Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard in vitro diagnostic assay for measuring bilirubin levels using chemical reagents and a clinical chemistry analyzer. There is no mention of AI/ML algorithms, image processing, or data training/testing sets typically associated with AI/ML-powered devices.

Therapeutic

No
This device is an in vitro diagnostic test for measuring direct bilirubin levels, used in the diagnosis and treatment of various disorders, but it does not directly treat or prevent a disease.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the measurement of bilirubin levels using this device "is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block." This directly indicates a diagnostic purpose.

SaMD (Software as a Medical Device)

No

The device is a reagent kit for an in vitro diagnostic test, which includes chemical components (reagents) and is used on a hardware system (COBAS INTEGRA systems). It is not solely software.

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 that the device is an "in vitro test for the quantitative determination of direct bilirubin in human serum and plasma". This clearly indicates that the device is intended for use outside of the body to analyze biological samples.
Purpose: The intended use also describes the purpose of the test: "Measurement of the levels of bilirubin... is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders...". This aligns with the definition of an IVD, which is used to provide information for the diagnosis, treatment, or prevention of disease.
Sample Type: The device analyzes "human serum and plasma", which are biological specimens.
Device Description: The description details the reagents used to perform the test on these biological samples.

All of these points are consistent with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

COBAS INTEGRA Bilirubin Direct Gen.2 is an in vitro test for the quantitative determination of direct bilirubin in human serum and plasma on COBAS INTEGRA systems. Measurement of the levels of bilirubin, an organic compound formed during the normal and abnormal destruction of red blood cells, is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block.

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

CIG

Device Description

COBAS INTEGRA Bilirubin Direct Gen.2 reagent provides quantitative measurement of the direct bilirubin that is present in a human serum or human plasma sample. Reagents are packaged in a cassette with two bottles labeled with their instrument positioning, R1 and SR. R1, or Reagent 1, contains Phosphoric acid 85 mmol/L, NaCl 50 mmol/L, and HEDTA 4.0 mmol/L at pH 1.9. SR, or Start Reagent, is a 3,5-dichlorophenyl diazonium salt at 1.5 mmol/L in acid buffer, pH 1.3.

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

Not Found

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)

Precision/reproducibility: Precision was determined according to CLSI EP5-A2. The study included human sera samples (0.12, 3.76, and 13.2 mg/dL) and two serum-based control samples in two aliquots per run and two runs per day for 21 days.
Linearity/ assay reportable range: Linearity was assessed according to CLSI EP6-A with one batch of reagent, in one run, and with samples measured in triplicate. Two separate dilution series differing by sample type (serum and plasma) were prepared with thirteen levels each. Lithium-heparin was used to prepare the plasma sample series. The highest concentration samples exceed the desired measuring range. The highest concentration samples were created by taking low analyte native samples and spiking them with ditaurobilirubin.
Detection limit: LoB. LoD, and LoQ studies were performed based upon CLSI EP17-A2.
Method comparison with predicate device: Direct bilirubin values for n=71 human sera samples were obtained using the candidate reagent (y-axis) to the predicate reagent (x-axis) on the COBAS INTEGRA 800 clinical chemistry analyzer. Samples ranged from 0.083 to 13.762 mg/dL and were tested in singlicate. The values were regressed using the Passing/Bablok model to produce the following equation. R2 = 0.9979 y = 1.0490x + 0.0699 mg/dL
Matrix comparison: Lithium-heparin, K2-EDTA, and K3-EDTA are permissible anticoagulants for use with this reagent because they do not interfere with recovery of direct bilirubin. 32 tubes were collected per anticoagulant. Plasma results were compared to serum results and percent recovery was determined.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Repeatability Summary:
Total Mean (mg/dL) for PNU: 0.75, PPU: 1.9, Human Serum 1: 0.12, Human Serum 2: 3.8, Human Serum 3: 13.2
Within Run Imprecision SD (mg/dL): PNU: 0.01, PPU: 0.01, Human Serum 1: 0.01, Human Serum 2: 0.01, Human Serum 3: 0.04
Within Run Imprecision CV%: PNU: 1.2, PPU: 0.6, Human Serum 1: 7.4, Human Serum 2: 0.4, Human Serum 3: 0.3
Min (mg/dL): PNU: 0.72, PPU: 1.9, Human Serum 1: 0.09, Human Serum 2: 3.7, Human Serum 3: 13.1
Max (mg/dL): PNU: 0.78, PPU: 2.0, Human Serum 1: 0.13, Human Serum 2: 3.8, Human Serum 3: 13.3

Intermediate Precision:
Total Mean (mg/dL) for PNU: 0.75, PPU: 1.9, Human Serum 1: 0.12, Human Serum 2: 3.8, Human Serum 3: 13.2
Total Imprecision SD (mg/dL): PNU: 0.01, PPU: 0.02, Human Serum 1: 0.01, Human Serum 2: 0.04, Human Serum 3: 0.05
Total Imprecision CV%: PNU: 1.6, PPU: 1.0, Human Serum 1: 7.7, Human Serum 2: 1.0, Human Serum 3: 0.4
Min (mg/dL): PNU: 0.72, PPU: 1.9, Human Serum 1: 0.09, Human Serum 2: 3.7, Human Serum 3: 13.1
Max (mg/dL): PNU: 0.78, PPU: 2.0, Human Serum 1: 0.13, Human Serum 2: 3.8, Human Serum 3: 13.3

Measuring Ranges that are Supported by the Linearity Data:
Plasma: Range tested (mg/dL) 0.01 - 19.5, Range found (mg/dL) 0.01 - 19.5, Recommended measuring range (mg/dL) 0.07 - 13.8
Serum: Range tested (mg/dL) 0.02 - 19.4, Range found (mg/dL) 0.02 - 17.4, Recommended measuring range (mg/dL) 0.07 - 13.8
Linear Regression Equation for Serum y = 1.0000x ~ 0.0000 r2 = 0.9944
Linear Regression Equation for Plasma r2 = 0.9977 y = 1.0000x - 0.0000

Detection limit: LoB claim = 0.05 mg/dL, LoD claim = 0.07 mg/dL, LoQ claim = 0.07 mg/dL

Endogenous Interference Summary Data:
Lipemia low analyte: 1098
Lipemia high analyte: 1100
Hemolysis low analyte: 35
Hemolysis high analyte: 25
The lowest L index for which there is no significant interference is 1098. The lowest H index for which there is no significant interference is 25.

Drug interference: "Phenylbutazone causes falsely low bilirubin results." The remaining 17 commonly used drugs produce no interference with BILD2 measurement.

Method comparison with predicate device: y = 1.0490x + 0.0699 mg/dL; R2 = 0.9979

Median Values for Anticoagulant Comparisons:
anticoagulants | median recovery | median absolute deviation(mg/dL)
Li-Heparin (full) | 102% | +0.02
Li-Heparin (half) | 102% | -0.05
K2-EDTA (full) | 101% | +0.00
K2-EDTA (half) | 101% | -0.02
K3-EDTA (full) | 100% | -0.00
K3-EDTA (half) | 100% | -0.03
Gel Separation Tube | 104% | +0.02
Criteria | 90 to 110% |

Regulation Number and Section

§ 862.1110 Bilirubin (total or direct) test system.

(a)
Identification. A bilirubin (total or direct) test system is a device intended to measure the levels of bilirubin (total or direct) in plasma or serum. Measurements of the levels of bilirubin, an organic compound formed during the normal and abnormal distruction of red blood cells, if used in the diagnosis and treatment of liver, hemolytic hematological, and metabolic disorders, including hepatitis and gall bladder block.(b)
Classification. Class II.

Summary

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510(k) Summary for COBAS INTEGRA Bilirubin Direct Gen.2

510(k) number	K123965
Purpose of submission	Roche Diagnostics hereby submits this 510(k) to provide FDA with notification of intent to market a new device named COBAS INTEGRA Bilirubin Direct Gen.2 reagent.
This candidate device is a new reagent that was developed by Roche Diagnostics. The previous generation of reagent, COBAS INTEGRA Bilirubin Direct, was cleared in 510(k) K063543 and serves as the predicate device. The candidate and predicate devices use the same calibrator and controls. Only the reagents differ. This submission presents data to support clearance of this new reagent.
Measurand	Direct Bilirubin
Type of test	Quantitative diazo colorimetric method
Applicant	Roche Diagnostics
Candidate device names	Proprietary name:
COBAS INTEGRA Bilirubin Direct Gen.2
Common name:
Bilirubin Direct Gen.2
Regulatory information	Product CodeClassificationRegulationPanelCIGClass II21 CFR 862.1110 (Bilirubin (total or direct) test system)Clinical Chemistry (75)	Product Code	Classification	Regulation	Panel	CIG	Class II	21 CFR 862.1110 (Bilirubin (total or direct) test system)	Clinical Chemistry (75)
Product Code	Classification	Regulation	Panel
CIG	Class II	21 CFR 862.1110 (Bilirubin (total or direct) test system)	Clinical Chemistry (75)

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| Intended use | In vitro test for the quantitative determination of direct bilirubin in human
serum and plasma on COBAS INTEGRA systems. |
|---------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Indications for
use | COBAS INTEGRA Bilirubin Direct Gen.2 is an in vitro test for the
quantitative determination of direct bilirubin in human serum and plasma on
COBAS INTEGRA systems. Measurement of the levels of bilirubin, an
organic compound formed during the normal and abnormal destruction of red
blood cells, is used in the diagnosis and treatment of liver, hemolytic,
hematological, and metabolic disorders, including hepatitis and gall bladder
block. |
| Special
conditions for
use | For prescription use only |
| Special
instrument
requirements | For use on the COBAS INTEGRA clinical chemistry analyzer |
| Candidate
device
description | COBAS INTEGRA Bilirubin Direct Gen.2 reagent provides quantitative
measurement of the direct bilirubin that is present in a human serum or
human plasma sample.

Reagents are packaged in a cassette with two bottles labeled with their
instrument positioning, R1 and SR. R1, or Reagent 1, contains Phosphoric
acid 85 mmol/L, NaCl 50 mmol/L, and HEDTA 4.0 mmol/L at pH 1.9. SR,
or Start Reagent, is a 3,5-dichlorophenyl diazonium salt at 1.5 mmol/L in acid
buffer, pH 1.3. |
| Predicate
device | Roche Diagnostics claims substantial equivalence to the COBAS INTEGRA
Bilirubin Direct reagent that was cleared with the Special 510(k) K063543. |
| | Continued on next page |

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The following table compares the identical features of the candidate device to Substantial equivalence the predicate device that was cleared in 510(k) K063543. similarities

| Feature | Predicate Device:
Bilirubin Direct | Candidate Device:
Bilirubin Direct Gen.2 |
|-----------------------|----------------------------------------------------------------------------------------------|---------------------------------------------|
| Sample Types | Serum and plasma | Same |
| Reference Method | Diazo colorimetric method | Same |
| Calibrator | Calibrator for automated systems
(C.f.a.s.) and deionized water as the
zero calibrator | Same |
| Calibration Stability | Recalibrate with each lot as and
required following quality control
procedures | Same |
| Calibration Mode | Linear regression | Same |
| Traceability | Standardized against the Doumas
manual reference method | Same |

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The following table compares the different features of the candidate device to Substantial the predicate device that was cleared in 510(k) K063543. equivalence differences .

| Feature | Predicate Device:
Bilirubin Direct | Candidate Device:
Bilirubin Direct Gen.2 |
|---------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | COBAS INTEGRA Bilirubin
Direct (BIL-D) contains an in
vitro diagnostic reagent system
intended for use on COBAS
INTEGRA systems for the
quantitative determination of the
direct (conjugated) bilirubin
concentration in serum and
plasma. | In vitro test for the quantitative
determination of direct bilirubin in
human serum and plasma on
COBAS INTEGRA systems. |
| Indications for Use | The cassette COBAS INTEGRA
Bilirubin Direct (BIL-D) contains
an in vitro diagnostic reagent
system intended for use on
COBAS INTEGRA systems for
the quantitative determination of
the direct (conjugated) bilirubin
concentration in serum and
plasma (test BIL-D, 0-049).
Measurement of the levels of
bilirubin, an organic compound
formed during the normal and
abnormal destruction of red blood
cells, is used in the diagnosis of
liver, hemolytic, hematological,
and metabolic disorders, including
hepatitis and gall bladder block. | COBAS INTEGRA Bilirubin
Direct Gen.2 is an in vitro test for
the quantitative determination of
direct bilirubin in human serum
and plasma on COBAS INTEGRA
systems.
Measurement of the levels of bilirubin, an organic compound formed during the normal and abnormal destruction of red blood cells, is used in the diagnosis and treatment of liver, hemolytic, hematological, and metabolic disorders, including hepatitis and gall bladder block. |
| | Permissible
Anticoagulants | Li-heparin |
| Instrument Platform | COBAS INTEGRA 400,
400 Plus, 700, and 800 | COBAS INTEGRA 800 |

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Substantial equivalence - differences continued

| Feature | Predicate Device:
Bilirubin Direct | Candidate Device:
Bilirubin Direct Gen.2 |
|---------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Reagent Composition | R1:
Sulfanilic acid 35 mmol/L,
Oxalic acid 40 mmol/L,
HEDTA 4.0 mmol/L, and
pH 1.2

R2:
Sodium nitrite 3.9 mmol/L and
pH 6.0

Sulfanilic acid reacts with sodium
nitrite to form diazotized
sulfanilic acid. | R1:
Phosphoric acid 85 mmol/L,
NaCl 50 mmol/L,
HEDTA 4.0 mmol/L, and
pH 1.9

COBAS INTEGRA 400/400 plus:
10-15 °C for 8 weeks | COBAS INTEGRA 800:
8 °C for 6 weeks |
| Controls | Precinorm U plus,
Precipath U plus,
Precinorm U,
Precipath U | Precinorm U plus,
Precipath U plus,
PreciControl ClinChem Multi 1A,
PreciControl ClinChem Multi 2A

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| Test principle | COBAS INTEGRA Bilirubin Direct Gen.2 measures direct bilirubin by
employing the diazo colorimetric method. Conjugated bilirubin and δ-bilirubin (direct bilirubin) react directly with 3,5-dichlorophenyl diazonium
salt in acid buffer to form the red-colored azobilirubin. The color intensity of
the red azobilirubin formed is directly proportional to the direct bilirubin
concentration. The color intensity is measured photometrically by a COBAS
INTEGRA clinical chemistry analyzer. |
|-------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Precision/
reproducibility | Precision was determined according to CLSI EP5-A2. The study included
human sera samples (0.12, 3.76, and 13.2 mg/dL) and two serum-based
control samples in two aliquots per run and two runs per day for 21 days.

Here are summaries of the repeatability and intermediate precision data. |
| Repeatability Summary | |

Specimen	PNU	PPU	Human Serum 1	Human Serum 2	Human Serum 3
Total Mean (mg/dL)	0.75	1.9	0.12	3.8	13.2
Within Run Imprecision
SD (mg/dL)	0.01	0.01	0.01	0.01	0.04
Within Run Imprecision
CV%	1.2	0.6	7.4	0.4	0.3
Min (mg/dL)	0.72	1.9	0.09	3.7	13.1
Max (mg/dL)	0.78	2.0	0.13	3.8	13.3

Intermediate Precision

Specimen	PNU	PPU	Human Serum 1	Human Serum 2	Human Serum 3
Total Mean (mg/dL)	0.75	1.9	0.12	3.8	13.2
Total Imprecision
SD (mg/dL)	0.01	0.02	0.01	0.04	0.05
Total Imprecision
CV%	1.6	1.0	7.7	1.0	0.4
Min (mg/dL)	0.72	1.9	0.09	3.7	13.1
Max (mg/dL)	0.78	2.0	0.13	3.8	13.3

Values that appear in bold type also appear in the labeling.

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Linearity/ assay reportable range

Linearity was assessed according to CLSI EP6-A with one batch of reagent, in one run, and with samples measured in triplicate. Two separate dilution series differing by sample type (serum and plasma) were prepared with thirteen levels each. Lithium-heparin was used to prepare the plasma sample series. The highest concentration samples exceed the desired measuring range. The highest concentration samples were created by taking low analyte native samples and spiking them with ditaurobilirubin.

Measuring Ranges that are Supported by the Linearity Data

	Plasma	Serum
Range tested (mg/dL)	0.01 - 19.5	0.02 - 19.4
Range found (mg/dL)	0.01 - 19.5	0.02 - 17.4
Recommended measuring range (mg/dL)	0.07 - 13.8	0.07 - 13.8

The quadratic model is significant for both sample types.

Linear Regression Equation for Serum y = 1.0000x ~ 0.0000 r2 = 0.9944

Linear Regression Equation for Plasma r2 = 0.9977 y = 1.0000x - 0.0000

Traceability, stability, and expected values

This method has been standardized against the manual test performance using the Doumas method.

The reagent has been evaluated for transport, shelf-life, open on-board, and calibration stability.

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Detection limit LoB. LoD, and LoO studies were performed based upon CLSI EP17-A2.

LoB Protocol: One blank sample was tested in n=5 with two analyzers with three reagent batches for two runs per day across three days.

LoD Protocol: Five low-analyte samples were measured in singlicate on two analyzers with three reagent batches for two runs per day across three days.

LoO Protocol: A low-level sample set of nine was measured in singlicate, using three reagent batches on two analyzers for two runs per day across three days. The LoQ is determined based on precision at 20% CV.

The LoB, LoD, and LoQ claims represent the specifications for each.

LoB claim = 0.05 mg/dL LoD claim = 0.07 mg/dL LoQ claim = 0.07 mg/dL

Analytical specificity interference from endogenous substances

The reagent was evaluated with two endogenous substances, hemoglobin and lipids, for potential interference with the measurement of direct bilirubin.

One pool of human serum was spiked with the interferent. A second pool of human serum contained none. The two pools were mixed in different ratios to yield a dilution series with varying concentrations of the interferent (from 0 to 10).

The endogenous interference data are summarized in the table. The labeling claims the specification, "No significant interference up to an H index of 25," and "No significant interference up to an L index of 750."

Endogenous Interference Summary Data

	no interference up to this concentration (mg/dl)
Lipemia low analyte	1098
Lipemia high analyte	1100
Hemolysis low analyte	35
Hemolysis high analyte	25

The lowest L index for which there is no significant interference is 1098. The lowest H index for which there is no significant interference is 25.

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Analytical specificity interference from common drugs

Eighteen commonly used drugs were added to native patient samples and examined for potential interference on measurement with COBAS INTEGRA Bilirubin Direct reagent.

Drug interference testing was performed with serum sample pools at two target concentrations of direct bilirubin, one at a low concentration of ~ 1.8 mg/dL and the second one at a high concentration of ~ 4.9 mg/dL.

Direct bilirubin concentration in all aliquots is measured in triplicate on the COBAS INTEGRA analyzer. The mean value among the triplicates for each aliquot is determined. From the mean values, the percent recovery to the initial value is calculated.

"Phenylbutazone causes falsely low bilirubin results." This statement appears in the labeling. The remaining 17 commonly used drugs produce no interference with BILD2 measurement.

| | Drug | Highest Concentration Shown
Not to Interfere with BILD2
(mg/L, except Heparin) |
|----|----------------------|--------------------------------------------------------------------------------------|
| 1 | Acetylcystein | 150 |
| 2 | Ampicillin - Na | 1000 |
| 3 | Ascorbic acid | 300 |
| 4 | Ca - Dobesilate | 200 |
| 5 | Cyclosporine A | 5 |
| 6 | Cefoxitin | 2500 |
| 7 | Heparin - Na | 5000 U |
| 8 | Intralipid | 10000 |
| 9 | Levodopa | 20 |
| 10 | Methyldopa + 1.5 | 20 |
| 11 | Metronidazole | 200 |
| 12 | Doxycyclin | 50 |
| 13 | Acetylsalycilic acid | 1000 |
| 14 | Rifampicin | 60 |
| 15 | Acetaminophen | 200 |
| 16 | Ibubrofen | 500 |
| 17 | Theophylline | 100 |

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Method comparison with predicate device

Direct bilirubin values for n=71 human sera samples were obtained using the candidate reagent (y-axis) to the predicate reagent (x-axis) on the COBAS INTEGRA 800 clinical chemistry analyzer. Samples ranged from 0.083 to 13.762 mg/dL and were tested in singlicate. The values were regressed using the Passing/Bablok model to produce the following equation. R2 = 0.9979

y = 1.0490x + 0.0699 mg/dL

Matrix comparison Lithium-heparin, K2-EDTA, and K3-EDTA are permissible anticoagulants for use with this reagent because they do not interfere with recovery of direct bilirubin. 32 tubes were collected per anticoagulant. Plasma results were compared to serum results and percent recovery was determined.

Median Values for Anticoagulant Comparisons

| anticoagulants | median recovery | median absolute
deviation(mg/dL) |
|---------------------|-----------------|-------------------------------------|
| Li-Heparin (full) | 102% | +0.02 |
| Li-Heparin (half) | 102% | -0.05 |
| K2-EDTA (full) | 101% | +0.00 |
| K2-EDTA (half) | 101% | -0.02 |
| K3-EDTA (full) | 100% | -0.00 |
| K3-EDTA (half) | 100% | -0.03 |
| Gel Separation Tube | 104% | +0.02 |
| Criteria | 90 to 110% |