio in human serum.

The sFlt-1/PlGF ratio is indicated as an aid in the risk assessment of pregnant women, with a singleton pregnancy (23+0 to 34+6/7 weeks' gestation) hospitalized for hypertensive disorders of pregnancy (preeclampsia, chronic hypertension with or without superimposed preeclampsia, or gestational hypertension), to develop preeclampsia with severe features within two weeks from testing. The sFit-1/PIGF ratio should be used in conjunction with clinical assessment and routine laboratory testing.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

| Over-The-Counter Use (21 CFR 801 Subpart C)

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K241453 Elecsys sFlt-1 and Elecsys PIGF 510(k) Summary

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

Submitter Name	Roche Diagnostics
Address	9115 Hague Road
P.O. Box 50416
Indianapolis, IN 46250-0457
Contact	Dr. Jane Phillips
Phone: (317) 521-3338
Email: jane.phillips@roche.com
Date Prepared	February 2nd, 2025
Proprietary Name	Elecsys sFlt-1 and Elecsys PIGF
Common Name	Elecsys PE Assays
Classification Name	Prognostic test for development or progression of preeclampsia
Product Codes,
Regulation Numbers	QWH, 21CFR862.1602
Predicate Devices	B·R·A·H·M·S sFlt-1/ PIGF KRYPTOR Test System (DEN220027)
Establishment Registration	Roche Diagnostics GmbH Mannheim, Germany: 9610126
Roche Diagnostics GmbH Penzberg, Germany: 9610529
Roche Diagnostics Indianapolis, IN United States: 1823260

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DEVICE DESCRIPTION 1.

1.1. Elecsys sFlt-1

The Elecsys sFlt-1 employs a sandwich principle. The total duration of the assay is 18 minutes.

1st incubation: 20 µL of sample, a biotinylated monoclonal sFlt-1 specific antibody, and a monoclonal sFlt-1 specific antibody labeled with a ruthenium complex (Tris(2.2'bipyridyl)ruthenium(II)-complex (Ru(bpy)s') react to form a sandwich complex.

2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.

The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.

Results are determined via a calibration curve which is instrument-specifically generated by 2point calibration and a master curve provided via the reagent barcode.

The Elecsys sFlt-1 reagents "M", "R1" and "R2" are combined in the so-called "rackpack", a bundle of the three reagent bottles, which is placed on the instrument as a single unit. The reagent rackpack is labeled as SFLT-1.

M: Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL: . Streptavidin-coated microparticles 0.72 mg/mL; preservative.
R1: Anti-sFlt-1-Ab~biotin (gray cap), 1 bottle, 9 mL: Biotinylated monoclonal anti . sFlt-1 antibody (mouse) 0.5 mg/L; phosphate buffer 100 mmol/L, pH 7.2; preservative.
R2: Anti-sFlt-1-Ab~Ru(bpy) (black cap), 1 bottle, 9 mL: Monoclonal anti sFlt-1 . antibody (mouse) labeled with ruthenium complex 1.0 mg/L, biotin scavenger antibody 1.0 mg/mL; phosphate buffer 100 mmol/L, pH 7.2; preservative.

6

Elecsys sFlt-1 will be marketed with the following devices:

CalSet sFlt-1; secondary, calibrator; JIT, 862.1150 Class II, exempt ●
Elecsys PreciControl Multimarker (cleared in K102157); multi-analyte controls, all . kinds (assayed); JJY, 862.1660 – Class I, exempt

1.2. Elecsys PIGF

The Elecsys PIGF employs a sandwich principle. The total duration of the assay is 18 minutes.

1 st incubation: 50 µL of sample, a biotinylated monoclonal PIGF specific antibody and a monoclonal PlGF specific antibody labeled with a ruthenium complex (Tris(2.2'bipyridyl)ruthenium(II)-complex (Ru(bpy)) react to form a sandwich complex.

2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.

The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.

Results are determined via a calibration curve which is instrument-specifically generated by 2point calibration and a master curve provided via the reagent barcode.

The Elecsys PlGF reagents "M", "RI" and "R2" are combined in the so-called "rackpack", a bundle of the three reagent bottles, which is placed on the instrument as a single unit. The reagent rackpack is labeled as PLGF.

M Streptavidin-coated microparticles (transparent cap). 1 bottle. 6.5 mL: . Streptavidin-coated microparticles 0.72 mg/mL; preservative.
R 1 Anti-PlGF-Ab~biotin (gray cap), 1 bottle, 8 mL: Biotinylated monoclonal anti . PlGF antibody (mouse) 0.6 mg/L; phosphate buffer 50 mmol/L, pH 6.0; preservative.

7

. R2 Anti-PlGF-Ab~Ru(bpy) (black cap), 1 bottle, 8 mL: Monoclonal anti PlGF antibody (mouse) labeled with ruthenium complex 4.0 mg/L, biotin scavenger antibody 1.0 mg/mL; phosphate buffer 50 mmol/L, pH 6.0; preservative.
Elecsys PlGF will be marketed with the following devices:
CalSet PlGF; secondary, calibrator; JIT, 862.1150 Class II, exempt .
. PreciControl Multimarker (cleared in K102157); multi-analyte controls, all kinds (assayed); JJY, 862.1660 - Class I, exempt

Both assays contain a blocking agent for exogenous biotin from supplements.

INDICATIONS FOR USE 2.

Immunoassays for the in vitro quantitative determination of the soluble fms like tyrosine kinase-1/placental growth factor (sFlt-1/PlGF) ratio in human serum.

The sFlt-1/PlGF ratio is indicated as an aid in the risk assessment of pregnant women, with a singleton pregnancy (23+0 to 34+6/7 weeks' gestation) hospitalized for hypertensive disorders of pregnancy (preeclampsia, chronic hypertension with or without superimposed preeclampsia, or gestational hypertension), to develop preeclampsia with severe features within two weeks from testing. The sFit-1/PlGF ratio should be used in conjunction with clinical assessment and routine laboratory testing.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

3. IMMUNOASSAY ANALYZERS.TECHNOLOGICAL CHARACTERISTICS

The following tables compare the Elecsys sFlt-1 and PlGF with its predicate device, B·R·A·H·M·S sFlt-1/ PlGF KRYPTOR Test System (DEN220027).

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| Feature Elecsys sFlt-1 and PIGF B·R·A·H·M·S sFlt-1/ PIGF
KRYPTOR Test System
DEN220027 |
--------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Immunoassays for the in vitro
| The B·R·A·H·M·STM sFlt-1/
| | between gestational age 23+0
|
| Sandwich immunoassay | Same |
| Electrochemiluminescence
| Time-Resolved Amplified
|
| 18 minute | 9 minute |
| Human serum | Human serum/plasma |
| N/A | K2-EDTA |
| Liquid, ready to use | Same |
| Phosphate buffer | Bovine albumin |
| sFlt-1 CalSet and PIGF CalSet | B·R·A·H·M·S sFlt-1
|
| 2-point calibration | Same |
| Calibration must be performed

• After 28 days using the
same reagent lot

• After 7 days (when
using the same reagent
kit on the analytzer)

• As required: e.g.
quality control findings | Calibration must be carried out
before the first use of each new
B·R·A·H·M·S sFlt-1 and PIGF
KRYPTOR lot, then
repeated on a regular basis
automatically managed by the
B·R·A·H·M·S KRYPTOR
analyzer in order to readjust the
standard curve. Controls must
be run after each calibration. |
| | outside the defined
limits | |
| Controls | PreciControl Multimarker | B·R·A·H·M·S sFlt-1
KRYPTOR QC kit and
B·R·A·H·M·S PIGF
KRYPTOR QC kit |
| Traceability/Standardization | Elecsys sFlt-1 assay: This
method has been standardized
against the
Quantikine ELISA Human
VEGFR1/Flt-1 Immunoassay.
• Elecsys PIGF assay: This
method has been standardized
against the
Quantikine ELISA Human
PIGF Immunoassay. | An international PIGF
reference preparation is not
available. Therefore, the assay
is calibrated using a reference
preparation of recombinant
human PIGF as the "Highest
Available Standard". This
calibration is verified on a
regular basis by regression
analysis and comparison to
theoretically expected results.
The PIGF assay results are
given in pg/mL. |
| Reagent Stability | Reagent stability for on
analyzer is now 28 days. | unopened at 2-8 °C: up
to the stated expiration
date after opening at 2-8 °C:
12 weeks on analyzers: 8 weeks |
| Measuring Range | sFlt-1 80-85000 pg/mL
PIGF 10-5400 pg/mL | sFlt-1 315-90,000 pg/mL
PIGF 7.6 – 4,000 pg/mL |
| Biotin Tolerance | Up to 1200 ng/mL | N/A |

Table 1: Technical Characteristics Comparison Table between Elecsys sFlt-1 and PIGF and B·R·A·H·M·S sFlt-1/ PlGF KRYPTOR Test System

9

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NON-CLINICAL PERFORMANCE EVALUATION 4.

Non-clinical performance evaluation for Elecsys sFit-1 and Elecsys P1GF assay executed with the study briefly summarized.

4.1. Precision

Precision measurements were conducted to evaluate repeatability (within-run precision) and intermediate precision (within-laboratory precision) according the CLSI guideline EP05-A3. Precision of the Elecsys sFlt-1 and PlGF assays was evaluated on a single cobas e 411 immunoassay analyzer according to CLSI EP5-A3 guideline. One reagent lot was evaluated. The precision study was conducted using the study design of testing 2 aliquots of each control

11

(PC (=PreciControl) Multimarker) and human sera (HS) in single determinations per run, 2 runs per day for 21 days. Assay calibration was performed as specified in the package insert. The samples were run in randomized order on the cobas e 411 analyzer.

	Mean	Repeatability		Intermediate precision
Sample	pg/mL	SD
(pg/mL )	CV (%)	SD
(pg/mL )	CV (%)	n
Human serum 1	46.3	1.03	2.2	1.51	3.3	84
Human serum 2	89.9	1.18	1.3	2.03	2.3	84
Human serum 3	441	4.46	1.0	7.27	1.7	84
Human serum 4	11.8	0.670	5.7	1.17	9.9	83
Human serum 5	74.0	1.87	2.5	4.11	5.6	84
Human serum 6	39.4	1.06	2.7	2.09	5.3	84
Human serum 7	96.0	1.58	1.6	2.38	2.5	84
Human serum 8	74.7	1.34	1.8	2.28	3.1	84
Human serum 9	4923	78.4	1.6	116	2.4	84
Human serum 10	8894	141	1.6	178	2.0	84
PCa) Multimarker 1	96.9	1.47	1.5	2.26	2.3	84
PC Multimarker 2	950	8.68	0.9	17.1	1.8	84

Table 2: Summary of 21-Day Precision for Elecsys PlGF on the cobas e 411 analyzer

a)

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	Mean	Repeatability		Intermediate precision
Sample	pg/mL	SD
(pg/mL )	CV (%)	SD
(pg/mL )	CV (%)	n
Human serum 1	1690	21.1	1.2	28.2	1.7	84
Human serum 2	632	6.45	1.0	11.1	1.8	84
Human serum 3	3867	43.0	1.1	74.1	1.9	84
Human serum 4	89.3	2.02	2.3	3.15	3.5	83
Human serum 5	36992	682	1.8	1287	3.5	84
Human serum 6	76873	1870	2.4	2842	3.7	84
Human serum 7	18037	246	1.4	405	2.2	84
Human serum 8	2522	28.6	1.1	46.0	1.8	84
Human serum 9	78.6	1.85	2.3	2.77	3.5	84
Human serum 10	90.7	2.02	2.2	3.32	3.7	84
PCa) Multimarker 1	106	2.13	2.0	2.98	2.8	84
PC Multimarker 2	1044	9.46	0.9	17.3	1.7	84

Table 2: Summary of 21-Day Precision for Elecsys sFlt-1 on the cobas e 411 analyzer

Table 3: 21-Day Precision Results Ratio Summary

Sample	Mean	Repeatability		Intermediate precision
	Ratio	SD	CV (%)	SD	CV (%)	n
Human serum 1	37	0.736	2.0	0.985	2.7	84
Human serum 2	7	0.118	1.7	0.133	1.9	84
Human serum 3	9	0.094	1.1	0.123	1.4	84
Human serum 4	8	0.369	4.9	0.528	7.0	83
Human serum 5	501	17.9	3.6	27.4	5.5	84
Human serum 6	1957	71.8	3.7	101	5.1	84
Human serum 7	188	2.95	1.6	3.92	2.1	84
Human serum 8	34	0.497	1.5	0.798	2.4	84
Human serum 9	0.016	0.0005	2.9	0.0005	3.4	84
Human serum 10	0.01	0.0003	2.5	0.0004	3.7	84
PC Multimarker 1	1	0.025	2.3	0.026	2.3	84
PC Multimarker 2	1	0.015	1.4	0.017	1.5	84

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4.2. Linearity/Assay Reportable Range

The linearity study was conducted with the Elecsys sFlt-1 and PIGF assays on the cobas e 411 analyzer to demonstrate that measurements across the claimed measuring range for each parameter are linear. This was done according to CLSI EP06-Ed2. Study Design B. The linearity study was performed with more than two sets of panel members with each set partially covering the proposed linearity interval. The main reason for this study design is the large measuring range and limitations in the number of dilutions of each high sample. The concentration ranges of the diluted samples cover the entire measuring range of the assays. Samples were assayed in 4-fold determination within a single run, on one instrument. SD and CVs were calculated for each 4-fold determination.

Reportable ranges were established as:

Elecsys sFlt-1 is 80-85000 pg/mL. .
. Elecsys PlGF is 10-5400 pg/mL.

Attention: For the Elecsys sFlt-1 assay, please use the PreciControl Multimarker controls with the lowest control level for sFlt-1 being approximately 100 pg/mL.

Analytical Sensitivity 4.3.

Limit of Blank (LoB) 4.3.1.

LoB of the Elecsys sFlt-1 and PlGF assays on the cobas e 411 analyzer was determined according to CLSI EP17-A2. Limit of Blank determines the highest observed measurement values for samples free of analyte. The Limit of Blank was determined as the 95th percentile of measurements of blank samples. The zero-level (blank) sample used was a human serum sample from pregnant female diluted with horse serum.

In total 60 determinations for analyte free samples have been obtained. Limit of Blank determines the highest observed measurement values for samples free of analyte. The Limit of Blank was determined as the 95th percentile of measurements of blank samples.

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Table 4. LoB for PIGF

Reagent lot	LoB (pg/mL)
1	0.732
2	0.946
3	0.482

All lots met the predetermined acceptance criterion. The LoB claim in the labeling will be set to ≤ 2 pg/mL.

Table 5. LoB for sFLT-1

Reagent lot	LoB (pg/mL)
1	4.00
2	3.51
3	2.92

All lots met the predetermined acceptance criterion. The LoB claim in the labeling will be set to 26.4 mg/dL in the samples can result in an up to 10 % decrease of the sFlt-1/PlGF ratio around the cutoff of 38 as shown in a spiking experiment.

Heterophilic human anti-mouse antibodies (HAMA) in human specimens can react with reagent antibodies, interfering with in vitro immunoassays. Patients routinely exposed to animals or animal serum products for diagnosis or therapies can be prone to this interference and anomalous values may be observed. Specimens from patients who have received mouse monoclonal antibodies for diagnosis or therapy may contain human anti-mouse antibodies and may interfere in assays that employ mouse monoclonal antibodies. Additional information may be required for diagnosis.

In rare cases, interference due to extremely high titers of antibodies to analyte-specific antibodies, streptavidin or ruthenium can occur. These effects are minimized by suitable test design. sFlt-1 and PlGF values determined on patient samples using other manufacturers' assays

20

may result in significantly different ratios, which could lead to wrong diagnostic conclusions. Therefore, only the Elecsys sFlt-1 and Elecsys PlGF assays should be used to calculate the ratio.

Analytical Specificity/Cross-Reactivity 4.7.

A cross-reactivity study was conducted with Elecsys sFit-1 and PIGF assays on the cobas e 411 analyzer to evaluate the potential cross-reacting compounds using human serum samples. Samples were measured in the presence and absence of the potential cross-reactants and crossreactivity was calculated with one lot of reagent.

Elecsys sFlt-1 assay

The monoclonal antibodies used are highly specific against human sFlt-1. The following cross-reactivities were found in samples with sFlt-1 levels between 1300 and 9000 pg/mL.

| Substance | Concentration tested
(pg/mL) | Cross-reactivity
(%) |
|-----------|---------------------------------|-------------------------|
| VEGFR2 | 11000000 | 500 U/L (see section Precautions and warnings).

13 additional pharmaceuticals and substances were tested with the Elecsys sFlt-1 and Elecsys PIGF assays. No interference with the Elecsys sFit-1 and Elecsys PIGF assays was found.

| Compound | Concentration
(mg/L) |
|-------------------|-------------------------|
| Albumin | 60000 |
| Caffeine | 59 |
| Calcium | 200 |
| Dihydralazin | 200 |
| Ethanol | 5 % (v/v) |
| Folic Acid | 2.4 |
| Gentamicin | 10 |
| Iron | 72 |
| Labetalol | 1.5 |
| Magnesium Sulfate | 252 |
| Methyldopa | 7.2 |
| Metroprolol | 5 |
| Nifedipine | 0.6 |

Table 14. Additional Substances Tested

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CLINICAL PERFORMANCE EVALUATION 5.

Praecis "Preeclampsia Risk Assessment: Evaluation of Cut-offs to Improve Stratification"

PRAECIS is a validation of the Roche sFlt-1/PlGF ratio cut-off of 38 as an aid in the risk assessment of pregnant women with a singleton pregnancy (23+0 to 34+6/7 weeks' gestation) with a hypertensive disorder of pregnancy to develop preeclampsia with severe features within two weeks from testing.

Study population included pregnant women (≥18 years of age) and gestational age 23+0 to 34+6/7 weeks who are admitted to the hospital with (or develop while hospitalized) a hypertensive disorder of pregnancy (preeclampsia, chronic hypertension with or without superimposed preeclampsia, or gestational hypertension).

Clinical Performance criteria of NPV and PPV were met for sFlt-1/PIGF ratio at a cutoff of 38 for the Intended Use and the Intended Use (Study Target) population. The primary study objective was fulfilled.

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| Assay | N of sPE a) | N of non-
sPE | Total N | Prevalence
(%) | Sensitivity
(%) (95% CI) | Specificity
(%) (95% CI) | NPV b)
(%) (95% CI) | Assay | N of sPE | N of non-sPE | Total N | PPV c)
(%) (95% CI) | Accuracy
(%) (95% CI) | LR- d)
(95% CI) | LR+ e)
(95% CI) |
|------------|-------------|------------------|---------|-------------------|-----------------------------|-----------------------------|-------------------------|------------|----------|--------------|---------|-------------------------|--------------------------|----------------------|----------------------|
| ratio > 38 | 170 | 84 | 254 | | 91.40 | 77.30 | 94.70 | ratio > 38 | 170 | 84 | 254 | | | | |
| ratio ≤ 38 | 16 | 286 | 302 | 33.45 | 91.40
(86.41, 95.00) | 77.30
(72.68, 81.47) | 94.70
(91.54, 96.94) | ratio ≤ 38 | 16 | 286 | 302 | 66.93
(60.77, 72.68) | 82.01
(78.56, 85.12) | 0.11
(0.07, 0.18) | 4.03
(3.32, 4.88) |
| Total | 186 | 370 | 556 | | | | | Total | 186 | 370 | 556 | | | | |

Table 15: Performance of the sFLT-1 and PIGF Assays in the Intended Use Population

sPE = preeclampsia with severe features a)

NPV = negative predictive value b)

PPV = positive predictive value c)

LR- = negative likelihood ratio d)

LR+ = positive likihood ratio e)

Table 16: Impact of hypertensive disorder at presentation on the prognostic performance of Roche Elecsys sFlt-1/PIGF ratio at cutoff of 38

| Hypertensive
disorders | n | Sensitivity
(%) (95% CI) | Specificity
(%) (95% CI) | PPV
(%) (95%
CI) | NPV
(%) (95%
CI) |
|------------------------------|-----|-----------------------------|-----------------------------|----------------------------|----------------------------|
| Preeclampsia | 142 | 95.06
(87.84, 98.64) | 72.13
(59.17, 82.85) | 81.91
(72.63,
89.10) | 91.67
(80.02,
97.68) |
| Superimposed
Preeclampsia | 68 | 93.75
(79.19, 99.23) | 55.56
(38.10, 72.06) | 65.22
(49.75,
78.65) | 90.91
(70.84,
98.88) |

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| Hypertensive
disorders | n | Sensitivity
(%) (95% CI) | Specificity
(%) (95% CI) | PPV
(%) (95%
CI) | NPV
(%) (95%
CI) |
|-----------------------------|-----|-----------------------------|-----------------------------|----------------------------|----------------------------|
| Chronic
Hypertension | 232 | 87.76
(75.23, 95.37) | 82.51
(76.22, 87.72) | 57.33
(45.38,
68.69) | 96.18
(91.87,
98.58) |
| Gestational
Hypertension | 114 | 83.33
(62.62, 95.26) | 78.89
(69.01, 86.79) | 51.28
(34.78,
67.58) | 94.67
(86.90,
98.53) |

Table 17: Impact of maternal age at presentation on the prognostic performance of Roche Elecsys sFlt-1/PIGF ratio at cutoff of 38

| Maternal
age | n | Sensitivity
(%) (95% CI) | Specificity
(%) (95% CI) | PPV
(%) (95% CI) | NPV
(%) (95% CI) |
|-----------------|-----|-----------------------------|-----------------------------|-------------------------|-------------------------|
|