GELSCAN, MODEL 1206
Device Facts
| Record ID | K100307 |
|---|---|
| Device Name | GELSCAN, MODEL 1206 |
| Applicant | Sebia |
| Product Code | CFE · Clinical Chemistry |
| Decision Date | Aug 31, 2010 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1445 |
| Device Class | Class 2 |
Indications for Use
The SEBIA GELSCAN densitometer is designed and intended for the In Vitro Diagnostic densitometric quantification of electrophoretic separation of human serum Isoenzymes for Lactate Dehydrogenase, Creatine Kinase and Alkaline Phosphatase, on SEBIA HYDRAGEL™ agarose gels, among other SEBIA HYDRAGEL™ electrophoretic assays that are be adaptable to the analyzer depending on the reagents used to induce the electrophoretic analysis. Lactate Dehydrogenase Isoenzymes are used in the diagnosis and treatment of liver diseases, such as viral hepatitis, and myocardial infarction. Creatine Kinase Isoenzymes are used in the diagnosis and treatment of myocardial infarction and muscle diseases such as progressive, Duchenne-type muscular dystrophy. Alkaline Phosphatase Isoenzymes are used in the diagnosis and treatment of liver, bone, parathyroid, and intestinal diseases.
Device Story
The SEBIA GELSCAN is a densitometer for scanning and quantifying electrophoretic agarose gels. It uses a bi-dimensional calibrated CCD sensor to acquire images of gels processed by the SEBIA HYDRASYS system. The device connects to a PC running PHORESIS software, which performs automatic track identification and relative quantification of isoenzyme fractions. It is intended for professional clinical laboratory use. By providing densitometric quantification of electrophoretic patterns, the device assists clinicians in diagnosing and monitoring conditions such as myocardial infarction, liver disease, and muscular dystrophy. The system replaces manual or older densitometric methods with automated digital scanning, improving throughput and consistency in clinical chemistry workflows.
Clinical Evidence
No clinical trials were performed. Evidence consists of analytical performance data, including within-gel and between-scan reproducibility (CVs generally <10%) and method comparison studies. Comparison of 104 LDH, 119 CK, and 48 ALP samples between the GELSCAN and the predicate HYRYS system showed high concordance, with correlation coefficients ranging from 0.843 to 0.999 across various isoenzyme fractions.
Technological Characteristics
Densitometer utilizing a bi-dimensional calibrated CCD sensor for image acquisition. Connects to a PC via wired interface. Dimensions/weight: 5.9 kg. Power: 100-240 V, 50/60 Hz. Software-based quantification of electrophoretic fractions. Designed for use with SEBIA HYDRAGEL agarose gels and HYDRASYS electrophoretic systems.
Indications for Use
Indicated for in vitro diagnostic densitometric quantification of human serum LDH, CK, and ALP isoenzymes separated via electrophoresis on agarose gels. Used to assist in the diagnosis and treatment of liver, bone, parathyroid, intestinal, and muscle diseases, including myocardial infarction and muscular dystrophy.
Regulatory Classification
Identification
A lactate dehydrogenase isoenzymes test system is a device intended to measure the activity of lactate dehydrogenase isoenzymes (a group of enzymes with similar biological activity) in serum. Measurements of lactate dehydrogenase isoenzymes are used in the diagnosis and treatment of liver diseases, such as viral hepatitis, and myocardial infarction.
Special Controls
*Classification.* Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9.
Predicate Devices
- SEBAI HYRYS Densitometer (k960029)
- SEBAI HYDRAGEL ISO-LDH kit (k970477)
- SEBAI HYDRAGEL ISO-CK kit (k992148)
- SEBAI HYDRAGEL ISO-PAL kit (k011113)
Submission Summary (Full Text)
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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY AND INSTRUMENT COMBINATION TEMPLATE
A. 510(k) Number:
k100307
B. Purpose for Submission:
New densitometer
C. Measurand:
LDH isoenzymes, ALP isoenzymes and CPK isoenzymes
D. Type of Test:
Electrophoresis
E. Applicant:
SEBIA
F. Proprietary and Established Names:
GELSCAN, PN 1206
G. Regulatory Information:
1. Regulation section:
21CFR Sec.- 862.1445: Lactate dehydrogenase isoenzymes test system.
21CFR Sec.- 862.1050: Alkaline phosphatase or isoenzymes test system.
21CFR Sec.- 862.1215: Creatine phosphokinase/creatine kinase or isoenzymes test system.
21CFR Sec.- 862.2400: Densitometer/scanner (integrating, reflectance, TLC, or radiochromatogram) for clinical use.
2. Classification:
Class II, II, II, and I respectively
3. Product code:
CFE - Electrophoretic, Lactate Dehydrogenase Isoenzymes
CIN - Electrophoretic Separation, Alkaline Phosphatase Isoenzymes
JHY - Colorimetric Method, CPK or Isoenzymes
JQT - Densitometer/Scanner (Integrating, Reflectance, TLC, Radiochromat.)
Clinical
4. Panel:
75 Chemistry
H. Intended Use:
1. Intended use(s):
See indictment(s) for use below
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2. Indication(s) for use:
The SEBIA GELSCAN densitometer is designed and intended for the In Vitro Diagnostic densitometric quantification of electrophoretic separation of human serum Isoenzymes for Lactate Dehydrogenase, Creatine Kinase and Alkaline Phosphatase, on SEBIA HYDRAGEL™ agarose gels, among other SEBIA HYDRAGEL™ electrophoretic assays that are be adaptable to the analyzer depending on the reagents used to induce the electrophoretic analysis.
Lactate Dehydrogenase Isoenzymes are used in the diagnosis and treatment of liver diseases, such as viral hepatitis, and myocardial infarction.
Creatine Kinase Isoenzymes are used in the diagnosis and treatment of myocardial infarction and muscle diseases such as progressive, Duchenne-type muscular dystrophy.
Alkaline Phosphatase Isoenzymes are used in the diagnosis and treatment of liver, bone, parathyroid, and intestinal diseases.
3. Special conditions for use statement(s):
Prescription use
4. Special instrument requirements:
SEBIA GELSCAN densitometer and the SEBIA HYDRASYS (k960029) electrophoretic system
I. Device Description:
The SEBIA GELSCAN densitometer is an instrument which allows automatic scanning and storage of the HYDRAGEL electrophoresis agarose gels. Data acquisition is performed by a bi-dimensional calibrated CCD sensor. The instrument has to be connected to a PC and the PHORESIS software allows the operator to display the image of the electrophoretic migration by scanning the gel by transmission. The PHORESIS software includes automatic identification of the tracks and automatic relative quantification of the different fractions depending on the assay.
The previously cleared ISO- (LDH, CK and PAL) kits for use on SEBIA GELSCAN densitometer are composed of agarose gels, strips, buffered solvents, lectin, activation solutions, substrates, chromogens, blocking solutions, applicators and filter papers and plastic masks, as necessary for the quantitative electrophoretic analysis of LDH, CPK and ALP isoenzymes
J. Substantial Equivalence Information:
1. Predicate device name(s):
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SEBAI HYRYS Densitometer
SEBAI HYDRAGEL ISO-LDH kit
SEBAI HYDRAGEL ISO-CK kit
SEBAI HYDRAGEL ISO-PAL kit
2. Predicate 510(k) number(s):
k960029, k970477, k992148 and k011113 respectively
3. Comparison with predicate:
| Device | Predicate (k960029) SEBAI HYRYS densitometer | Candidate Device SEBAI GELSCAN densitometer |
| --- | --- | --- |
| Intended Use | The SEBAI HYRYS densitometer is designed and intended for the densitometric quantification of electrophoretic separations of human serum proteins, isoenzymes, lipoproteins and lipoprotein-cholesterol fractions, or human hemoglobins on SEBIA HYDRAGEL agarose gels. The HYDRAGEL kits are used in conjunction with the semi automated HYDRASYS instruments. The electrophoregrams are evaluated visually for pattern abnormalities. Densitometry provides relative quantification of individual zones, | The SEBAI GELSCAN densitometer is designed and intended for the densitometric quantification of electrophoretic separations of human serum isoenzymes (LD, CK and AP), on SEBAI HYDRAGEL agarose gels. The HYDRAGEL kits are used in conjunction with the semi automated HYDRASYS instruments. The electrophoregrams are evaluated visually for pattern abnormalities. Densitometry provides relative quantification of individual zones. |
| Software | HYRYS | PHORESIS |
| Separation System before scanning/densitometry | Agarose gel electrophoresis with the HYDRASYS systems | Same |
| Number of separation units | According to the procedure:
1 agarose gel with small format for 7 analyses
1 agarose gel with large format for 6, 12, 15, 30 & 54 analyses | Same |
| Gel processing for samples analysis before scanning/densitometry | Application of samples, electrophoretic migration, visualization of separated tractions according to the procedure | Same |
| Sample type | Serum | Same |
| Analyzed samples per Scanning | 6,7,12,15,30 or 54 samples | Same |
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| Selection of the sample to scan | Yes | Yes |
| --- | --- | --- |
| Accessory to position and hold the gel for scanning | Exchangeable gel carriers (plates with slots specific for each format) | Gel holders for small and large format and adaptor |
| Samples identification | No | No |
| Design -Manual worklists | Accepts manual worklists | Same |
| Integration device | Screen -Keyboard Integrator Printer | Same |
| User interface | Screen -Keyboard | Same |
| Verification system | Test pattern | Same |
| Results | Electrophoretic pattern | Electrophoretic pattern and gel Picture |
| Number of analysis (throughput) | 2 seconds per sample | 0.5 second per sample |
| Densitometry system for zones quantification | Scanning by transmission on opaque or transparent media, halogen source 12V -20 W with a vertical filament, interferential filters 420 nm, 540 nm and 570 nm standard | Scanning by a bi-dimensional calibrated CCD sensor (Charge-Coupled Device) |
| Human factors | For in vitro diagnostic use. For professional use only | Same |
| Power supply | 115-230 V, 50/60 Hz | 100-240 V, 50160 Hz |
| Electrical, Mechanical and Thermal safety | System certification to EU low voltage safety standards | Same |
| Weight | 20.0 kg | 5.9 kg |
| | ISO-LDH kit k970477 | ISO-LDH kit |
| Densitometer used | SEBAI HYRYS densitometer (k960029) | SEBAI HYRYS densitometer (k960029)and SEBAI GELSCAN densitometer |
| | ISO-CK kit k992148 | ISO-CK kit |
| Densitometer used | SEBAI HYRYS densitometer (k960029) | SEBAI HYRYS densitometer (k960029)and SEBAI GELSCAN densitometer |
| | ISO-PAL kit k011113 | ISO-PAL kit |
| Densitometer used | SEBAI HYRYS densitometer (k960029) | SEBAI HYRYS densitometer (k960029)and SEBAI GELSCAN densitometer |
K. Standard/Guidance Document Referenced (if applicable):
None were referenced.
L. Test Principle:
SEBAI GELSCAN densitometer
Densitometer scanning of stained electrophoregrams yields relative concentrations (percentages) of individual zones. After electrophoresis and visualization, the different fractions are identified according to their position.
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# ISO-LDH kit
Each LD isoenzyme is a tetramer made up of four subunits (polypeptide chains). There are two types of these subunits designated M ("muscle") and H ("heart"). The five LD isoenzymes consist of the five possible combinations of M and H subunits which confer distinct electrophoretic and other properties on each LD isoenzyme. The isoenzymes are designated by their electrophoretic mobility whereby LD1 has been given to the isoenzyme with greatest anodic mobility. The following table summarizes the nomenclature, composition and the primary tissue source of LD isoenzymes:
| NOMENCLATURE | COMPOSITION | TISSUE SOURCE |
| --- | --- | --- |
| LD1 | H4 | Heart (myocardium) |
| LD2 | H3M | Heart (myocardium) |
| LD3 | H2M2 | variable amounts in many tissues |
| LD4 | HM3 | variable amounts in many tissues |
| LD5 | M4 | skeletal muscle, liver |
Diagnosis of myocardial infarction (MI) represents the major value of LD isoenzyme electrophoresis. In normal serum, LD2 is the most prevalent isoenzyme and the LD1 / LD2 ratio is generally < 1. The concentrations of LD1 and, to a lesser degree of LD2, increase after MI and the LD1 / LD2 ratio becomes > 1 (the so called LD1 / LD2 flip). The total LD concentration increases by a factor of two to three within 12 to 24 hours after MI. The LD activity reaches its maximum after two or three days and remains at a high level during about two weeks after infarction. The LD isoenzyme analysis is generally run in tandem with CK (creatine kinase) isoenzymes and / or other early cardiac markers to confirm or rule out the diagnosis of MI, assess its severity and monitor patient's condition. Because of the rather unique distribution of LD isoenzymes in various tissues, their assay in serum aids in diagnosing tissue damage such as in pulmonary and renal infarctions, and hepatic disease. All LD isoenzymes catalyze the same reversible reaction which is utilized in their visualization.
# ISO-CK kit
Creatine kinase isoenzymes consist of two subunits: M ("muscle") and B ("brain"), assembled in dimers. The three resulting combinations constitute the three isoenzymes: MM, principally located in cardiac and skeletal muscles, MB in cardiac muscle and BB in cerebral tissues. Each subunit has a specific electric charge which confers characteristic mobility to the individual CK isoenzymes. The BB fraction is the most anodic, the MM fraction is the most cathodic and the MB is intermediary. The major value of CK isoenzyme electrophoresis is the confirmation of myocardial infarction (MI). The MB concentration increases 4 to 8 hours after myocardial infarction. All CK isoenzymes catalyze the same reaction that is utilized in their
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visualization. Serum samples are electrophoresed and the separated CK isoenzymes are visualized using a specific chromogenic substrate.
# ISO-PAL kit
The alkaline phosphatase (AP) is a metalloglycoprotein with a monophophoesterase activity. It catalyzes the hydrolysis of monophosphoric esters, which is activated by magnesium ions in alkaline medium.
This enzyme is found in many human tissues, e.g., liver, bone, kidney, intestine and placenta. Its elevation is evident in a variety of hepatic, non-hepatic conditions as well as in normal adolescent growth. The enzymes of alkaline phosphatase are coded by 3 structural genes: 2 genes code respectively for the placental and intestinal isoenzymes. The 3rd gene called "non specific tissue gene" is expressed in a variety of tissues such as bone, the liver and kidney.
The individual AP isoenzymes can normally be separated by electrophoresis according to the charge differences. However, the electrophoretic mobilities of the liver and bone isoenzymes are induced only by glycosylation or post-translational modifications and therefore are quite similar. They must be separated with a special treatment of the sample. Quantitation of the AP isoenzymes helps to identify the tissues responsible for the elevation. Electrophoresis of human serum resolves nearly all alkaline phosphatase isoenzymes except the liver 1 (L1) and bone (B), and the placental 1 (P1) when present, migrate together in a single broad fraction.
From anode to cathode, the migration pattern is L1 + B (+ P1); liver 2 (L2, this isoenzymes is also referred to as macrohepatic or fast liver); intestinal 1, 2 and 3 (I1, I2 and I3). The placental 2 (P2), when present, is located between I1 and I2. Additional fraction can be seen in some highly icteric samples close to the application point. It is a complex of alkaline phosphatase and lipoproteins called lipo- ISO-PAL (or ultra fast, UF, in other procedures).
All AP isoenzymes are isolated to certain degree except the intestinal isoenzymes are devoid of sialic acid. The SEBIA system utilizes the different degree of isolation of L1 and B isoenzymes to separate them. Wheat germ lectin (wheat germ agglutinin, WGA) presents a strong affinity for the sialic acid residues and consequently binds preferentially to the bone isoenzyme that is isolated to the highest degree of all. To perform the test, each sample is applied in duplicate. In the course of migration, one of the sample duplicates passes through lectin deposited anodally from the sample's point of application (the lectin itself shifts only slightly cathodally). The interaction between WGA and the passing AP isoenzymes is a reversible reaction.
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# M. Performance Characteristics (if/when applicable):
# 1. Analytical performance:
a. Precision/Reproducibility:
# Iso-LDH
Within gel reproducibility -Three (3) different serum samples were assayed on HYDRASYS instrument using the HYDRAGEL ISO-LDH 7, 15 and 30 procedures and scanned with the GELSCAN densitometer. Each sample was assayed on all tracks of each gel. The mean, SD and CV $(n = 7$ , 15 or 30 according to the procedure) were calculated for each sample and each gel. The following table shows the values of the 3 tested samples for each lactate dehydrogenase fraction calculated with the 3 procedures performed on the HYDRASYS instrument and after densitometric scanning with the GELSCAN.
| FRACTION | LD1 | LD2 | LD3 | LD4 | LD5 |
| --- | --- | --- | --- | --- | --- |
| Pool of normal serum samples: HYDRAGEL 7 ISO-LDH - 15 ISO-LDH - 30 ISO-LDH | | | | | |
| MEAN (%) | 21.4-20.5-21.3 | 35.1-32.8-33.7 | 22.7-26.0-25.4 | 11.0-12.3-11.1 | 9.8-8.3-8.5 |
| SD | 0.26-0.56-0.93 | 0.33-0.83-0.85 | 0.28-0.41-0.60 | 0.38-0.64-0.86 | 0.56-0.62-0.84 |
| CV (%) | 1.2-2.7-4.4 | 1.0-2.5-2.5 | 1.2-1.6-2.4 | 3.5-5.2-7.8 | 5.7-7.4-9.9 |
| Pathological serum sample No.1: HYDRAGEL 7 ISO-LDH - 15 ISO-LDH - 30 ISO-LDH | | | | | |
| MEAN (%) | 14.2-14.4-13.7 | 23.1-21.4-22.3 | 17.3-18.5-18.3 | 11.4-11.4-11.0 | 34.0-34.3-34.8 |
| SD | 0.16-0.43-0.62 | 0.50-0.81-0.57 | 0.45-0.49-0.50 | 0.34-0.81-0.67 | 0.37-0.51-0.59 |
| CV (%) | 1.1-3.0-4.5 | 2.2-3.8-2.6 | 2.6-2.7-2.7 | 3.0-7.1-6.1 | 1.1-1.5-1.7 |
| Pathological serum sample No.2: HYDRAGEL 7 ISO-LDH - 15 ISO-LDH - 30 ISO-LDH | | | | | |
| MEAN (%) | 30.8-31.7-32.6 | 16.4-16.8-16.9 | 20.9-20.5-19.9 | 11.2-10.9-10.6 | 20.8-20.1-20.0 |
| SD | 0.46-0.34-0.51 | 0.16-0.33-0.31 | 0.17-0.48-0.48 | 0.22-0.28-0.21 | 0.27-0.47-0.35 |
| CV (%) | 1.5-1.1-1.6 | 1.0-2.0-1.8 | 0.8-2.3-2.4 | 1.9-2.6-1.9 | 1.3-2.3-1.8 |
Between scans reproducibility without moving the gel on the GELSCAN gel holder - One control serum has been analyzed in reproducibility using the HYDRAGEL 15 ISO-LDH procedure (with 15 analyses per gel) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and without moving the gel on the GELSCAN gel holder. The means, SD and CV $(n = 6)$ were calculated for each analysis of the control serum, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each LD fraction $(n = 15)$ .
| FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| LD1 | 30.7-32.2 | 0.04-0.26 | 0.1-0.8 | 0.4 |
| LD2 | 15.9-17.1 | 0.04-0.11 | 0.2-0.6 | 0.4 |
| LD3 | 19.8-21.4 | 0.04-0.13 | 0.2-0.6 | 0.4 |
| LD4 | 10.6-11.3 | 0.05-0.11 | 0.4-1.0 | 0.7 |
| LD5 | 19.7-21.0 | 0.04-0.29 | 0.2-1.4 | 0.7 |
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Between scans reproducibility after resetting the gel on the GELSCAN gel holder - One control serum has been analyzed in reproducibility using the HYDRAGEL 15 ISO-LDH procedure (with 15 analyses per gel) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and after resetting of the gel on the GELSCAN gel holder. The means, SD and CV $(n = 6)$ were calculated for each analysis of the control serum, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each LD fraction $(n = 15)$ .
| FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| LD1 | 30.8-32.2 | 0.05-0.24 | 0.2-0.8 | 0.4 |
| LD2 | 16.0-17.1 | 0.04-0.12 | 0.2-0.7 | 0.4 |
| LD3 | 19.9-21.5 | 0.05-0.15 | 0.2-0.7 | 0.4 |
| LD4 | 10.6-11.3 | 0.05-0.13 | 0.4-1.2 | 0.7 |
| LD5 | 19.5-20.9 | 0.05-0.27 | 0.2-1.3 | 0.7 |
# Iso-CK
Reproducibility within gel - Three different serum samples were scanned on GELSCAN instrument using the HYDRAGEL 7 ISO-CK, HYDRAGEL 15 ISO-CK and HYDRAGEL 30 ISO-CK procedures. Each sample was run on all tracks of each gel. The mean, SD and CV $(n = 7, 15$ or 30 according to the gel) were calculated for each sample, each fraction and each gel. The following table shows the values of the 3 tested samples for each creatine kinase fraction calculated with the 3 procedures scanned on the GELSCAN instrument.
| FRACTION | BB | MB | MM |
| --- | --- | --- | --- |
| Serum A: HYDRAGEL 7 ISO-CK - 15 ISO-CK - 30 ISO-CK | | | |
| MEAN (%) | / | 8.8-10.1-10.4 | 91.2-89.9-89.6 |
| SD | / | 0.2-0.3-0.6 | 0.2-0.3-0.6 |
| CV (%) | / | 2.5-2.7-5.7 | 0.2-0.3-0.7 |
| Serum B: HYDRAGEL 7 ISO-CK - 15 ISO-CK - 30 ISO-CK | | | |
| MEAN (%) | / | 11.5-12.2-12.8 | 88.5-87.8-87.2 |
| SD | / | 0.5-0.4-0.5 | 0.5-0.4-0.5 |
| CV | / | 4.4-3.3-3.5 | 0.6-0.5-0.5 |
| Serum C: HYDRAGEL 7 ISO-CK - 15 ISO-CK - 30 ISO-CK | | | |
| MEAN (%) | 24.1-27.1-26.9 | 18.2-18.7-18.9 | 57.7-54.2-54.3 |
| SD | 0.5-0.6-0.9 | 0.3-0.3-0.3 | 0.5-0.5-1.1 |
| CV (%) | 2.1-2.4-3.3 | 1.7-1.6-1.8 | 0.8-0.9-2.1 |
Between scans reproducibility without moving the gel on the GELSCAN gel holder - One serum sample has been analyzed in reproducibility using the HYDRAGEL 15 ISO-CK procedure (with 15 analyses per gel) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and without moving the gel on the GELSCAN gel holder. The means, SD and CV $(n = 6)$ were
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calculated for each analysis of this serum sample, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each studied creatine kinase isoenzyme fraction $(n = 15)$ .
| CK FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| BB | 26.2-28.3 | 0.1-0.3 | 0.5-1.2 | 0.8 |
| MB | 18.1-19.1 | 0.1-0.3 | 0.6-1.3 | 1.0 |
| MM | 52.7-55.2 | 0.1-0.4 | 0.3-0.7 | 0.7 |
Between scans reproducibility after resetting the gel on the GELSCAN gel holder - One serum sample has been analyzed in reproducibility using the HYDRAGEL 15 ISO-CK procedure (with 15 analyses per gel) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and after resetting of the gel on the GELSCAN gel holder. The means, SD and CV $(n = 6)$ were calculated for each analysis of this serum sample, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each studied creatine kinase isoenzyme fraction $(n = 15)$ .
| CK FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| BB | 26.2-28.3 | 0.1-0.5 | 0.5-1.6 | 0.9 |
| MB | 18.0-19.1 | 0.1-0.3 | 0.4-1.7 | 0.9 |
| MM | 52.9-55.0 | 0.1-0.5 | 0.2-1.0 | 0.5 |
# ISO PAL
Within gel reproducibility - Three (3) different serum samples were analyzed on HYDRASYS instrument using the HYDRAGEL 7 and 15 ISO-PAL procedures and scanned with the GELSCAN densitometer. Each sample was run on all tracks of each gel. The mean, SD and CV $(n = 3$ or 7 according to the procedure) were calculated for each sample and each gel. The following table shows the values of the 3 tested samples for each studied alkaline phosphatase isoenzyme fraction calculated with the 2 procedures performed on the HYDRASYS instrument and after densitometric scanning with the GELSCAN.
| FRACTION | L1 | L2 | I1 | I2 | I3 | B |
| --- | --- | --- | --- | --- | --- | --- |
| Serum sample No.1: HYDRAGEL 7 - 15 ISO-PAL | | | | | | |
| MEAN (%) | 48.6 -50.2 | 24.0 -24.9 | 1.7 -1.8 | / | / | 25.7 -23.0 |
| SD | 1.03 -1.33 | 0.22 -0.46 | 0.08 -0.18 | / | / | 1.03 -1.11 |
| CV (%) | 2.1 -2.6 | 0.9 -1.8 | 4.8 -9.6 | / | / | 4.0 -4.8 |
| Serum sample No.2: HYDRAGEL 7 - 15 ISO-PAL | | | | | | |
| MEAN (%) | 28.7 -35.2 | 4.7 -4.8 | 15.4 -15.5 | 14.2 -13.9 | 3.7 -3.9 | 33.3 -26.7 |
| SD | 2.29 -2.96 | 0.33 -0.26 | 0.33 -0.62 | 0.46 -0.98 | 0.09 -0.35 | 1.79 -3.13 |
| CV (%) | 8.0 -8.4 | 6.9 -5.5 | 2.1 -4.0 | 3.3 -7.0 | 2.5 -8.9 | 5.4 -11.7 |
| Serum sample No.3: HYDRAGEL 7 - 15 ISO-PAL | | | | | | |
| MEAN (%) | 55.5 -52.6 | 16.1 -16.8 | 6.7 -6.9 | 1.6 -1.7 | 1.2 -0.9 | 18.9 -21.2 |
| SD | 0.59 -1.99 | 0.31 -0.45 | 0.17 -0.50 | 0.16 -0.13 | 0.16 -0.12 | 1.19 -1.65 |
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Between scans reproducibility without moving the gel on the GELSCAN gel holder - One serum sample has been analyzed in reproducibility using the HYDRAGEL 15 ISO-PAL procedure (with 14 analyses per gel, with and without lectin) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and without moving the gel on the GELSCAN gel holder. The means, SD and CV (n = 6) were calculated for each analysis of this serum sample, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each studied alkaline phosphatase isoenzyme fraction (n = 7).
| FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| L1 | 47.2 -51.9 | 0.29 -0.81 | 0.6 -1.6 | 0.9 |
| L2 | 23.8 -25.0 | 0.11 -0.21 | 0.4 -0.9 | 0.6 |
| I1 | 1.6 -2.0 | 0.07 -0.14 | 4.2 -7.4 | 5.5 |
| B | 21.6 -26.0 | 0.28 -0.69 | 1.2 -3.1 | 1.9 |
Between scans reproducibility after resetting the gel on the GELSCAN gel holder - One serum sample has been analyzed in reproducibility using the HYDRAGEL 15 ISO-PAL procedure (with 14 analyses per gel, with and without lectin) on the HYDRASYS instrument with 6 successive densitometric scannings on the GELSCAN and after resetting of the gel on the GELSCAN gel holder. The means, SD and CV (n = 6) were calculated for each analysis of this serum sample, each zone and on the 6 scans. The table shows the limit values for the tested sample and a mean CV calculated from the CV's for each studied alkaline phosphatase isoenzyme fraction (n = 7).
| FRACTION | MEAN (%) | SD | CV (%) | MEAN CV (%) |
| --- | --- | --- | --- | --- |
| L1 | 46.9 -51.5 | 0.09 -0.42 | 0.2 -0.9 | 0.5 |
| L2 | 24.0 -25.3 | 0.04 -0.15 | 0.2 -0.6 | 0.3 |
| I1 | 1.6 -1.9 | 0.05 -0.09 | 2.7 -6.0 | 3.8 |
| B | 22.0 -26.4 | 0.09 -0.47 | 0.4 -2.0 | 1.0 |
b. Linearity/assay reportable range:
Electophoretic assays report percentages of analyte fractions. Various dilutions of each analyte spanning ranges the of total activity; 750 – 50 IU/L for LDH, 750 – 20 IU/L for CK and 1000 – 10 IU/L for ALP show approximately equal percentages per fraction for each series of dilutions with CVs ranging from 1.3 to 11%.
c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability was not provided.
d. Detection limit:
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Each enzyme was diluted to various levels and electrophoresed to determine the lowest detectable level of isoenzymes. The lowest detected activity of the individual isoenzymes by densitometry was 2 IU/L for all LD fractions, 2.5 IU/L for the CKMB fraction and ranged from 1 to 4 IU/L for all ALP fractions.
e. Analytical specificity: Not Applicable
f. Assay cut-off: Not Applicable
# 2. Comparison studies:
a. Method comparison with predicate device:
The analysis of 104 different pathological and normal serum samples, by agarose gel electrophoresis using the HYDRAGEL 7, 15 and 30 ISO-LDH procedures with the HYDRASYS system and GELSCAN scanning or HYRYS densitometry, demonstrated concordance between these two quantification systems for the five lactate dehydrogenase fractions. The correlation parameters calculated for individual zones from the pooled data for HYDRAGEL 7, 15 and 30 ISO-LDH and GELSCAN vs. the comparative system (y = HYDRAGEL 7, 15 and 30 ISO-LDH and GELSCAN) were :
| LD Fraction | Correlation coefficient | y-intercept | Slope | Range of % values HYDRAGEL 7,15 & 30 ISO-LDH (GELSCAN) |
| --- | --- | --- | --- | --- |
| LD1 | 0.984 | 1.972 | 0.947 | 7.7-33.4 |
| LD2 | 0.983 | 2.216 | 0.891 | 13.6-44.7 |
| LD3 | 0.972 | 2.723 | 0.859 | 14.4-29.9 |
| LD4 | 0.843 | 0.869 | 0.952 | 6.4-19.7 |
| LD5 | 0.989 | 1.059 | 0.972 | 5.1-52.9 |
The analysis of 119 different pathological and normal serum samples, by agarose gel electrophoresis using the HYDRAGEL 7, 15 and 30 ISO-CK procedures with the HYDRASYS system and GELSCAN scanning or HYRYS densitometry, demonstrated concordance between these two quantification systems for the different BB, MB and MM creatine kinase isoenzymes fractions. The correlation parameters calculated for individual zones from the pooled data for HYDRAGEL 7, 15 and 30 ISO-CK & GELSCAN vs. the comparative system (y = HYDRAGEL 7, 15 and 30 ISO-CK & GELSCAN) were :
| CK Fraction | Correlation coefficient | y-intercept | Slope | Range of % values HYDRA GEL ISO-CK (GELSCAN) |
| --- | --- | --- | --- | --- |
| BB | 0.997 | -0.552 | 1.036 | 1.0-25.8 |
| MB | 0.997 | -0.552 | 1.036 | 1.0-25.8 |
| MM | 0.997 | -0.552 | 1.036 | 1.0-25.8 |
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The analysis of 48 different pathological and normal serum samples, by agarose gel electrophoresis using the HYDRAGEL 7 and 15 ISO-PAL procedures with the HYDRASYS system and GELSCAN scanning or HYRYS densitometry, demonstrated concordance between these two quantification systems for the different L1 + P1, L2, I1, P2, I2, I3 and Bone alkaline phosphatase isoenzymes fractions. The correlation parameters calculated for individual zones from the pooled data for HYDRAGEL 7 and 15 ISO-PAL & GELSCAN vs. the comparative system (y = HYDRAGEL 7 and 15 ISO-PAL & GELSCAN) were :
| ALP Fraction | Correlation coefficient | y-intercept | Slope | Range of % values HYDRAGEL 7 & 15 ISO-PAL(GELSCAN) |
| --- | --- | --- | --- | --- |
| L1 + P1 | 0.994 | 1.356 | 0.993 | 4.2 -76.9 |
| L2 | 0.998 | -0.033 | 1.002 | 0.0 -71.2 |
| I1 | 0.999 | 0.013 | 0.985 | 0.0 -38.7 |
| P2 | 0.999 | 0.013 | 0.981 | 0.0 -18.6 |
| I2 | 0.999 | 0.027 | 1.003 | 0.0 -19.4 |
| I3 | 0.998 | 0.022 | 0.875 | 0.0 -3.6 |
| B | 0.990 | 1.069 | 0.941 | 6.2 -89.0 |
b. Matrix comparison: Not applicable
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-off: Not applicable
5. Expected values/Reference range: Same as in the cleared device for k970477, k992148 and k011113
N. Instrument Name:
SEBIA GELSCAN densitometer
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O. System Descriptions:
1. Modes of Operation:
Does the applicant’s device contain the ability to transmit data to a computer, webserver, or mobile device?:
Yes ☐ X or No ☐
Does the applicant’s device transmit data to a computer, webserver, or mobile device using wireless transmission?:
Yes ☐ or No ☐ X
2. Software:
FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:
Yes ☐ X or No ☐
3. Specimen Identification:
Worklist generated by data entered
4. Specimen Sampling and Handling:
Manually
5. Calibration:
Camera calibration
6. Quality Control:
Contains Quality Control software application
P. Other Supportive Instrument Performance Characteristics Data Not Covered In The “Performance Characteristics” Section above:
None
Q. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
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R. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
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