The GSP Neonatal Biotinidase kit is intended for the quantitative in vitro determination of human biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency using the GSP instrument.

Device Description

The GSP Neonatal Biotinidase kit contains sufficient reagents to perform 1152 assays. The GSP Neonatal Biotinidase test system measures biotinidase activity, combining an enzyme reaction with a solid phase time-resolved immunofluorescence assay. The GSP Neonatal Biotinidase assay is based on the ability of the biotinidase enzyme to cleave the amide bond in Eu-labeled biotin. The enzyme reaction is stopped by addition of streptavidin which has high affinity for biotin (either Eu-labeled or free biotin). The streptavidin-biotin complexes are captured by the solid phase monoclonal antibody directed against streptavidin. DELFIA Inducer dissociates the molecules into the solution where the europium fluorescence is measured. The measured fluorescence is inversely proportional to the biotinidase activity of the sample.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided text:

Acceptance Criteria and Device Performance

Acceptance Criteria Category	Specific Criteria	Reported Device Performance
Precision (Variation)	Total variation (CV) for dried blood spot samples across 3 kit lots and 3 GSP instruments.	Total variation ranged from 7.5% to 12.7% CV.
Analytical Sensitivity	Limit of Blank (LoB)	LoB = 9.5 U/dL (95th percentile of blank samples, n=150)
	Limit of Detection (LoD)	LoD = 14.8 U/dL (based on 360 determinations of five low-level samples)
	Limit of Quantitation (LoQ)	LoQ = 14.8 U/dL (lowest activity with total CV ≤ 20%)
Linearity	Demonstrate linearity throughout the measuring range.	Demonstrated linear throughout the measuring range (from 14.8 U/dL to 325 U/dL).
Interference	Ampicillin, sulfisoxazole, glutathione, unconjugated bilirubin, and conjugated bilirubin/triglyceride effects.	Interfering: Ampicillin (≥1.4 mg/dL at low biotinidase, 2.8 mg/dL at high biotinidase), Sulfisoxazole (≥7.5 mg/dL at low biotinidase), Glutathione (>30 mg/dL), Unconjugated bilirubin (10 mg/dL at low biotinidase), Conjugated bilirubin (≥2.5 mg/dL), Triglyceride (≥250 mg/dL). Non-interfering: Adrenocorticotropic hormone, ascorbic acid, biotin, Gammaglobulin, gentamicin sulphate, hemoglobin, human serum albumin, kanamycin sulphate, penicillin G, phenytoin, phenobarbital, sulfmethoxazole, trimethoprim, valporic acid, vitamin K1 (at specified concentrations).
Clinical Performance	Agreement with predicate device for screening newborns for biotinidase deficiency.	Overall percent agreement: 99.6% (CI 99.2% - 99.8%) Positive percent agreement: 92.0% (CI 74.0% - 99.0%) Negative percent agreement: 99.7% (CI 99.3% - 99.9%)
	Classification of confirmed biotinidase deficient samples.	All 20 retrospective confirmed biotinidase deficiency specimens were classified as screening positive by the predicate. The GSP method initially classified 19/20 as positive, with one initially negative specimen testing positive in 4 repeat tests.

Study Details

Sample size used for the test set and the data provenance:
- Clinical Study Test Set: 2008 specimens (1988 routine newborn screening specimens and 20 retrospective confirmed biotinidase deficiency specimens).
- Provenance: This information is not explicitly stated, but it can be inferred that these are human blood specimens from newborn screening programs. The "retrospective confirmed biotinidase deficiency specimens" suggest historical data, implying a retrospective study design for these specific samples. The routine screening specimens would be prospective in nature from a screening program. No specific country of origin is mentioned.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- The document does not explicitly state the number of experts or their qualifications for establishing the ground truth.
- For the "retrospective confirmed biotinidase deficiency specimens," the ground truth is implied to be established through confirmation methods for biotinidase deficiency, but no details on expert involvement are provided.
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- The document does not describe any specific adjudication method for establishing the ground truth of the clinical test set. The term "confirmed biotinidase deficiency specimens" implies that a definitive diagnosis was reached, but the process is not detailed.
- For the one discrepant clinical case, it was subjected to "multiple (4) repeat tests." This could be considered a form of internal adjudication/re-testing rather than external expert consensus.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No MRMC or human-in-the-loop study was conducted or described. This device is an in vitro diagnostic kit, meaning it is an automated assay, not an AI assisting human readers. The comparison is between the new automated device and a predicate manual method.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, the core of the clinical study involved evaluating the performance of the GSP Neonatal Biotinidase kit (an automated instrument-based assay) as a standalone device in comparison to the existing manual predicate device. The results are reported as direct comparisons between the two methods on the same samples.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- The ground truth for the clinical study was based on the classification of specimens as "biotinidase deficient" or "normal." For the 20 known deficient specimens, the ground truth was "retrospective confirmed biotinidase deficiency." For the routine screening specimens, it is implied that the predicate device's result (manual Neonatal Biotinidase method) served as a comparative reference, which itself would have a ground truth based on established clinical diagnostic criteria for biotinidase deficiency. The cut-offs used for both devices (0.5th percentile for GSP and 30% of mean + 2SD for the predicate) are tied to population distributions expected for the condition.
The sample size for the training set:
- The document describes the GSP Neonatal Biotinidase kit as a diagnostic assay, and its development would typically involve internal validation and optimization data. The provided text, being a 510(k) summary, primarily focuses on the test set performance to demonstrate substantial equivalence.
- There is no explicit mention of a separate "training set" in the context of machine learning or AI models, as this is an in vitro diagnostic kit based on an enzymatic reaction. The calibrators and controls used in the kit are standardized and prepared from human whole blood, aiding in the assay's calibration and ongoing quality control during operation.
How the ground truth for the training set was established:
- Not applicable in the context of a "training set" for a traditional in vitro diagnostic assay.
- For the calibrators, they were "calibrated against in-house primary calibrators (dried blood spots, stored at -80 to -60°C) prepared using adult human blood (endogenous biotinidase activity in serum) and washed red blood cells as blood matrices." This describes how the reference values for the assay's internal calibration curve are established.

Summary

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510k Summary

This 510k Summary information is supplied in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is K131284

Date: November 14, 2013

Submitted by:	Wallac Oy, a subsidiary of PerkinElmer Inc.940 Winter StreetWaltham, MA 02451	NOV 14 2013
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Submission Contact Person:	Jeanette Schier-PugsleyDirector, Regulatory and Clinical AffairsTel: 781-663-6025Fax: 781-663-5969
Trade Name:	GSP Neonatal Biotinidase kit
Common Name:	Biotinidase test system
Regulation:	21 CFR 862.1118
Classification Name:	Biotinidase test system
Classification:	II
Panel:	75 Clinical Chemistry
Product Code:	NAK
Predicate device:	PerkinElmer Neonatal Biotinidase kit [K090123]

Device Description:

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The kit contains the following components:

Calibrators have been prepared from human whole blood using ProClin 300 as preservative. The biotinidase calibrators' activities range from 10 U/dl to 325 U/dL. As no internationally accepted reference material or a reference method for the determination of biotinidase activity is available, the calibrators have been calibrated against in-house primary calibrators (dried blood spots, stored at -80 to -60°C) prepared using adult human blood (endogenous biotinidase activity in serum) and washed red blood cells as blood matrices.

Controls have been prepared from human whole blood using ProClin 300 as preservative. The low control is approximately 40.0 U/dL and the high control approximately 160 U/dL.

All human source materials used in the preparation of kit components were tested and found to be negative for hepatitis B surface antigen, anti-hepatitis C and anti-HIV 1 and 2 derivatives by FDA approved methods.

Biotinidase Substrate Reagent - 3 ready-to-use vials, 2.8 ml each Biotinidase SA Reagent - 3 ready-to-use vials, 2.8 ml each Assay Buffer - 3 bottles, 120 ml each Anti-SA Microtitration Strips - 12 plates

Intended Use:

Comparison Chart:

Comparison of the GSP Neonatal Biotinidase kit (proposed device) with its predicate:

Characteristics	-- Proposed Device – GSPNeonatal Biotinidase kit	Predicate (K090123)
IntendedUse/Indications forUse	The GSP Neonatal Biotinidase kitis intended for the quantitative in vitro determination of humanbiotinidase activity in bloodspecimens dried on filter paper asan aid in screening newborns forbiotinidase deficiency using theGSP® instrument.	This kit is intended for the semi-quantitative determination ofbiotinidase in blood specimen driedon filter paper as an aid in screeningnewborns for biotinidase deficiency.
Specimen Type	Dried blood spot	Same
Assay Technology	Enzymatic assay	Same
Kit Content	Calibrators, enzymatic reagents,kit controls (additionally includesmocrotiter plates and bar codelabels for use with the GSPinstrument)	Calibrators, enzymatic reagents, kitcontrols (additionally includesmocrotiter plates)

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Interpretation of Results	Calibration Curve	Same
Test Principle	Combines an enzyme reaction with a solid phase time-resolved immunofluorescence assay. Biotinidase in the sample cleaves the amide bond in the Eu-labeled biotin substrate. The end product is captured using Streptavidin and Anti-streptavidin coated plate. DELFIA® Inducer dissociates the molecules into the solution where the europium fluorescence is measured	1-step enzymatic assay were the biotinidase in the sample cleaves the substrate biotin 6- aminoquinoline generating a fluorescent 6- aminoquinoline product
Instrument Platform	GSP instrument, automated (K090846)	Fluorometer, manual
Detection Method	Time-resolved fluorescence	Fluorometer with excitation central wavelength of 355 nm and the emission central wavelength of 460 nm
Screening Outcome	Normal and Deficient	Same
Measuring Unit	U/dL	U
Calibrator Matrix	Dried blood spots prepared from human whole blood	Dried blood spots prepared from porcine blood
Calibrator Levels	Six levels, ready to use	Six levels, ready to use
	10 U/dL25 U/dL50 U/dL100 U/dL175 U/dL325 U/dL	10 U30 U130 U180 U250 U350 U
Measuring Range	14.8 U/dL - 325 U/dL	16 U - 350 U
Lower Limits of Measure	LoB = 9.5 U/dlLoD = 14.8 U/dlLoQ = 14.8 U/dl	LoB = 12 ULoD = 16 U

Summary of Non-Clinical Studies:

The variation of the GSP Neonatal Biotinidase assay was determined using dried blood spot samples, 3 kit lots, and 3 GSP instruments. The study was performed in 27 runs over 20 days, each run consisting of 2 plates with 4 replicates per sample. Total number of measurements was 216 per sample. Total variation ranged from 7.5 to 12.7 %CV.

The limit of blank, detection and quantity were determined according to the CLSI guideline EP17-A.The Limit of Blank (LoB) for GSP Neonatal Biotinidase kit is 9.5 U/dL, defined as the 95th percentile of a distribution of blank samples (n=150). The Limit of Detection (LoD) is 14.8

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U/dL based on 360 determinations of five low level samples. The Limit of Quantitation (LoO) is 14.8 U/dL, defined as the lowest activity with a total CV equal or less than 20%.

Linearity was determined in accordance with CLSI document EP6-A. For GSP Neonatal Biotinidase, the method has been demonstrated to be linear throughout the measuring range (from 14.8 U/dL to 325 U/dL). Recovery cannot be determined as biotinidase is not commercially available in pure form.

Ampicillin (1.4 mg/dL and above), sulfisoxazole (7.5 mg/dL and above) at low biotinidase activity levels (35 U/dL) and ampicillin (2.8 mg/dL) at high biotinidase activity levels (150 U/dL) were found to interfere with this test by increasing measured biotinidase activity by 19.9%, 32.1% and 15.6%, respectively. Elevated ampicillin and suffisoxazole levels near the biotinidase cut-off did not exhibit a significant effect (<15%).

Glutathione levels above normal (> 30 mg/dL) can interfere with this test by increasing biotinidase activity by 16.1% or more. 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 glutathione concentration (>30 mg/dL)should be screened with an alternative method and confirmed according to local requirements for follow-up testing.

Unconjugated bilirubin (10 mg/dL) added to whole blood at low biotinidase activity levels (35 U/dL) were found to interfere with this test by increasing measured biotinidase activity by 18.7%. Elevated unconjugated bilirubin level (20 mg/dL) near the biotinidase cut-off did not exhibit a significant effect (<15%).

Conjugated bilirubin (2.5 mg/dL and above) and triglyceride (250 mg/dL and above) added to whole blood were found to interfere with this test by decreasing measured biotinidase activity by 26.0% and 15.7%, respectively. Elevated conjugated bilirubin (2.5 mg/dL and above) and triglyceride levels (250 mg/dL and above)may cause a false positive screening result for a specimen with measured biotinidase activity near the cut-off.

The following substances were found not to interfere at concentration indicated: adrenocorticotropic hormone (15 ng/dL), ascorbic acid (3 mg/dL), biotin (500 ng/dL), Gammaglobulin (3 g/dL), gentamicin sulphate (0.5 mg/dL), hemoglobin (1.6 g/dL), human serum albumin (6 g/dL), kanamycin sulphate (3 mg/dL), penicillin G (25 mg/dL), phenytoin (2,5 mg/dL), phenobarbital (5.5 mg/dL), sulfmethoxazole (20 mg/dL), trimethoprim (2 mg/dL), valporic acid (19 mg/dL), and vitamin K1 (0.1 mg/dL).

Summary of Clinical Studies:

The 3307-001U GSP Neonatal Biotinidase kit was compared to the 3018-0010/3018-001B Neonatal Biotinidase kit in a routine screening laboratory by measuring the biotinidase activity in a total of 2008 specimens. These included routine newborn screening specimens (n = 1988) and retrospective confirmed biotinidase deficiency specimens (n = 20).

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The results for the screening performance data based on the predicate's cut-off of 30% of mean + 2SD manual Neonatal Biotinidase method classified all 20 biotinidase deficient specimen as screening positive. The GSP Neonatal Biotinidase method used cut-off based on 0.50 percentile and classified 19 specimens as screening positive and 1 specimen as screening negative based on the initial test result. In multiple (4) repeat tests this specimen was positive at all cut-offs. Duplicate punches in the same microtiter well (due to static electricity) were concluded to be the potential cause for the initial false negative results are shown below.

GSP result	Predicate result	Total	Positive	Normal
+	+	23	19	4
+	-	6	0	6
-	+	2	1*	1
-	-	1977	0	1977
Total		2008	20	1988

" One retrospective confirmed biotinidase deficiency specimen that initially tested as negative; tested as positive in 4 repeat tests.

The following table illustrates the screening performance of the GSP Neonatal Biotinidase test system in comparison to the predicate device using the 0.5" percentile cutoff based on population distribution and using the 30% of mean + 2SD cut-off for manual Neonatal Biotinidase method. The percent agreement calculations have been performed for the comparison.

		3018-0010/3018-001B 30% of mean + 2SD
		Screen positive≤ 67.6 U	Screen negative> 67.6 U	Total
GSP3307-001U0.5thpercentile	Screen positive≤ 74.6 U/dL	23*	6	29
	Screen negative> 74.6 U/dL	2**	1977	1979
	Total	25	1983	2008

Includes 19 retrospective confirmed biotinidase deficiency specimens.

** One retrospective confirmed biotinidase deficiency specimen that initially tested as negative; tested as positive in 4 repeat tests

Overall percent agreement = 99. 6% (CI 99.2% - 99.8%) Positive percent agreement = 92.0% (CI 74.0% - 99.0%) Negative percent agreement = 99.7% (CI 99.3% - 99.9%)

Substantial Equivalency:

The proposed device and predicate device utilize similar enzymatic pathway and design which is shown to produce equivalent screening performance in a clinical setting. Both devices are intended for the quantitative in vitro determination of human biotinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency.

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Conclusion:

The GSP Neonatal Biotinidase test system demonstrates analytical and screening performance that supports its substantial equivalency with the predicate device, the Neonatal Biotinidase test system (K090123).

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WORG-GB()s Silver Spring, MI) 20993-0002

November 14, 2013

Wallac Oy. a subsidiary of PerkinElmer Inc. Ms. Jeanette Schier-Pugsley Director, Regulatory and Clinical Affairs 940 Winter Street WALTHAM MA 02451

Re: K131284

Trade/Device Name: GSP Neonatal Biotinidase Kit Regulation Number: 21 CFR 862.1118 Regulation Name: Biotinidase test system Regulatory Class: II Product Code: NAK Dated: October 03, 2013 Received: October 04. 2013

Dear Ms. Schier-Pugsley:

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. Please notc: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into cither class II (Special Controls) or class III (PMA). it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations. Title 21. Parts 800 to 898. In addition. FDA may publish further announcements concerning your device in the Federal Register.

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 (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act): 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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/ReportalProblem/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) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Carol C. Benson -S for

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.

510(k) Number (if known)

K131284

Device Name

GSP Neonatal Biotinidase Kit Indications for Use (Describe)

The GSP Neonatal Biotinidase kit is intended for the quantitative in vitro determination of human biolinidase activity in blood specimens dried on filter paper as an aid in screening newborns for biotinidase deficiency using the GSP instrument.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

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

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

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Regulation Number and Section

§ 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.