← Product Code [PRI](/submissions/MI/subpart-d%E2%80%94serological-reagents/PRI) · K172713

# Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges, Lumipulse G B•R•A•H•M•S PCT Calibrators set (K172713)

_Fujirebio Diagnostics,Inc. · PRI · Dec 10, 2017 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PRI/K172713

## Device Facts

- **Applicant:** Fujirebio Diagnostics,Inc.
- **Product Code:** [PRI](/submissions/MI/subpart-d%E2%80%94serological-reagents/PRI.md)
- **Decision Date:** Dec 10, 2017
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3215
- **Device Class:** Class 2
- **Review Panel:** Microbiology

## Indications for Use

The Lumipulse G B•R•A•H•M•S PCT is a Chemiluminescent Enzyme Immunoassay (CLEIA) for the quantitative determination of PCT (procalcitonin) in human serum and plasma (sodium heparin, lithium heparin, sodium citrate or dipotassium EDTA) on the LUMIPULSE G System. Used in conjunction with other laboratory findings and clinical assessments, Lumipulse G B.R.A.H.M.S PCT is intended for use as an: · Aid in the risk assessment of critically ill patients on their first day of intensive care unit (ICU) admission for progression to severe sepsis and septic shock. · Aid in assessing the cumulative 28-day risk of all-cause mortality for patients diagnosed with severe sepsis or septic shock in the ICU or when obtained in the emergency department or other medical wards prior to ICU admission, using a change in PCT level over time. · Aid in decision making on antibiotic therapy for patients with suspected or confirmed lower respiratory tract infections (LRTI) - defined as community acquired pneumonia (CAP), acute bronchitis, and acute exacerbation of chronic obstructive pulmonary disease (AECOPD) – in an inpatient setting or an emergency department. · Aid in decision making on antibiotic discontinuation for patients with suspected or confirmed sepsis.

## Device Story

Lumipulse G B.R.A.H.M.S. PCT is an automated CLEIA assay for quantitative measurement of procalcitonin (PCT) in human serum/plasma. Used on the LUMIPULSE G1200 System; utilizes two-step sandwich immunoassay. Input: patient serum/plasma sample. Process: PCT binds to anti-PCT/anti-calcitonin monoclonal antibody-coated particles; unbound material washed; ALP-labeled anti-katacalcin antibody binds to immunocomplexes; substrate added; luminescence generated proportional to PCT concentration. Output: quantitative PCT concentration (ng/mL). Used in clinical/hospital laboratories by trained personnel. Results aid clinicians in sepsis risk assessment, mortality risk stratification, and antibiotic stewardship (initiation/discontinuation) for sepsis and lower respiratory tract infections. Benefits include standardized, automated PCT measurement to support critical care decision-making.

## Clinical Evidence

No clinical prospective/retrospective trials performed. Evidence consists of analytical performance studies (precision, linearity, LoD/LoQ, interference, cross-reactivity) and a method comparison study (n=223) against the predicate device. Method comparison showed high correlation (r=0.9846) and acceptable bias at clinical decision points. Reference interval study (n=213) established 95th percentile at 0.045 ng/mL.

## Technological Characteristics

CLEIA-based two-step sandwich immunoassay. Reagents: anti-PCT/anti-calcitonin monoclonal antibody-coated particles, alkaline phosphatase-labeled anti-katacalcin antibody. Energy source: LUMIPULSE G1200 System. Connectivity: Standalone instrument; web-based calculator for clinical decision support. Sterilization: Not applicable (reagents). Software: Automated analysis on LUMIPULSE G1200; web-based PCT change calculator.

## Regulatory Identification

A device to detect and measure non-microbial analyte(s) in human clinical specimens to aid in assessment of patients with suspected sepsis is identified as an in vitro device intended for the detection and qualitative and/or quantitative measurement of one or more non-microbial analytes in human clinical specimens to aid in the assessment of patients with suspected sepsis when used in conjunction with clinical signs and symptoms and other clinical and laboratory findings.

## Special Controls

A device to detect and measure non-microbial analyte(s) in human clinical specimens to aid in assessment of patients with suspected sepsis must comply with the following special controls:

*Classification.* Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include the device's detailed Indications for Use statement describing what the device detects and measures, the results provided to the user, whether the measure is qualitative and/or quantitative, the clinical indications for which the test is to be used, and the specific population(s) for which the device use is intended.
(2) Premarket notification submissions must include detailed documentation of the device description, including (as applicable), all device components, software, ancillary reagents required but not provided, explanation of the device principle and methodology, and for molecular devices include detailed documentation of the primer/probe sequence, design, and rationale for sequence selection.
(3) Premarket notification submissions must include detailed documentation of applicable analytical studies, such as, analytical sensitivity (Limit of Detection, Limit of Blank, and Limit of Quantitation), precision, reproducibility, analytical measuring range, interference, cross-reactivity, and specimen stability.
(4) Premarket notification submissions must include detailed documentation of a prospective clinical study or, if appropriate, results from an equivalent sample set. This detailed documentation must include the following information:
(i) Results must demonstrate adequate device performance relative to a well-accepted comparator.
(ii) Clinical sample results must demonstrate consistency of device output throughout the device measuring range likely to be encountered in the Intended Use population.
(iii) Clinical study documentation must include the original study protocol (including predefined statistical analysis plan), study report documenting support for the Indications for Use(s), and results of all statistical analyses.
(5) Premarket notification submissions must include evaluation of the level of the non-microbial analyte in asymptomatic patients with demographic characteristics (
*e.g.,* age, racial, ethnic, and gender distribution) similar to the Intended Use population.(6) As part of the risk management activities performed under 21 CFR 820.10(c) design and development, you must document an appropriate end user device training program that will be offered as part of your efforts to mitigate the risk of failure to correctly operate the instrument.
(7) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling, and a detailed explanation of the interpretation of the limitations of the samples (
*e.g.,* collected on day of diagnosis) must be included in the device's 21 CFR 809.10(b)(10) compliant labeling.

## Predicate Devices

- B•R•A•H•M•S PCT sensitive KRYPTOR® ([K171338](/device/K171338.md))

## Submission Summary (Full Text)

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>
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION MEMORANDUM

A. 510(k) Number:
K172713

B. Purpose for Submission:
To obtain a substantial equivalence determination for the Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges, Lumipulse G B•R•A•H•M•S PCT Calibrators set

C. Measurand:
Procalcitonin (PCT)

D. Type of Test:
Chemiluminescent Enzyme Immunoassay (CLEIA)

E. Applicant:
Fujirebio Diagnostics, Inc. (FDI)

F. Proprietary and Established Names:
Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges, Lumipulse G B•R•A•H•M•S PCT Calibrators set

G. Regulatory Information:

1. Regulation section:
21 CFR 866.3215; Device to detect and measure non-microbial analyte(s) in human clinical specimens to aid in assessment of patients with suspected sepsis

2. Classification:
Class II (Special Controls)

3. Product codes:
PRI, PMT, NTM

4. Panel:
83 - (Microbiology)

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H. Intended Use:

1. Intended use(s):

See Indications for Use.

2. Indication(s) for use:

Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges:

The Lumipulse G B•R•A•H•M•S PCT is a Chemiluminescent Enzyme Immunoassay (CLEIA) for the quantitative determination of PCT (procalcitonin) in human serum and plasma (sodium heparin, lithium heparin, sodium citrate or dipotassium EDTA) on the LUMIPULSE G System.

Used in conjunction with other laboratory findings and clinical assessments, Lumipulse G B•R•A•H•M•S PCT is intended for use as an:

- Aid in the risk assessment of critically ill patients on their first day of intensive care unit (ICU) admission for progression to severe sepsis and septic shock.
- Aid in assessing the cumulative 28-day risk of all-cause mortality for patients diagnosed with severe sepsis or septic shock in the ICU or when obtained in the emergency department or other medical wards prior to ICU admission, using a change in PCT level over time.
- Aid in decision making on antibiotic therapy for patients with suspected or confirmed lower respiratory tract infections (LRTI) – defined as community acquired pneumonia (CAP), acute bronchitis, and acute exacerbation of chronic obstructive pulmonary disease (AECOPD) – in an inpatient setting or an emergency department.
- Aid in decision making on antibiotic discontinuation for patients with suspected or confirmed sepsis.

Lumipulse G B•R•A•H•M•S PCT Calibrators set

Lumipulse G B•R•A•H•M•S PCT Calibrators set is for in vitro diagnostic use in the calibration of Lumipulse G B•R•A•H•M•S PCT on the LUMIPULSE G System.

3. Special conditions for use statement(s):

For prescription use.

4. Special instrument requirements:

All data were generated using the LUMIPULSE G1200 System.

I. Device Description:

Lumipulse G B•R•A•H•M•S PCT is an assay system, including a set of immunoassay reagents, for the quantitative measurement of PCT in specimens based on CLEIA technology by a two-step sandwich immunoassay method on the LUMIPULSE G1200 System.

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Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges: IRC 235058.

The Lumipulse G B•R•A•H•M•S PCT Immunoreaction Cartridges (IC) consists of 3 × 14 tests.

Each kit contains the following:

- Antibody-Coated Particle Solution
(Liquid when used, 250 µL/Immunoreaction Cartridge)
Contains 150 µg/mL anti-PCT monoclonal antibody (mouse) and anti-calcitonin monoclonal antibody (mouse) coated particles, protein stabilizers (bovine and mouse) and chemical stabilizers in 0.15 M sodium chloride/Tris buffer. This solution contains gelatin and turns into gel at 15°C or lower.
Preservative: Sodium azide

- Enzyme-Labeled Antibody Solution
(Liquid; 350 µL/Immunoreaction Cartridge)
Contains 0.25 µg/mL alkaline phosphatase (ALP: calf)-labeled anti-katacalcin monoclonal antibody (mouse), protein stabilizers (bovine, calf and mouse) and chemical stabilizers in 0.1 M sodium chloride/MES buffer.
Preservative: Sodium azide

Lumipulse G B•R•A•H•M•S PCT Calibrators set CAL SET 234150, Lyophilized, 2 × 2

Each calibrator kit contains one bottle each of Calibrators 1, 2, and Reconstituting Solution. These calibrators are lyophilized and are prepared by adding exactly 0.5 mL of Reconstituting Solution to each lyophilized calibrator. The calibrator kit is packaged separately.

- CAL 1: 0 ng/mL PCT calibrator (2 × 0.5 mL/vial)
- CAL 2: 100 ng/mL PCT calibrator (2 × 0.5 mL/vial); Contains procalcitonin in 0.15 M sodium chloride in Tris buffer with protein stabilizer (bovine).
Preservative: ProClin 300.
- RS Reconstituting Solution: Liquid, 1 × 10 mL
Preservative: Sodium azide

Materials Required - Not Provided:

- Control materials: Controls are not provided
NOTE: Lumipulse G B•R•A•H•M•S PCT Package insert indicates ‘Use control materials with at least two levels (e.g. low and high)’
Premarket studies used three Thermo Scientific MAS Omni•CARDIOControls (k122291).
o Level 1: 0.22 to 0.42 ng/mL
o Level 2: 1.49 to 2.77 ng/mL
o Level 3: 6.81 to 12.65 ng/mL

- Purified water
- Micropipettes
- Recommended sample cups; refer to the LUMIPULSE G System Operation Manual.

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J. Substantial Equivalence Information:

1. Predicate device name(s):
B•R•A•H•M•S PCT sensitive KRYPTOR

2. Predicate 510(k) number(s):
K171338

3. Comparison with predicate:

|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device
Lumipulse G
B•R•A•H•M•S PCT
K172713 | Predicate
B•R•A•H•M•S PCT sensitive
KRYPTOR
K171338  |
|  Device Type | In vitro diagnostic | Same  |
|  Classification | Class II | Same  |
|  Analyte | Procalcitonin | Same  |
|  Regulation | 21CFR § 866.3215;
Device to detect and measure
nonmicrobial analyte(s) in human
clinical specimens to aid in assessment
of patients with suspected sepsis | Same  |
|  Specimen
Collection
Method | Routine Phlebotomy
Techniques | Same  |
|  Indications for
Use | The Lumipulse G B•R•A•H•M•S PCT is
a Chemiluminescent Enzyme
Immunoassay (CLEIA) for the
quantitative determination of PCT
(procalcitonin) in human serum and
plasma (sodium heparin, lithium heparin,
sodium citrate or dipotassium EDTA) on
the LUMIPULSE G System.

Used in conjunction with other
laboratory findings and clinical
assessments, Lumipulse G
B•R•A•H•M•S PCT is intended for use
as an:
• Aid in the risk assessment of critically
ill patients on their first day of intensive
care unit (ICU) admission for
progression to severe sepsis and septic
shock.
• Aid in assessing the cumulative 28-day
risk of all-cause mortality for patients | The B·R·A·H·M·S PCT sensitive
KRYPTOR is an immunofluorescent
assay using Time-Resolved Amplified
Cryptate Emission (TRACE)
technology to determine the
concentration of PCT (procalcitonin) in
human serum and EDTA or heparin
plasma. The B·R·A·H·M·S PCT
sensitive KRYPTOR is intended to be
performed on the B·R·A·H·M·S
KRYPTOR analyzer family. Used in
conjunction with other laboratory
findings and clinical assessments.
B·R·A·H·M·S PCT sensitive
KRYPTOR is intended for use as
follows:
• to aid in the risk assessment of
critically ill patients on their first day of
ICU admission for progression to severe
sepsis and septic shock,
• to determine the change in PCT level  |

4

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|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device Lumipulse G B•R•A•H•M•S PCT K172713 | Predicate B•R•A•H•M•S PCT sensitive KRYPTOR K171338  |
|   | diagnosed with severe sepsis or septic shock in the ICU or when obtained in the emergency department or other medical wards prior to ICU admission, using a change in PCT level over time. • Aid in decision making on antibiotic therapy for patients with suspected or confirmed lower respiratory tract infections (LRTI) – defined as community acquired pneumonia (CAP), acute bronchitis, and acute exacerbation of chronic obstructive pulmonary disease (AECOPD) – in an inpatient setting or an emergency department. • Aid in decision making on antibiotic discontinuation for patients with suspected or confirmed sepsis. | over time as an aid in assessing the cumulative 28-day risk of all-cause mortality for patients diagnosed with severe sepsis or septic shock in the ICU or when obtained in the emergency department or other medical wards prior to ICU admission, • to aid in decision making on antibiotic therapy, for inpatients or patients in the emergency department with suspected or confirmed lower respiratory tract infections (LRTI) –defined as community-acquired pneumonia (CAP), acute bronchitis, and acute exacerbation of chronic obstructive pulmonary disease (AECOPD), • to aid in decision making on antibiotic discontinuation for patients with suspected or confirmed sepsis.  |
|  Differences  |   |   |
| --- | --- | --- |
|  Item | Device Lumipulse G B•R•A•H•M•S PCT K172713 | Predicate B•R•A•H•M•S PCT sensitive KRYPTOR K171338  |
|  Principle of Operation | Automated Quantitative Chemiluminescent Enzyme Immunoassay (CLEIA) | Time-Resolved Amplified Cryptate Emission (TRACE)  |
|  Instrument System | LUMIPULSE G System | BRAHMS KRYPTOR analyzer  |
|  Assay Type | Two-step sandwich immunoassay based on chemiluminescent technology | Immunofluorescent assay  |
|  Type of Specimen | Human serum and plasma (sodium heparin, lithium heparin, sodium citrate or dipotassium EDTA) | Human serum and plasma (EDTA, heparin)  |
|  Assay Range | 0.020 - 100 ng/mL | 0.02-5000ng/mL  |
|  Sample Volume | 60 μl | 50μl  |

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K. Standard/Guidance Document Referenced (if applicable):

- CLSI EP5-A3 - Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition
- CLSI EP7-A2 - Interference Testing in Clinical Chemistry; Approved Guideline-Second Edition
- CLSI EP28-A3c - Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline-Third Edition
- CLSI EP17-A2 - Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline Second Edition
- CLSI EP6-A - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline
- CLSI EP9-A3 – Measurement Procedure Comparison and Bias Estimation Using Patient Samples; approved Guideline – Third Edition
- CLSI EP25-A – Evaluation of Stability of In Vitro Diagnostic Reagents: Approved Guideline
- Guidance on Informed Consent for In Vitro Diagnostic Device Studies Using Leftover Human Specimens that are Not Individually Identifiable - Guidance for Sponsors, Institutional Review Boards, Clinical Investigators and FDA Staff Test Principle: Guidance for Industry and Food and Drug Administration Staff
- eCopy Program for Medical Device Submissions (December 31, 2012)
- Guidance for Industry and Food and Drug Administration Staff - Refuse to Accept Policy for 510(k)s (August 4, 2015)

L. Test Principle:

Lumipulse G B•R•A•H•M•S PCT is an assay system, including a set of immunoassay reagents, for the quantitative measurement of PCT in specimens based on CLEIA technology by a two-step sandwich immunoassay method on the LUMIPULSE G1200 System. PCT in specimens specifically binds to anti-PCT monoclonal antibody (mouse) and anti-calcitonin monoclonal antibody (mouse) on the particles, and antigen-antibody immunocomplexes are formed. The particles are washed and rinsed to remove unbound materials. Alkaline phosphatase (ALP; calf)-labeled anti-katacalcin monoclonal antibody (mouse) specifically binds to PCT of the immunocomplexes on the particles, and additional immunocomplexes are formed. The particles are washed and rinsed to remove unbound materials. Substrate Solution is added and mixed with the particles. AMPPD (3-(2’-Spiroadamantane)-4-Methoxy-4-(3’’-Phosphoryloxy) Phenyl-1, 2-Dioxetane Disodium Salt) contained in the Substrate Solution is dephosphorylated by the catalysis of ALP indirectly conjugated to particles. Luminescence (at a maximum wavelength of 477 nm) is generated by the cleavage reaction of dephosphorylated AMPPD. The luminescent signal reflects the amount of PCT.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Prior to the precision study, technicians were required to pass a 3-day (Lot A and B) and 1-day (Lot D) Familiarization Study to demonstrate proficiency on Lumipulse G PCT.

Eight panels (five native serum and three recombinant) were tested in duplicate. Thermo Scientific MAS Omni•CARDIO Controls were also tested for validity and precision. The results of the 20-day precision calculations for Lumipulse G B•R•A•H•M•S PCT performed at

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Fujirebio Diagnostics, Inc are shown below:

Lot A Immunoreaction Cartridges (ICs)/Lot A Calibrators for Control Levels 1-3 and Panels 1-6
Lot D ICs/Lot D Calibrators for Panels 7 and 8

The analyses determined the CV for the within laboratory precision of the Lumipulse G B•R•A•H•M•S PCT for the eight (8) panels CV ranged from 1.8% to 3.1%. The CV for the within laboratory precision of the Lumipulse G B•R•A•H•M•S PCT for the three (3) controls ranged from 3.3% to 4.7%. The precision of all controls and panels for the 20-day Precision Study met the acceptance criteria of a CV ≤ 10%. The results of the 20-day precision calculations for Lumipulse G B•R•A•H•M•S PCT are shown in Table 1 below:

Table 1: Summary for FDI 20-day Precision (n=80 for each sample) for Lot A (Controls and Panels 1-6) and Lot D (Panels 7 and 8)

|   |   | Within-Run (Repeatability) |   | Between Run |   | Between-Day |   | Within-Laboratory (Total)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample | Mean (ng/mL) | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Control Level 1 | 0.662 | 0.010 | 1.6 | 0.026 | 3.9 | 0.014 | 2.1 | 0.031 | 4.7  |
|  Control Level 2 | 3.911 | 0.045 | 1.2 | 0.129 | 3.3 | 0.026 | 0.7 | 0.139 | 3.6  |
|  Control Level 3 | 14.978 | 0.163 | 1.1 | 0.393 | 2.6 | 0.245 | 1.6 | 0.490 | 3.3  |
|  Panel 1 | 0.300 | 0.005 | 1.6 | 0.004 | 1.5 | 0.003 | 1.0 | 0.007 | 2.4  |
|  Panel 2 | 2.219 | 0.032 | 1.5 | 0.016 | 0.7 | 0.027 | 1.2 | 0.045 | 2.0  |
|  Panel 3 | 12.323 | 0.129 | 1.0 | 0.144 | 1.2 | 0.098 | 0.8 | 0.217 | 1.8  |
|  Panel 4 | 36.217 | 0.379 | 1.0 | 0.308 | 0.8 | 0.445 | 1.2 | 0.661 | 1.8  |
|  Panel 5 | 54.132 | 0.596 | 1.1 | 0.529 | 1.0 | 0.642 | 1.2 | 1.024 | 1.9  |
|  Panel 6 | 80.361 | 1.265 | 1.6 | 0.879 | 1.1 | 0.632 | 0.8 | 1.666 | 2.1  |
|  Panel 7 | 0.112 | 0.002 | 2.0 | 0.002 | 2.2 | 0.001 | 1.1 | 0.004 | 3.1  |
|  Panel 8 | 0.534 | 0.010 | 1.9 | 0.008 | 1.5 | 0.003 | 0.6 | 0.013 | 2.5  |

# Lot-To-Lot Reproducibility for Combined Data

The precision analyses for the combined lot-to-lot analysis for Lots A, B and C determined the CV for the within laboratory reproducibility for Lumipulse G B•R•A•H•M•S PCT to be ≤ 5.3% in this study. The CV for the within laboratory reproducibility for Lumipulse G B•R•A•H•M•S PCT was determined to be ≤ 3.0% for the panels 1-6. The CV for the within laboratory reproducibility for Lumipulse G B•R•A•H•M•S PCT was determined to be ≤ 5.3% for the three (3) controls. The CV for the within laboratory reproducibility of Lumipulse G B•R•A•H•M•S PCT ranged from 1.8% to 5.3%. The CV for the between-lot reproducibility for Lumipulse G B•R•A•H•M•S PCT was ≤ 6.5%. See Table 2 below.

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Table 2: Summary of the Lot-to-Lot Reproducibility for the Combined Data for Lots A, B, C (n=120 for each sample)

|   |   | Between Lots |   | Between Day |   | Between Run |   | Within Runs (Repeatability) |   | Within-Laboratory (Total)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample | Mean (ng/L) | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Control Level 1 | 0.634 | 0.030 | 4.7 | 0.013 | 2.0 | 0.030 | 4.7 | 0.009 | 1.4 | 0.033 | 5.3  |
|  Control Level 2 | 3.816 | 0.137 | 3.6 | 0.016 | 0.4 | 0.127 | 3.3 | 0.055 | 1.4 | 0.139 | 3.7  |
|  Control Level 3 | 14.875 | 0.313 | 2.1 | 0.141 | 0.9 | 0.434 | 2.9 | 0.164 | 1.1 | 0.487 | 3.3  |
|  Panel 1 | 0.290 | 0.012 | 4.3 | 0.004 | 1.3 | 0.006 | 2.0 | 0.005 | 1.8 | 0.009 | 3.0  |
|  Panel 2 | 2.163 | 0.075 | 3.4 | 0.021 | 1.0 | 0.038 | 1.8 | 0.026 | 1.2 | 0.048 | 2.2  |
|  Panel 3 | 12.332 | 0.320 | 2.6 | 0.221 | 1.8 | 0.133 | 1.1 | 0.122 | 1.0 | 0.280 | 2.3  |
|  Panel 4 | 38.147 | 2.280 | 6.0 | 0.363 | 1.0 | 0.467 | 1.2 | 0.367 | 1.0 | 0.701 | 1.8  |
|  Panel 5 | 57.114 | 3.392 | 5.9 | 0.856 | 1.5 | 0.625 | 1.1 | 0.674 | 1.2 | 1.248 | 2.2  |
|  Panel 6 | 86.233 | 5.585 | 6.5 | 1.018 | 1.2 | 1.138 | 1.3 | 1.489 | 1.7 | 2.141 | 2.5  |

# Site to Site Reproducibility for Combined Data

The reproducibility analyses for the combined site-to-site analysis for Lot A determined the CV for the total reproducibility for Lumipulse G B•R•A•H•M•S PCT to be  $\leq 4.9\%$  in this study. The CV for the reproducibility for Lumipulse G B•R•A•H•M•S PCT was determined to be  $\leq 4.8\%$  for the six (6) panels. The CV for the reproducibility for Lumipulse G B•R•A•H•M•S PCT was determined to be  $\leq 4.9\%$  for the three (3) controls. The CV for the reproducibility of Lumipulse G B•R•A•H•M•S PCT ranged from  $2.9\%$  to  $4.9\%$ . The CV for the between site reproducibility for Lumipulse G B•R•A•H•M•S PCT was  $\leq 4.7\%$ . Results are summarized in Table 3 below.

Table 3: Summary of the Site-to-Site Reproducibility for the Combined Data for Lot A (n=120 for each panel)

|   |   | Between Sites |   | Between Days |   | Between Runs |   | Within Runs (Repeatability) |   | Reproducibility (Total)  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample | Mean (ng/mL) | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Control Level 1 | 0.647 | 0.019 | 2.9% | 0.023 | 3.5% | 0.020 | 3.0% | 0.012 | 1.8% | 0.032 | 4.9%  |
|  Control Level 2 | 3.872 | 0.077 | 2.0% | 0.109 | 2.8% | 0.096 | 2.5% | 0.054 | 1.4% | 0.149 | 3.8%  |
|  Control Level 3 | 15.062 | 0.085 | 0.6% | 0.337 | 2.2% | 0.312 | 2.1% | 0.166 | 1.1% | 0.481 | 3.2%  |
|  Panel 1 | 0.291 | 0.008 | 2.8% | 0.008 | 2.7% | 0.010 | 3.6% | 0.005 | 1.9% | 0.014 | 4.8%  |
|  Panel 2 | 2.170 | 0.033 | 1.5% | 0.049 | 2.3% | 0.061 | 2.8% | 0.032 | 1.5% | 0.075 | 3.5%  |
|  Panel 3 | 12.297 | 0.375 | 3.0% | 0.222 | 1.8% | 0.417 | 3.4% | 0.170 | 1.4% | 0.485 | 3.9%  |
|  Panel 4 | 37.074 | 1.303 | 3.5% | 0.586 | 1.6% | 1.182 | 3.2% | 0.361 | 1.0% | 1.354 | 3.6%  |
|  Panel 5 | 55.428 | 2.043 | 3.7% | 1.144 | 2.1% | 1.385 | 2.5% | 0.613 | 1.1% | 1.846 | 3.3%  |
|  Panel 6 | 82.850 | 3.868 | 4.7% | 1.006 | 1.2% | 1.719 | 2.1% | 1.285 | 1.6% | 2.365 | 2.9%  |

# b. Linearity/assay reportable range:

Dilutions targeted the top of the measurement range of the Lumipulse G B•R•A•H•M•S PCT assay ( $\sim 120 \, \mathrm{ng/mL}$ ) and went below the LOQ ( $< 0.020 \, \mathrm{ng/mL}$ ). Eleven separate dilutions of the High Serum Pool (HSP) by the Low Serum Pool (LSP) were prepared. Undiluted High and Low serum pools were tested as sample 1 and sample 13 in the dilution series Lumipulse G

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B•R•A•H•M•S PCT on the LUMIPULSE G1200 System demonstrated linearity in a study consistent with the guidelines in the CLSI Protocol EP6-A.9) High and low sample pools were created using patient serum samples that contained naturally expressed PCT. Lumipulse G B•R•A•H•M•S PCT assay was shown to be linear within the actual test results of 0.010 ng/mL to 104.260 ng/mL and supports the claimed Lumipulse G B•R•A•H•M•S PCT measurement range of 0.020 ng/mL – 100.000 ng/mL. Results are summarized in Table 4 below.

Table 4: Linearity
|  Dilution | Target value (ng/mL) | Mean measured value (ng/mL) | SD (ng/mL) | CV %  |
| --- | --- | --- | --- | --- |
|  1 | 120 | 104.260* | 2.990 | 2.9  |
|  2 | 100 | 86.680 | 0.529 | 0.6  |
|  3 | 80 | 70.449 | 0.693 | 1.0  |
|  4 | 60 | 53.370 | 0.347 | 0.7  |
|  5 | 40 | 35.961 | 0.100 | 0.3  |
|  6 | 20 | 17.555 | 0.617 | 3.5  |
|  7 | 10 | 8.015 | 0.125 | 1.6  |
|  8 | 5 | 4.087 | 0.085 | 2.1  |
|  9 | 2 | 1.613 | 0.011 | 0.7  |
|  10 | 0.481 | 0.424 | 0.009 | 2.0  |
|  11 | 0.241 | 0.228 | 0.002 | 0.8  |
|  12 | 0.121 | 0.111 | 0.001 | 1.3  |
|  13 | 0.001 | 0.010 | 0.001 | 6.1  |

The statistical evaluation of linearity was determined using EP6-A. Linear, quadratic and cubic regressions were fitted. Lumipulse G B•R•A•H•M•S PCT correlated with expected concentrations according to the linear regression formula: $y = 0.008734 + 0.856577x$; $R^2 = 0.9979$.

## Dilution Verification

A dilution verification study was performed to compare and assess manual dilution and automated (on-board) dilution for Lumipulse G B•R•A•H•M•S PCT. Three specimens (neat, manually diluted 1:10, manually diluted 1:32, automated 1:10) were tested in triplicate. Manual dilutions performed at 1:32 are not recommended for Lumipulse G B·R·A·H·M·S PCT. Automated dilutions for 1:100, 1:200, and 1:1000 on the LUMIPULSE G1200 System can cause erroneous results and should not be used when a specimen exceeds the 100 ng/mL.

Manual dilutions and automated dilutions performed at 1:10 are recommended for Lumipulse G B•R•A•H•M•S PCT.

## Spike and Recovery

Three individual apparently healthy serum samples were spiked and compared to the diluent with the identical concentration. Percent recovery was calculated as follows for individual spiked samples and was found to be within $100 \pm 10\%$ for all samples tested. See study results in Table 5 below.

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- % Recovery = (Mean Measured Natural Test Sample Matrix Concentration (ng/mL) - Endogenous Sample Concentration/Mean Measured Specimen Diluent 1 Concentration (ng/mL)) x 100%

Table 5: Spike and Recovery.

|   | Sample ID | Sample Mean (ng/ml) | Diluent (ng/ml) | Recovery (%)  |
| --- | --- | --- | --- | --- |
|  Serum Sample 1 | Endogenous | 0.011 | 0 |   |
|   |  2 | 0.097 | 0.091 | 95  |
|   |  3 | 0.223 | 0.223 | 95  |
|   |  4 | 0.445 | 0.455 | 95  |
|   |  5 | 1.67 | 1.719 | 97  |
|   |  6 | 8.885 | 9.073 | 98  |
|   |  7 | 30.317 | 31.553 | 96  |
|   |  8 | 44.417 | 45.649 | 97  |
|   |  9 | 63.408 | 63.252 | 100  |
|   |  10 | 86.949 | 82.587 | 105  |
|  Serum Sample 2 | Endogenous | 0.016 | 0.000 |   |
|   |  2 | 0.099 | 0.091 | 91  |
|   |  3 | 0.22 | 0.223 | 91  |
|   |  4 | 0.433 | 0.455 | 92  |
|   |  5 | 1.684 | 1.719 | 97  |
|   |  6 | 8.83 | 9.073 | 97  |
|   |  7 | 28.963 | 31.553 | 92  |
|   |  8 | 45.224 | 45.649 | 99  |
|   |  9 | 62.08 | 63.252 | 98  |
|   |  10 | 86.22 | 82.587 | 104  |
|  Serum Sample 3 | Endogenous | 0.008 | 0.000 |   |
|   |  2 | 0.096 | 0.091 | 97  |
|   |  3 | 0.225 | 0.223 | 97  |
|   |  4 | 0.44 | 0.455 | 95  |
|   |  5 | 1.661 | 1.719 | 96  |
|   |  6 | 8.75 | 9.073 | 96  |
|   |  7 | 30.363 | 31.553 | 96  |
|   |  8 | 45.014 | 45.649 | 99  |
|   |  9 | 61.666 | 63.252 | 97  |
|   |  10 | 84.661 | 82.587 | 103  |

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Traceability:

Lumipulse G B·R·A·H·M·S PCT values are expressed as ng/mL, and are related to a Fujirebio Inc. maintained reference preparation. The calibrators for the Lumipulse G B·R·A·H·M·S PCT product are manufactured gravimetrically and are traceable to B·R·A·H·M·S PCT sensitive Kryptor.

Master calibration data is recorded in a two-dimensional bar code on the Lumipulse G B·R·A·H·M·S PCT Immunoreaction Cartridge case. The calibration curve is created based on recorded master calibration data and the calibration data. The PCT concentration of a specimen

{10}

is automatically calculated from the calibration curve. The result of the calculation is reported in ng/mL.

Recalibration is required after a pre-determined calibration interval, when 30 days have elapsed since the previous calibration; quality control results fall out of range or when a new Lumipulse G B·R·A·H·M·S PCT Immunoreaction Cartridge lot is loaded.

## Reagent Stability:

The shelf life for Lumipulse G B·R·A·H·M·S PCT Immunoreaction Cartridges and the Lumipulse G B·R·A·H·M·S PCT Calibrators is 6 months at 2–10°C. The shelf-life stability protocols and acceptance criteria were reviewed and found to be acceptable.

## On-Board Immunoreaction Cartridge Stability Study

The shelf life for Lumipulse G B·R·A·H·M·S PCT Immunoreaction Cartridges is 6 months at 2–10°C. The Lumipulse G B·R·A·H·M·S PCT Cartridges can be stored on-board the LUMIPULSE G System for a maximum of 30 days.

## On Board Sample Stability Study

A study was performed to establish stability claims for the stability of samples on-board the LUMIPULSE G1200 instrument using Lumipulse G B·R·A·H·M·S PCT. Samples have an on-board stability up to 3 hours.

## Reagent Transport Conditions

Lumipulse G B·R·A·H·M·S PCT Immunoreaction Cartridges and the Lumipulse G B·R·A·H·M·S PCT Calibrators are shipped at 2–10°C. Materials are shipped to the end user using an insulated container and a predetermined configuration of gel (cold and/or frozen) packs to maintain the product for up to 72 hours when stored at ambient temperature.

## Sample Stability Study

The purpose of this study was to establish the stability of serum and plasma samples for Lumipulse G B·R·A·H·M·S PCT. The following information regarding sample stability are provided in Table 6 and will be included in the final labeling.

Table 6: Sample Stability
|  Tube | Condition 1 (25°C ±2°C) | Condition 2 (2°C-10°C) | Condition 3 (25°C ±2°C *)  |
| --- | --- | --- | --- |
|  SST | 7 Hours | 2 Days | 3 Hours  |
|  Red Top | 4 Hours | 1 Day | 7 Hours  |
|  K_{2}EDTA, Lithium Heparin, Sodium Heparin, and Sodium Citrate | 1 Day | 2 Days | 8 Hours  |

*Samples were tested after the draw (after introduction of PCT spike) while kept on-the-clot/separator/cells.

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12

# Sample Freeze Thaw Stability Study

A study to establish the maximum number of freeze thaw cycles acceptable for samples being measured by Lumipulse G B·R·A·H·M·S PCT assay was performed. Specimens can be frozen and thawed up to and including 3 freeze thaw cycles for the Lumipulse G B·R·A·H·M·S PCT for the following tube types: SST Serum, Red Top Serum, K₂EDTA Plasma, Sodium Heparin Plasma, Lithium Heparin Plasma, and Sodium Citrate Plasma.

## d. Detection limit:

The LoB, LoD and LoQ were determined using methods consistent with the guidelines in the CLSI protocol EP17-A2.

The LoB for Lumipulse G B·R·A·H·M·S PCT was 0.0095 ng/mL. The LoD for Lumipulse G B·R·A·H·M·S PCT on the LUMIPULSE G1200 System was 0.0114 ng/mL. Seven low level specimens were tested over three days using two LUMIULSE G1200 Systems and two Lumipulse G B·R·A·H·M·S PCT lots for a total of 120 determinations per panel.

The LoQ for Lumipulse G B·R·A·H·M·S PCT on the LUMIPULSE G1200 System was 0.0114 ng/mL. The % Total Error (TE), % bias and %CV were calculated:

- TE ≤ 11.4% at 0.25 ng/mL with a % bias ≤ -5.7% and a precision CV ≤ 2.8%.
- TE ≤ 14.1% at 0.10 ng/mL with a % bias ≤ -9.2% and a precision CV ≤ 2.5%.

## e. High Dose Hook Effect

For Lumipulse G B·R·A·H·M·S PCT on the LUMIPULSE G1200 System, no high dose effect was observed for samples containing approximately 12,000 ng/mL of PCT.

## f. Analytical specificity:

### Interference Testing:

Lumipulse G B·R·A·H·M·S PCT on the LUMIPULSE G1200 System demonstrated an average interference of ≤10% (for each compound) in a study consistent with the guidelines in the CLSI Protocol EP7-A2. Human serum specimen pools with procalcitonin concentrations of approximately 0.25 and 2.0 ng/mL were supplemented with potentially interfering compounds. The following compounds were tested using Lumipulse G B·R·A·H·M·S PCT and found not to interfere with the assay. See Tables 7 and 8 below.

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Table 7: Endogenous Interferents

|  Endogenous Interferent | Test Concentration  |
| --- | --- |
|  Free Bilirubin(unconjugated) | 50 mg/dL  |
|  Conjugated Bilirubin | 50 mg/dL  |
|  Hemoglobin | 400 mg/dL  |
|  Total Protein (Human Serum Albumin) | 12 g/dL  |
|  Triglycerides (Intralipid 20% Emulsion) | 2500 mg/dL  |
|  Immunoglobulin G (IgG) | 5 g/dL  |
|  Biotin | 19.7 mg/dL  |
|  Human Anti-Mouse Antibodies (HAMA) | 1,000 ng/mL  |
|  Rheumatoid Factor (RF) | 1,000 IU/mL  |

Table 8: Exogenous Interferent

|  Exogenous Interferent | Concentration (mg/dL)  |
| --- | --- |
|  Acetaminophen | 20.01  |
|  Acetylsalicylic Acid | 65.22  |
|  1% Ethanol | 789  |
|  Azithromycin | 2  |
|  Caffeine | 6  |
|  Celecoxib | 24  |
|  Cetirizine HCl | 0.55  |
|  Dextromethorphan | 0.1  |
|  Dobutamine | 1.13  |
|  Dopamine | 13  |
|  Doxycycline | 5  |
|  Epinephrine | 0.18  |
|  Furosemide | 2  |
|  Heparin | 8000  |
|  Ibuprofen | 50.03  |
|  Imipenem | 118  |
|  Levofloxacin | 2.93  |
|  Loratadine | 0.05  |
|  Nicotine | 0.1  |
|  Noradrenaline (norepinephrine) | 0.2  |
|  Oxymetazoline HCl | 0.09  |
|  Phenylephrine | 6  |
|  Prednisolone | 0.3  |
|  Salmeterol | 0.006  |
|  Tiotropium | 0.0022  |
|  Vancomycin | 300  |

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14

# Cross Reactivity:

Lumipulse G B•R•A•H•M•S PCT on the LUMIPULSE G1200 System was evaluated for cross-reactivity of the assay with other substances that are similar in structure to PCT in a study consistent with the guidelines in the CLSI Protocol EP7-A2. Human serum specimens with PCT concentrations of approximately 0.1, 0.25, 0.5, 2.0 and 80 ng/mL (serum pool 1, 2, 3, 4 and 5, respectively) were supplemented with potentially cross-reacting compounds. The compounds were tested at the concentrations listed below and found to have the following percent cross-reactivity. The % cross-reactivity for each individual sample was calculated using the following equation. See results in Table 9 below.

- % Cross Reactivity = (Mean Concentration$_{\text{Test}}$ (ng/mL) – Mean Concentration$_{\text{Control}}$ (ng/mL) / Concentration$_{\text{Cross Reactant}}$ (ng/mL)) X 100

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Table 9: Cross Reactivity

|  Interferent | Sample ID | Mean Test Concentration (ng/mL) (n=3) | Mean Control Concentration (ng/mL) (n=3) | % Cross Reactivity  |
| --- | --- | --- | --- | --- |
|  Human Calcitonin 10 ng/mL | Serum Pool 1 | 0.113 | 0.115 | -0.020%  |
|   |  Serum Pool 2 | 0.263 | 0.265 | -0.020%  |
|   |  Serum Pool 3 | 0.537 | 0.532 | 0.050%  |
|   |  Serum Pool 4 | 2.111 | 2.014 | 0.970%  |
|   |  Serum Pool 5 | 77.004 | 77.275 | -2.710%  |
|  Human Katacalcin 10 ng/mL | Serum Pool 1 | 0.113 | 0.116 | -0.030%  |
|   |  Serum Pool 2 | 0.268 | 0.266 | 0.020%  |
|   |  Serum Pool 3 | 0.537 | 0.541 | -0.040%  |
|   |  Serum Pool 4 | 2.118 | 2.134 | -0.160%  |
|   |  Serum Pool 5 | 76.476 | 76.417 | 0.590%  |
|  α-CGRP 10,000 ng/mL | Serum Pool 1 | 0.113 | 0.113 | 0.000%  |
|   |  Serum Pool 2 | 0.261 | 0.263 | 0.000%  |
|   |  Serum Pool 3 | 0.525 | 0.530 | 0.000%  |
|   |  Serum Pool 4 | 2.126 | 2.131 | 0.000%  |
|   |  Serum Pool 5 | 78.339 | 77.526 | 0.008%  |
|  β-CGRP 10,000 ng/mL | Serum Pool 1 | 0.117 | 0.115 | 0.000%  |
|   |  Serum Pool 2 | 0.269 | 0.265 | 0.000%  |
|   |  Serum Pool 3 | 0.540 | 0.534 | 0.000%  |
|   |  Serum Pool 4 | 2.173 | 2.175 | 0.000%  |
|   |  Serum Pool 5 | 77.705 | 77.443 | 0.003%  |
|  Salmon Calcitonin 13.2 μg/mL | Serum Pool 1 | 0.116 | 0.111 | 0.000%  |
|   |  Serum Pool 2 | 0.265 | 0.272 | 0.000%  |
|   |  Serum Pool 3 | 0.537 | 0.537 | 0.000%  |
|   |  Serum Pool 4 | 2.185 | 2.157 | 0.000%  |
|   |  Serum Pool 5 | 77.137 | 77.801 | -0.005%  |
|  Eel Calcitonin 7.5 μg/mL | Serum Pool 1 | 0.115 | 0.114 | 0.000%  |
|   |  Serum Pool 2 | 0.264 | 0.263 | 0.000%  |
|   |  Serum Pool 3 | 0.542 | 0.525 | 0.000%  |
|   |  Serum Pool 4 | 2.172 | 2.141 | 0.000%  |
|   |  Serum Pool 5 | 77.631 | 78.072 | -0.006%  |

f. Assay cut-off: See clinical cut-off

2. Comparison studies:

a. Method comparison with predicate device:

The Lumipulse G B•R•A•H•M•S PCT method comparison was performed on the LUMIPULSE G1200 System in a study consistent with the guidelines in CLSI Protocol EP09-A3C.

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B•R•A•H•M•S PCT sensitive KRYPTOR testing was performed following the appropriate QC procedures and manufacturer's instruction specified in the B•R•A•H•M•S PCT sensitive KRYPTOR package insert.

Sample testing with Lumipulse G B•R•A•H•M•S PCT was performed on three LUMIPULSE G1200 Systems utilizing two lots of ICs, one lot of calibrators, and four calibrator curves. There were 102 samples from female subjects (45.7%) and 121 samples from male subjects (54.3%) in the method comparison sample set. The PCT range of samples tested and age demographics are found in Tables 10 and 11 below.

Table 10: Total Samples per Specified Lumipulse G B•R•A•H•M•S PCT Ranges

|  Lumipulse G B•R•A•H•M•S PCT Ranges | Number of Samples  |
| --- | --- |
|  0.000-0.250 ng/mL | 53  |
|  0.251-0.500 ng/mL | 35  |
|  0.501-2.000 ng/mL | 47  |
|  2.001-10.000 ng/mL | 32  |
|  10.001-20.000 ng/mL | 11  |
|  20.001-50.000 ng/mL | 27  |
|  50.001-100.000 ng/mL | 10  |
|  100.001-250.000 ng/mL | 7  |
|  250.001-1000.000 ng/mL | 1  |
|  TOTAL | 223  |

Table 11: Summary of Age Demographics for Evaluable Subjects

|  Age Group (years) | Evaluable Subjects  |
| --- | --- |
|  22-29 | 2  |
|  30-39 | 9  |
|  40-49 | 14  |
|  50-59 | 21  |
|  60-69 | 63  |
|  70-79 | 52  |
|  80-89 | 44  |
|  90-99 | 18  |
|  Total | 223  |
|  Mean Age | 69.3  |
|  Median Age | 70.0  |
|  Standard Deviation (SD) (years) | 14.9  |
|  Minimum Age | 26  |
|  Maximum Age | 98  |

Two separate analysis of the study data were performed, one with analysis restricted to samples with B•R•A•H•M•S PCT sensitive KRYPTOR assays no more than 50.0 ng/mL and one with an extended analysis including all samples tested. Weighted Deming and Passing-Bablok regressions were performed with slope, correlation coefficient, intercept and mean difference along with 95% CIs determined and found to be acceptable. Bland-Altman Plots with mean % difference, percent limits of agreement and 95% CIs for Lumipulse G PCT and the predicate with B•R•A•H•M•S PCT sensitive KRYPTOR were also calculated and found to be acceptable.

16

{16}

Analysis of Samples within the Predicate Measurement Range:

207 were samples tested with Lumipulse G B•R•A•H•M•S PCT had results ranging from 0.054-58.156 ng/mL with B•R•A•H•M•S PCT sensitive KRYPTOR results for these same samples ranged from 0.024-48.430 ng/mL. Weighted Deming Regression results are found in Table 12. The clinical bias and confidence intervals (CIs) at the clinical decision points are listed in Table 13.

Table 12: Weighted Deming Regression for Lumipulse G B•R•A•H•M•S PCT when compared to B•R•A•H•M•S PCT sensitive KRYPTOR (measurement range 0.020-50.000 ng/mL)

|  n | Correlation Coefficient (r) | Intercept (95% CI) | Slope (95% CI) | Mean Difference (ng/mL) |   |
| --- | --- | --- | --- | --- | --- |
|  207 | 0.9535 | -0.0044 (-0.0223, 0.0135) | 1.0199 (0.9633, 1.0765) | 0.185 |   |

Table 13: Clinical bias and CIs at the clinical decision points

|  Clinical Decision Point (ng/mL) | Bias (ng/mL) | Lower CI | Upper CI  |
| --- | --- | --- | --- |
|  0.100 | -0.0031 | -0.0180 | 0.0117  |
|  0.250 | -0.0009 | -0.0148 | 0.0130  |
|  0.500 | 0.0027 | -0.0185 | 0.0239  |
|  2.000 | 0.0246 | -0.0749 | 0.1242  |

From the total 207 individual lithium heparin plasma samples tested, the percent agreement between the Lumipulse G B•R•A•H•M•S PCT and B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off  $0.500\mathrm{ng / mL}$  was determined. See Table 14 for  $2\times 2$  summary table.

- Negative percent agreement (NPA):  $95.5\%$  (84/88),  $95\%$  CI:  $88.9\%$ ,  $98.2\%$
- Positive percent agreement (PPA):  $96.6\%$  (115/119),  $95\%$  CI:  $91.7\%$ ,  $98.7\%$

Table 14: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR at 0.500 ng/mL cut-off

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |
| --- | --- | --- | --- |
|   |  ≤0.500 | >0.500 | Total  |
|  ≤0.500 | 84 | 4 | 88  |
|  >0.500 | 4 | 115 | 119  |
|  Total | 88 | 119 | 207  |

{17}

From the total 207 individual lithium heparin plasma samples tested, the percent agreement between the Lumipulse G B•R•A•H•M•S PCT and B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off  $2.000\mathrm{ng / mL}$  was determined. See Table 15, 16 and 18 for  $2\times 2$ ,  $3\times 3$  summary and regression analyses, respectively. Additionally, Table 17 presents a cross-tabulation of all samples within each medical decision point range.

- NPA:  $99.2\%$  (132/133),  $95\%$  CI:  $95.9\%$ ,  $99.9\%$
- PPA:  $95.9\%$  (71/74),  $95\%$  CI:  $88.7\%$ ,  $98.6\%$

Table 15: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR at  $2.000\mathrm{ng / mL}$  cut-off

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |
| --- | --- | --- | --- |
|   |  ≤2.000 | >2.000 | Total  |
|  ≤2.000 | 132 | 1 | 133  |
|  >2.000 | 3 | 71 | 74  |
|  Total | 135 | 72 | 207  |

Table 16: 3x3 Table: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |   |
| --- | --- | --- | --- | --- |
|   |  ≤0.500 | 0.501<PCT≤2.000 | >2.000 | Total  |
|  0.500 | 84 | 4 | 0 | 88  |
|  0.501<PCT≤2.000 | 4 | 40 | 1 | 45  |
|  >2.000 | 0 | 3 | 71 | 74  |
|  Total | 88 | 47 | 72 | 207  |

Table 17: Cross Tabulation of all Samples (n=207) within each Medical Decision Point Range

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/m)L | Lumipulse G PCT (ng/mL)  |   |   |   |   |
| --- | --- | --- | --- | --- | --- |
|   |  0.0237-0.100 | 0.101-0.250 | 0.251-0.500 | 0.501-2.000 | 2.001-58.200  |
|  0.0237-0.100 | 4 | 5 | 0 | 0 | 0  |
|  0.101-0.250 | 6 | 32 | 5 | 2 | 0  |
|  0.251-0.500 | 0 | 6 | 26 | 2 | 0  |
|  0.501-2.000 | 0 | 0 | 4 | 40 | 1  |
|  2.001-58.200 | 0 | 0 | 0 | 3 | 71  |

Table 18: Regression Analyses (N=207)

|   | Parameter | Estimate | Lower CI | Upper CI  |
| --- | --- | --- | --- | --- |
|  Weighted Deming | Intercept | -0.0044 | -0.0223 | 0.0135  |
|   |  Slope | 1.0199 | 0.9633 | 1.0765  |
|  Passing- Bablok | Intercept | -0.007 | -0.025 | 0.004  |
|   |  Slope | 1.000 | 0.981 | 1.038  |

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Analysis of Samples with the Lumipulse G B•R•A•H•M•S PCT extended measurement range:

223 samples tested with Lumipulse G B•R•A•H•M•S PCT had results ranging from 0.054-348.940 ng/mL and B•R•A•H•M•S PCT sensitive KRYPTOR results for these same samples ranged from 0.024-345.900 ng/mL. Weighted Deming Regression results are found in Table 19. The clinical bias and CIs at the clinical decision points were determined. No evidence of bias was found at the medical decision points. (See results in Table 20.)

Table 19: Weighted Deming Regression for Lumipulse G B•R•A•H•M•S PCT when compared to B•R•A•H•M•S PCT sensitive KRYPTOR (measurement range &gt;0.020 ng/mL)

|  n | Correlation Coefficient (r) | Intercept (95% CI) | Slope (95% CI) | Mean Difference (ng/mL) | Percent difference mean  |
| --- | --- | --- | --- | --- | --- |
|  223 | 0.9846 | -0.0025 (-0.0195, 0.0145) | 1.0103 (0.9601, 1.0605) | -0.232 | 6.8%  |

Table 20: Bias at Clinical Decision Points

|  Clinical Decision Point (ng/mL) | Bias | Lower CI | Upper CI  |
| --- | --- | --- | --- |
|  0.100 | -0.0022 | -0.0167 | 0.0122  |
|  0.250 | -0.0014 | -0.0151 | 0.0124  |
|  0.500 | 0.0001 | -0.0198 | 0.0199  |
|  2.000 | 0.0087 | -0.0800 | 0.0975  |

Data was analyzed for concordance at the 0.500 ng/mL cut-off. (See Table 21 for 2 x 2 summary.) The percent agreement between the Lumipulse G B•R•A•H•M•S PCT and B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off 0.500 ng/mL for the 223-individual lithium heparin plasma samples tested was determined:

- NPA: 95.5% (84/88), 95% CI: 88.9%, 98.2%
- PPA: 97.0% (131/135), 95% CI: 92.6%, 98.8%

Table 21: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off 0.500 ng/mL

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |
| --- | --- | --- | --- |
|   |  ≤0.500 | >0.500 | Total  |
|  ≤0.500 | 84 | 4 | 88  |
|  >0.500 | 4 | 131 | 135  |
|  Total | 88 | 135 | 223  |

Data was analyzed for concordance at the 2.000 ng/mL cut-off. The percent agreement between the Lumipulse G B•R•A•H•M•S PCT and B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off 2.000 ng/mL for the 223-individual lithium heparin plasma samples tested was determined. See Table 22, 23 and 25 for 2 x 2, 3 x 3 summary tables and regression analyses, respectively. Additionally, Table 24 presents a cross-tabulation of all samples within each medical decision point range

- NPA: 99.2% (132/133), 95% CI: 95.9%, 99.9%
- PPA: 96.7% (87/90), 95% CI: 90.7%, 98.9%

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Table 22: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR at cut-off 2.000 ng/mL

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |
| --- | --- | --- | --- |
|   |  ≤2.000 | >2.000 | Total  |
|  ≤2.000 | 132 | 1 | 133  |
|  >2.000 | 3 | 87 | 90  |
|  Total | 135 | 88 | 223  |

Table 23: 3x3 Table: Lumipulse G B•R•A•H•M•S PCT vs. B•R•A•H•M•S PCT sensitive KRYPTOR

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G B•R•A•H•M•S PCT(ng/mL)  |   |   |   |
| --- | --- | --- | --- | --- |
|   |  ≤2.000 | 0.501<PCT≤2.000 | >2.000 | Total  |
|  ≤2.000 | 84 | 4 | 0 | 88  |
|  0.501<PCT≤2.000 | 4 | 40 | 1 | 45  |
|  >2.000 | 0 | 3 | 87 | 90  |
|  Total | 88 | 47 | 88 | 223  |

Table 24: Cross Tabulation of all Samples (n=223) within each Medical Decision Point Range

|  B•R•A•H•M•S PCT sensitive KRYPTOR (ng/mL) | Lumipulse G PCT (ng/mL)  |   |   |   |   |
| --- | --- | --- | --- | --- | --- |
|   |  0.0237-0.100 | 0.101-0.250 | 0.251-0.500 | 0.501-2.000 | 2.001-58.200  |
|  0.0237-0.100 | 4 | 5 | 0 | 0 | 0  |
|  0.101-0.250 | 6 | 32 | 5 | 2 | 0  |
|  0.251-0.500 | 0 | 6 | 26 | 2 | 0  |
|  0.501-2.000 | 0 | 0 | 4 | 40 | 1  |
|  2.001-58.200 | 0 | 0 | 0 | 3 | 87  |

Table 25: Regression Analyses (N=223)

|   | Parameter | Estimate | Lower CI | Upper CI  |
| --- | --- | --- | --- | --- |
|  Weighted Deming | Intercept | -0.0025 | -0.0195 | 0.0145  |
|   |  Slope | 1.0103 | 0.9601 | 1.0605  |
|  Passing-Bablok | Intercept | -0.005 | -0.017 | 0.006  |
|   |  Slope | 0.995 | 0.977 | 1.013  |

# b. Matrix comparison (Specimen Tube Type Study):

The Lumipulse G B•R•A•H•M•S PCT matrix comparison study was performed to evaluate the difference across tube types (SST, K $_2$ EDTA, Lithium Heparin, Sodium Heparin, and Sodium Citrate) versus the means of the control samples (Red top serum) analyzed per CLSI guideline EP9-A3. Donor blood was drawn into 78 matched sets of collection tubes. The slope for each tube type when compared to the control had  $95\%$  confidence intervals that lay entirely within the range 0.9 to 1.1 and the correlation coefficients were  $\geq 0.9$ . Results are summarized in Table 26 below.

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Table 26: Weighted Deming Regression Analysis Summary of Different Tube Type versus Red Top Tubes

|  Tube Type | n | Concentration Range (ng/mL) |   | Slope |   |   | Intercept |   |   | Pearson Correlation Coefficient  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  Min | Max | Estimate | Lower 95%CI | Upper 95%CI | Estimate | Lower95% CI | Upper95% CI  |   |
|  SST | 77 | 0.014 | 83.435 | 1.0023 | 0.9800 | 1.0246 | -0.0081 | -0.0191 | 0.0029 | 0.9968  |
|  K₂EDTA | 77 | 0.015 | 77.148 | 1.0208 | 1.0002 | 1.0414 | -0.0077 | -0.0190 | 0.0036 | 0.9973  |
|  Li Heparin | 77 | 0.014 | 83.076 | 1.0010 | 0.9560 | 1.0459 | 0.0018 | -0.0121 | 0.0086 | 0.9961  |
|  Na Heparin | 77 | 0.015 | 79.150 | 1.0264 | 1.0070 | 1.0458 | -0.0049 | -0.0200 | 0.0101 | 0.9983  |
|  Citrate | 77 | 0.013 | 87.614 | 1.0399 | 1.0171 | 1.0626 | -0.0095 | -0.0218 | 0.0028 | 0.9963  |

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

b. Clinical specificity:

Not applicable

c. Other clinical supportive data (when a. and b. are not applicable):

Not applicable

4. Clinical cut-offs:

28-day mortality:

ΔPCT ≤ 80%

- A decrease in the PCT levels below or equal to 80% defines a positive ΔPCT test result representing a higher risk for 28-day all-cause mortality of patients diagnosed with severe sepsis or septic shock.

ΔPCT &gt; 80%

- A decrease in the PCT levels of more than 80% defines a negative ΔPCT test result representing a lower risk for 28-day all-cause mortality of patients diagnosed with severe sepsis or septic shock.

Progression Risk:

- PCT &gt; 2 ng/mL: A PCT level above 2.0 ng/mL on the first day of ICU admission is associated with a high risk for progression to severe sepsis and/or septic shock.
- PCT &lt; 0.5 ng/mL: A PCT level below 0.5 ng/mL on the first day of ICU admission is associated with a low risk for progression to severe sepsis and/or septic shock.

LRTI Antibiotic Decision Making:

- PCT &lt; 0.10 ng/mL: Antibiotic therapy strongly discouraged.

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- PCT 0.10-0.25 ng/mL: Antibiotic therapy discouraged.
- PCT 0.26-0.50 ng/mL: Antibiotic therapy encouraged.
- PCT &gt;0.50 ng/mL: Antibiotic therapy strongly encouraged.
- PCT ≤ 0.25 ng/mL: Antibiotic therapy may be discontinued
- ΔPCT &gt; 80%: Antibiotic therapy may be discontinued

Sepsis Antibiotic Discontinuation:

- ΔPCT &gt; 80%: Antibiotic therapy may be discontinued
- PCT ≤ 0.50 ng/mL: Antibiotic therapy may be discontinued

The LUMIPULSE PCT Change Calculator, for use with calculations for assessing Cumulative 28-day Risk of All-Cause Mortality, Initiation and Discontinuation, is a web-based software that will be published on the sponsor's controlled website and made available to end users via the Internet by entering the following URL (www.fujirebio-pct-calculator.com). The calculator software is validated for use on Microsoft Internet Explorer, Google Chrome, Mozilla Firefox, Apple Safari.

## 5. Expected values/Reference range:

Results from a total of 213 samples were used to determine the distribution of Lumipulse G B•R•A•H•M•S PCT results in apparently healthy individuals. Subject demographics for age are presented in Table 27 and subject demographics for race are presented in Table 28 for the apparently healthy subjects. Descriptive statistics for the Lumipulse G B•R•A•H•M•S PCT results are presented in Table 29 for the apparently healthy subjects.

Table 27: Summary of Age Demographics for Healthy Subjects

|  Age Group (years) | Apparently Healthy (All) | Apparently Healthy (Females) | Apparently Healthy (Males)  |
| --- | --- | --- | --- |
|  22-29 | 73 | 38 | 35  |
|  30-39 | 57 | 25 | 32  |
|  40-49 | 57 | 26 | 31  |
|  .50-59 | 21 | 9 | 12  |
|  60-69 | 5 | 3 | 2  |
|  ≥ 70 | 0 | 0 | 0  |
|  Unknown | 0 | 0 | 0  |
|  Total | 213 | 101 | 112  |
|  Mean Age (years) | 7 | 36 | 37  |
|  Median Age (years) | 35 | 33 | 36  |
|  Standard Deviation (SD) (years) | 11 | 11 | 11  |
|  Minimum Age (years) | 22 | 22 | 22  |
|  Maximum Age (years) | 68 | 62 | 68  |

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Table 28: Summary of Race and Ethnicity Demographics for Healthy Subjects

|   | Apparently Healthy (All) | Apparently Healthy (Females) | Apparently Healthy (Males)  |
| --- | --- | --- | --- |
|  White | 87 | 45 | 42  |
|  Black/African American | 40 | 18 | 22  |
|  Hispanic | 61 | 31 | 30  |
|  Asian/Pacific Islander | 0 | 0 | 0  |
|  Other | 0 | 0 | 0  |
|  Unknown | 25 | 7 | 18  |
|  Total | 213 | 101 | 112  |

To determine the upper limit of normal for the Lumipulse G B•R•A•H•M•S PCT assay, calculations of the  $95\%$  reference interval were performed for the overall set of apparently healthy females, apparently healthy males, and combined apparently healthy females and males in the study according to CLSI EP28-A3. These results are presented in Table 29 below.

Table 29: Lumipulse G B•R•A•H•M•S PCT Summary of  $95\%$  Reference Interval for Apparently Healthy Females, Apparently Healthy Males and Combined Apparently Healthy Females and Males

|   | Cohort  |   |   |
| --- | --- | --- | --- |
|   |  Apparently Healthy (All) | Apparently Healthy (Females) | Apparently Healthy (Males)  |
|  N | 213 | 101 | 112  |
|  95% Reference Interval (ng/mL) | 0.003 - 0.045 | 0.003 - 0.029 | 0.004 - 0.045  |
|  Lower 95% Reference Limit (90% Confidence Interval [CI]) (ng/mL) | 0.003 (0.002 - 0.004) | 0.003 (0.002 - 0.004) | 0.004 (0.002 - 0.005)  |
|  Upper 95% Reference Limit (90% CI) (ng/mL) | 0.045 (0.030 - 0.068) | 0.029 (0.017 - 0.065) | 0.045 (0.037 - 0.103)  |

Overall,  $98.1\%$  (209/213) of the apparently healthy subjects had a Lumipulse G B•R•A•H•M•S PCT value equal to or below 0.045 ng/mL. In a population of 213 self-reported healthy individuals, the 95th percentile, upper reference range limit was calculated at 0.045 ng/mL.

It is recommended that each laboratory establish its own range, which may be unique to the population it serves depending upon geographical, patient, and environmental factors.

# N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Parts 801 and 809, as applicable and the special controls for this device type under 21 CFR 866.3215.

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O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

24

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**Source:** [https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PRI/K172713](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PRI/K172713)

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