(408 days)
The Neonatal Biotinidase kit is intended for the semiquantitative determination of biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency.
Biotinidase is found in the blood sample itself. Filter paper disks from newborn dried blood spot samples, calibrators and controls are punched into the wells of a microplate. When biotin substrate reagent containing biotin 6-aminoquinoline (6-AQ) is added to a well containing a punched dried blood spot, the reagent extracts and reconstitutes the proteins and enzymes in the spot. The biotinidase enzyme in the sample cleaves the substrate to biotin and fluorescent 6-AQ The addition of the ethanol stops the reaction and precipitates the proteins to cover the bottom of the well and the extracted spot. The fluorescent product (6-AQ) formed during the reaction is measured with a fluorometer. The biotinidase activity is defined against a calibration curve. The biotinidase activity of the sample is determined by comparing the fluorescence intensity of the sample to a calibration curve.
The Neonatal Biotinidase kit is intended for the semi-quantitative determination of biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency. The study aimed to demonstrate substantial equivalence to the Astoria-Pacific SPOTCHECK® Biotinidase 50-Hour Reagent Kit (K010844).
1. Table of Acceptance Criteria and Reported Device Performance:
The document describes a method comparison study to establish substantial equivalence rather than explicit acceptance criteria with pre-defined thresholds. However, based on the provided results, the implicit acceptance criteria would involve high positive percent agreement (PPA) and overall percent agreement (OPA) with the predicate device.
| Metric | Acceptance Criteria (Implicit) | Reported Device Performance |
|---|---|---|
| Positive Percent Agreement (PPA) | High agreement with predicate for positive cases | 90.9% (20/22) |
| Overall Percent Agreement (OPA) | High agreement with predicate for all cases | 99.8% ((20+1493)/1516) |
2. Sample Size Used for the Test Set and Data Provenance:
- Test Set Sample Size: 1516 samples.
- Data Provenance: The samples included 1516 newborn dried blood spot specimens representing the US population. The study included:
- 1496 routine screening specimens (prospective, from the US population).
- 20 retrospective specimens diagnosed positive for biotinidase deficiency (provenance not explicitly stated but implies confirmed cases, likely from a US population or referral lab).
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications:
The document does not specify the number or qualifications of experts used to establish the ground truth. It states that 20 retrospective specimens were "diagnosed positive for biotinidase deficiency," implying a clinical diagnosis, but details on who made these diagnoses are not provided. For the routine screening samples, the ground truth is established by the comparison to the predicate device and clinical outcome (diagnosed biotinidase deficiency or not).
4. Adjudication Method for the Test Set:
Not applicable. This study is a method comparison between two assays, not a diagnostic study requiring adjudication against a clinical outcome based on expert review of ambiguous cases. The "ground truth" for the deficient samples appears to be established by prior clinical diagnosis, and routine samples are compared against the predicate as well as clinical diagnosis outcomes.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:
No, an MRMC comparative effectiveness study was not done. This study is a comparison of two diagnostic assays, not an assessment of human reader performance with or without AI assistance.
6. Standalone (Algorithm Only) Performance Study:
Yes, a standalone study was performed. The device (Neonatal Biotinidase kit) was tested on its own and its results were compared to those from a commercially available predicate device. The performance metrics (PPA, OPA) are based on the direct output of the device.
7. Type of Ground Truth Used:
The ground truth for the 20 positive cases was established by clinical diagnosis ("retrospective specimens diagnosed positive for biotinidase deficiency"). For the routine samples, the "ground truth" for comparison purposes within the study was intrinsically linked to the results of the predicate device and the absence of a biotinidase deficiency diagnosis.
8. Sample Size for the Training Set:
The document does not provide information on a specific training set or its sample size for the Neonatal Biotinidase kit. Kits for in vitro diagnostic (IVD) use typically involve extensive assay development and validation, but specific "training sets" in the machine learning sense are not usually reported in this context. The calibration curve and internal controls are part of the assay's design and standardization.
9. How the Ground Truth for the Training Set Was Established:
As no specific training set in the AI/ML sense is mentioned, information on how its ground truth was established is not provided. The kit's calibration curve is established using six levels of dried blood spots prepared from porcine blood, representing different biotinidase activity levels, which serves as a form of internal reference material.
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510(k) Summary
This summary of 510(k) safety and effectiveness information is supplied in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.
The assigned 510 (k) number is K090123
Kay A. Taylor
Tele: 317 418-1735 Fax: 317 536-3064
Helena Lundstrom
Tele: (011) +358-2-2678575 Fax: (011) +358-2-2678357
Neonatal Biotinidase kit
Neonatal Biotinidase kit
Date: March 5, 2010
Submitted by: Wallac Ov Mustionkatu 6 20750 Turku, Finland
Contact Person:
Primary:
Secondary:
Trade Name:
Common Name:
Classification Name:
System, Test, Biotinidase (21 CFR 862.1118/ Product Code NAK)
Astoria-Pacific SPOTCHECK® Biotinidase 50-Hour Reagent Predicate device: Kit, (K010844)
Biotinidase is found in the blood sample itself. Filter paper Device Description: disks from newborn dried blood spot samples, calibrators and controls are punched into the wells of a microplate. When biotin substrate reagent containing biotin 6-aminoquinoline (6-AQ) is added to a well containing a punched dried blood spot, the reagent extracts and reconstitutes the proteins and enzymes in the spot. The biotinidase enzyme in the sample cleaves the substrate to biotin and fluorescent 6-AQ The addition of the ethanol stops the reaction and precipitates the proteins to cover the bottom of the well and the extracted spot. The fluorescent product (6-AQ) formed during the reaction is measured with a fluorometer. The biotinidase activity is defined against a calibration curve. The biotinidase activity of the sample is
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determined by comparing the fluorescence intensity of the sample to a calibration curve.
Intended Use:
The Neonatal Biotinidase kit is intended for the semiquantitative determination of biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency.
Device Comparison:
Comparison of the PerkinElmer Neonatal Biotinidase kit with the predicate device.
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| Parameter | Neonatal Biotinidase kit | Predicate Device |
|---|---|---|
| Intended Use | The Neonatal Biotinidase kit isintended for the semi-quantitativedetermination of biotinidaseactivity in blood specimens driedon filter paper as an aid inscreening newborns forbiotinidase deficiency. | The Astoria-Pacific®SPOTCHECK Biotinidase 50Hour Reagent Kit (K010844) isintended for the semi-quantitativedetermination of biotinidase, EC3.5.1.12, activity in dried wholeblood spots using the Astoria-Pacific SPOTCHECK® Analyzer.Measurement of biotinidase activityis primarily for the diagnosis andtreatment of biotinidase deficiencyin newborns. This method isintended for in vitro diagnostic useto aid in screening for decreasedlevels of biotinidase activity andnot for monitoring purposes. |
| Specimen type | Whole blood specimen spotted onfilter paper | Same |
| AssayTechnology | Enzymatic | Same |
| Kit content | Calibrators and enzymaticreagents. (additionally includes kitcontrols and microtiter plates) | Same |
| Interpretation ofresults | Calibration curve | Same |
| Test Principle | 1-step enzymatic assay were thebiotinidase in the sample cleavesthe substrate biotin 6-aminoquinoline generating afluorescent 6-aminoquinolineproduct. | 2-step assay were biotinidasereleases p-Aminobenzoic acid(PABA) from biotinyl-p-aminobenzoate. The PABA isdiazotized and coupled to a naptholderivative to form a purplechromophore. |
| Parameter | Neonatal Biotinidase kit | Predicate Device |
| Detectiontechnique | Fluorometric | Colorimetric |
| Instrumentationrequirement | Fluorometer with excitationcentral wavelength of 355 nm andthe emission central wavelengthof 460 nm | Astoria-Pacific SPOTCHECK®Analyzer |
| ScreeningOutcome | Normal and Deficient | Normal, Partial Deficient andProfound Deficient |
| Measuring Unit | U | ERU |
| CalibratorMatrix | Dried Blood Spots prepared fromporcine blood. | Liquid standards. PABA stockstandard. 1.0 mM p-Aminobenzoicacid diluted with Tris Buffer. |
| CalibratorLevels | Six levels, ready to use | Six levels to be prepared fromPABA stock standard |
| 10 U30 U130 U180 U250 U350 U | 0 ERU5 ERU25 ERU50 ERU100 ERU200 ERU | |
| Kit controls | Included in the kit.Dried blood spots prepared fromhuman blood.Normal 275 UAbnormal 50 U | Provided separately |
| Analyticalsensitivity/Lowerlimits ofdetection | Limit of Blank = 12 ULimit of Detection = 16 U | Sensitivity = 1 ERU |
| Linearity | 16 to 390 U | Not defined in predicate labeling |
| MeasuringRange | 16 to 350 U | Not defined in predicate labeling |
| Expected values | Normal PopulationRange: 31.5 - 388 UMean: 163.8 UMedian: 160.9 U | Normal PopulationRange: 19 to 121 ERUAverage: 54 ERU |
| Parameter | Neonatal Biotinidase kit | Predicate Device |
| Interference | Neonate albumin levels above normal (2.8 to 4.4 g/dL) can interfere with this test by increasing biotinidase activity. This could result in the misclassification of a patient with a biotinidase result near the cut-off value as 'normal' when in fact, the patient should be classified as 'deficient'. A patient with known or clinically suspected elevated blood albumin concentration should be screened with an alternative method and confirmed according to local requirements for follow-up testing.Kanamycin sulphate, glutathione, sulfamethoxazole, sulfisoxazole, and trimethoprim can interfere with this test by increasing biotinidase activity. This could result in the misclassification of a patient with a biotinidase result near the cut-off value as 'normal' when in fact, the patient should be classified as 'deficient'. Patients or mothers known to have received kanamycin sulphate, glutathione, sulfamethoxazole, sulfisoxazole or trimethoprim should be screened with an alternative method and confirmed according to local requirements for follow-up testing.Gammaglobulin (1.8 g/dL at biotinidase activity level of 55 U and 1.5 g/dL at biotinidase activity levels of 80 U and 160 U) caused a decrease in biotinidase activity. This could result in a false positive result (biotinidase deficient) in normal patients with biotinidase | Sulfonamides react with color developing reagents.Phenytoin, ampicillin, gentamycin sulfate, vitamin K, penicillin G potassium, kanamycin sulphate, adrenocorticotropic hormone, valproic acid and sodium phenobarbital do not interfere at therapeutic concentrations.Samples spiked with up to 2.5 g/dL of combined albumin and globulin do not interfere as protein added above that level increased the response.Samples spiked with up to 100 mg/dL hemoglobin showed no interference.Samples spiked with up to 250 mg/dL of lipids showed no interference. Lipids added above that level decreased the response. |
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| values near the cut-off value.However, no interference was observed with gammaglobulin (6 g/dL) at a biotinidase activity level of 20 U. |
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| Triglycerides (Intralipid at 150 mg/dL for biotinidase activity levels of 55 U and 80 U, 300 mg/dL at biotinidase activity of 20 U and 400 mg/dL at biotinidase activity of 160 U) caused a decrease in biotinidase activity. This could result in a false positive result (biotinidase deficient) in normal patients with biotinidase values near the cut-off value. |
| Biotin (500 ng/dL) at biotinidase activity level of 55 U caused a decrease in biotinidase activity. This could result in a false positive result (biotinidase deficient) in normal patients with biotinidase values near the cut-off value. |
| Ampicillin (0.56 mg/dL), penicillin G potassium (18.75 mg/dL), sodium phenobarbital (5.5 mg/dL), and phenytoin (1.88 mg/dL) at biotinidase activity level of 160 U caused an increase in response, whereas no interference was observed at biotinidase levels of 20 U, 55 U and 80 U. |
| Valproic acid (25 mg/dL) at biotinidase activity levels of 80 U and 160 U caused an increase in response, whereas no interference was observed at biotinidase activity levels of 20 U and 55 U. |
| The following substances were found not to interfere at the concentrations indicated; adreno- |
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| corticotropic hormone (15 ng/dL),ampicillin (2.8 mg/dL forbiotinidase activities of 20 U, 55 Uand 80 U), ascorbic acid (3mg/dL), biotin (500 ng/dL forbiotinidase activities of 20 U, 80 Uand 160 U), conjugated bilirubin(30 mg/dL), unconjugatedbilirubin (20 mg/dL), EDTA (1g/dL), gentamicin sulphate (0.5mg/dL), hemoglobin (200 mg/dL),heparin(37.5 mg/dL), penicillin Gpotassium (25 mg/dL atbiotinidase activities of 20 U, 55 Uand 80 U), phenytoin (2.5 mg/dLfor biotinidase activities of 20 U,55 U and 80 U), sodiumphenobarbital (5.5 mg/dL forbiotinidase activities of 20 U, 55 Uand 80 U) and vitamin K1 (0.2mg/dL). | PrecisionPrecisionWithin-Run, Within-Lot and Total Precision Samplemean (U) 23 54 75 95 144 287 n 108 108 108 108 108 108 Min (U)measured 17 44 50 71 112 217 Max (U)measured 29 72 97 121 175 358 Withinrun SD 1.4 4.4 5.9 7.9 10 21 Withinrun CV% 6.3 8.2 7.9 8.3 7.2 7.4 Within lotSD 2 5.2 7.8 10 14 30 Within lotCV% 8.8 9.7 10 11 9.8 11 Total SD 2.3 5.5 8.4 11 16 35 TotalCV% 9.9 10 11 11 11 12 | PrecisionWithin-Run and Total Precision Deficient(Profound) PartialActivity Normal n 32 44 44 Average BTD*activity (ERU) 0.54 14.6 3.2 Within-Run Precision, SWR SD 0.09 0.47 3.8 CV% 17 3.2 4.7 Total Precision, ST Average 0.54 14.6 79.6 SD 0.30 0.94 4.6 CV%*Biotinidase 56 6.4 5.8 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Method Comparison
The method comparison was determined in accordance with NCCLS document EP9-A2.
The PerkinElmer Neonatal Biotinidase kit was compared with the Astoria-Pacific Biotinidase 50-Hour Reagent kit (K010844). The samples used in the study included 1516 newborn dried blood spot specimen representing US population analyzed as singlicates with the Neonatal Biotinidase kit and with a commercially available assay. The samples consisted of 1496 routine screening specimens and 20 retrospective specimens diagnosed positive for biotinidase deficiency. The cutoffs were determined according to each method's respective labeling (30% of mean ±2 SD for the Neonatal Biotinidase and 37% of the mean for predicate). All of the 20 clinically confirmed deficient samples were positive by both methods. The observed activities are shown in the following figure.
Image /page/6/Figure/3 description: This image is a scatter plot comparing PerkinElmer Biotinidase Activity (U) on the y-axis and Commercially available assay (ERU*) on the x-axis. The plot shows a cluster of data points, with most points concentrated in the lower-left to middle region of the graph. There are also some data points in the upper-right region, indicating a positive correlation between the two assays. The plot also indicates positive samples (n=20).
The evaluation data (n = 1516) with routine and known positive samples. The routine samples are illustrated with (o) and the known positive samples with (v). The cut-offs are presented with solid lines.
- Enzyme response unit (ERU)
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The screening summaries according to each method's respective labeling are presented in the tables below. The screening positives (+) are samples < cut-off and the screening negatives (-) are samples ≥ cut-off.
| Screening resultCommerciallyavailable kit | Screening resultPerkinElmer kit | Totalsubjects | Diagnosedbiotinidasedeficiency | No diagnosedbiotinidasedeficiency |
|---|---|---|---|---|
| + | + | 20 | 20 | 0 |
| + | - | 2 | 0 | 2 |
| - | + | 1 | 0 | 1 |
| - | - | 1493 | 0 | 1493 |
| Total | 1516 | 20 | 1496 |
| Commercially available kit | Total | ||
|---|---|---|---|
| PerkinElmer kit | Positive(< 15.7 ERU) | Negative(≥ 15.7 ERU) | |
| Positive (<58.5U) | 20 | 1 | 21 |
| Negative (≥58.5U) | 2 | 1493 | 1495 |
| Total | 22 | 1494 | 1516 |
The positive percent agreement was 90.9% (20/22) and the overall percent agreement was 99.8% ((20+1493)/1516).
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Image /page/8/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract image of an eagle or bird-like figure with three curved lines representing the wings or body.
Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center - WO66-0609 Silver Spring, MD 20993-0002
Wallac Ov Division of PerkinElmer, Inc. c/o Kay A. Taylor Senior Manager, Regulatory Affairs 8275 Carloway Road Indianapolis, IN 46236
HAR 0 5 2010
Re: K090123
Trade/Device Name: PerkinElmer Neonatal Biotinidase Kit Regulation Number: 21 CFR §862.1118 Regulation Name: Biotinidase Test System Regulatory Class: Class II Product Code: NAK, JIT, JJX Dated: February 9, 2010 Received: February 16, 2010
Dear Ms. Taylor:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration,
.. ...
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).
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If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at (301) 796-5760. For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or ( 301 ) 796-5680 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Signature
Courtney C. Harper, Ph.D. Director Division of Chemistry and Toxicology. Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
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Indication for Use
510(k) Number (if known):
Device Name:
Indication For Use:
PerkinElmer Neonatal Biotinidase Kit
The Neonatal Biotinidase kit is intended for the semiquantitative determination of biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency.
Prescription Use X (21 CFR Part 801 Subpart D) And/Or
Over the Counter Use (21 CFR Part 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)
Carol Benson
Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) K090123
Page 1 of 1
§ 862.1118 Biotinidase test system.
(a)
Identification. The biotinidase test system is an in vitro diagnostic device intended to measure the activity of the enzyme biotinidase in blood. Measurements of biotinidase are used in the treatment and diagnosis of biotinidase deficiency, an inborn error of metabolism in infants, characterized by the inability to utilize dietary protein bound vitamin or to recycle endogenous biotin. The deficiency may result in irreversible neurological impairment.(b)
Classification. Class II (special controls). The special control is sale, distribution, and use in accordance with the prescription device requirements in § 801.109 of this chapter.