The Progenika A1AT genotyping kit is a qualitative, polymerase chain reaction (PCR) and hybridization-based in vitro diagnostic test to be used with the Luminent (with xPONENT software) for the simultaneous detection and identification of 14 allelic variants and their associated alleles found in the Alpha-1 antitrypsin (AIAT) codifying gene SERPINA1. The test is intended for use with genomic DNA extracted from human whole blood samples collected as dry blood spots (DBS) or in K2-EDTA or from human saliva samples collected as buccal swabs using ORAcollect Dx OCD-100. The A1AT allelic variant genotypes and associated allele results, when used in conjunction with clinical findings and other laboratory tests, are intended as an aid in the diagnosis of individuals with A1AT deficiency (A1ATD). The kit is indicated for prescription use only.

Device Description

Alpha 1 antitrypsin (A1AT) Genotyping Test utilizes Luminex xMAP technology. Genomic DNA is extracted from DBS, from human EDTA anticoagulated whole blood or from human saliva samples collected as buccal swabs using ORAcollect-Dx OCD-100. Extracted DNA is amplified and biotinylated by multiplex PCR and PCR products are denatured and hybridized to oligonucleotide probes coupled to color-coded beads. Hybridized DNA is labeled with a fluorescent conjugate and the resulting signal is detected with a Luminex® 200 system. Raw data obtained is processed with the A1AT Genotyping Test ANALYSIS SOFTWARE in order to obtain the final report. The A1AT Genotyping Test ANALYSIS SOFTWARE algorithm converts the allelic variant genotypes into associated alleles, based on the current literature.

The A1AT Genotyping Test Kit is composed of 4 reagent components (A1AT PCR Master Mix, A1AT Beads Master Mix, SAPE, SAPE Dilution Buffer) required to perform all the abovementioned processing steps. The A1AT Genotyping Test ANALYSIS SOFTWARE, instructions for use and other necessary files are uploaded on a Grifols website. Two kit configurations are available: for 48 or 192 tests (different amounts of the same reagent components are provided in each case).

AI/ML Overview

The provided document describes the A1AT Genotyping Test. This 510(k) submission (K211115) is for a modified version of a previously cleared device (K192858), primarily involving a software update. Therefore, much of the performance data refers back to the original submissions (K192858 and K171868).

Here's a breakdown of the acceptance criteria and study information based on the provided text:

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Acceptance Criteria (Predicate or Implied)	Reported Device Performance (Modified A1AT Genotyping Test)
Lower Limit of Detection (LoD)	Predicate: 0.0310 ng/µl DNA. The modified device's LoD of 0.0215 ng/µl is an improvement, suggesting the acceptance criteria is at least equal to or better than the predicate.	0.0215 ng/µl DNA (highest LoD among two lots)
Precision (Lot-to-Lot Repeatability)	Predicate: Overall correct call rate of 99.7% (one M/S sample provided an incorrect result). For the modified device, the acceptance criteria would be 100% correct calls or similar to the predicate.	100% correct calls
Precision (External Reproducibility)	Predicate: 100% correct calls. The modified device refers to K171868 for this, implying the acceptance criteria is 100% correct calls.	Same as predicate (100% correct calls)
Accuracy	Predicate: Accuracy demonstrated with 147 samples using Bi-directional Sanger sequencing as comparator. The modified device refers to K171868 for Method Comparison (whole blood) and K192858 (saliva), suggesting similar high accuracy against a gold standard. The specific acceptance criteria (e.g., % agreement) are not detailed in this section but are implied to be met.	Same as predicate (demonstrated in K171868 and K192858)

2. Sample Sizes and Data Provenance

Test Set Sample Size:
- LoD: 20 replicates of nine DNA dilutions for a "Sample Panel" (total of 180 tests per lot, across two lots used).
- Precision (Lot-to-Lot): Five DNA samples ("Sample Panel") tested in triplicate, with three different reagent lots, by two operators, on six non-consecutive days, alternating between two Luminex instruments (5 samples * 3 replicates * 3 lots * 2 operators * 6 days = 540 tests, plus additional factors for instruments).
- Accuracy: For the predicate device, 147 samples were used. The document refers to K171868 and K192858 for specific method comparison data for the modified device, so the exact number for the modified device's accuracy testing isn't explicitly stated but would be similar to or larger than the predicate's 147.
Data Provenance: Not explicitly stated as "country of origin for data" or "retrospective/prospective." However, the applicant is Progenika Biopharma S.A. based in Spain, suggesting the studies likely occurred in Spain or affiliated regions. The nature of the studies (analytical performance) implies laboratory-based testing, which can be seen as a form of controlled prospective data collection for the device's technical performance.

3. Number of Experts and Qualifications for Ground Truth (Test Set)

The document does not mention the use of experts to establish ground truth for the test set for the analytical performance studies described (LoD, Precision). These studies focus on the device's ability to consistently and accurately detect specific genetic variants based on known DNA samples.
For Accuracy (Method Comparison), the ground truth was established by Bi-directional Sanger sequencing. This is considered a gold standard in genetic sequencing, not typically requiring "experts" to interpret the sequence output, but rather highly skilled laboratory personnel and bioinformaticians.

4. Adjudication Method (Test Set)

Not applicable as the ground truth for analytical performance studies is typically objective (e.g., known DNA concentrations, Sanger sequencing results). The results are compared directly to these objective standards.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) genetic test, which generates objective results (detection of allelic variants). The performance evaluation focuses on the analytical accuracy and precision of the device itself, rather than human interpretation of complex images or clinical scenarios that would necessitate an MRMC study.

6. Standalone Performance Study (Algorithm Only)

Yes, a standalone performance study was done. The entire "Performance Data" section (H) describes the analytical studies to determine the device's capabilities (LoD, precision, stability, accuracy). These are evaluations of the algorithm and the assay itself, demonstrating its standalone performance from DNA input to reported genotype. The device includes "A1AT Genotyping Test ANALYSIS SOFTWARE" which processes raw data to obtain the final report, indicating its role as a standalone algorithm in the diagnostic process once the PCR and hybridization steps are completed.

7. Type of Ground Truth Used

For LoD and Precision: Known DNA samples with established genotypes/concentrations were used.
For Accuracy (Method Comparison): Bi-directional Sanger sequencing was used as the comparator (a gold standard laboratory method) to establish the ground truth for the samples tested.

8. Sample Size for the Training Set

The document does not explicitly state the sample size used for the training set. This is a 510(k) submission for a device with a software update (v1.0.8.16 from v1.0.6.1). The software's algorithm for converting allelic variant genotypes into associated alleles is based on current literature. It's possible that the "training" for such an algorithm is not based on a 'training set' in the machine learning sense, but rather on established scientific knowledge and rules coded into the software. If any machine learning components were present, their training data is not disclosed here.

9. How the Ground Truth for the Training Set Was Established

Given that the document refers to the algorithm converting genotypes into alleles "based on the current literature," the ground truth for the "training" (or more accurately, the ruleset development) of the algorithm would be derived from published scientific literature and established genetic associations between allelic variants and their corresponding alleles for the SERPINA1 gene. There is no indication of a specific "training set" of patient samples with prospectively established ground truth for algorithm development in this context.

Summary

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May 13, 2021

Progenika Biopharma S.A., a Grifols company Diego Tejedor Technical Director Ibaizabal bidea, Edificio 504, Parque Tecnológico de Bizkaia Derio, Bizkaia 48160 Spain

Re: K211115

Trade/Device Name: A1AT Genotyping Test Regulation Number: 21 CFR 866.5130 Regulation Name: Alpha-1-antitrypsin immunological test system Regulatory Class: Class II Product Code: PZH Dated: April 12, 2021 Received: April 14, 2021

Dear Diego Tejedor:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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 note: 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 either 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

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requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Ying (Katelin) Mao, Ph.D. Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K211115

Device Name A1AT Genotyping Test

Indications for Use (Describe)

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)

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Progenika Biopharma, S.A.

Ibaizabal bidea, Edificio 504
Parque Tecnológico de Bizkaia
48160 Derio - Bizkaia - SPAIN
Phone: +34 94 406 45 25
Fax: +34 94 406 45 26
CIF: ESA95091799
www.progenika.com - www.grifols.com

Special 510(k) Summary

A. Name of the device: A1AT Genotyping Test
B. Common name: Test for SERPINA1 gene genotyping
C. Regulatory information:
a. Classification: Class II
b. Regulation Section: 21 CFR 866.5130, Alpha-1-antitrypsin immunological test system
c. Classification Product Code: PZH, SERPINA1 Variant Detection System
d. Review panel: Immunology (82)
D. Applicant: Progenika Biopharma S.A.

Address: Parque Tecnológico de Bizkaia Ibaizabal bidea, Edificio 504 C.P. 48160, Derio - Bizkaia (Spain) Telephone number: +34 94 406 45 25 Fax number: +34 94 406 45 26 Contact person: Diego Tejedor, Technical Director e-mail: diego.tejedor@grifols.com

E. Intended Use:

The Progenika A1AT genotyping kit is a qualitative, polymerase chain reaction (PCR) and hybridization-based in vitro diagnostic test to be used with the Luminex 200TM instrument (with xPONENT® software) for the simultaneous detection and identification of 14 allelic variants and their associated alleles found in the Alpha-1 antitrypsin (A1AT) codifying gene SERPINA1. The test is intended for use with genomic DNA extracted from human whole blood samples collected as dry blood spots (DBS) or in K2-EDTA or from human saliva samples collected as buccal swabs using ORAcollect·Dx OCD-100. The A1AT allelic variant

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www.progenika.com - www.grifols.com

genotypes and associated allele results, when used in conjunction with clinical findings and other laboratory tests, are intended as an aid in the diagnosis of individuals with A1AT deficiency (A1ATD).

The kit is indicated for prescription use only.

F. Device Description:

G. Substantial Equivalence Information:

Predicate Device: A1AT Genotyping Test 510(k) number: K192858 Applicant: Progenika Biopharma S.A.

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Progenika Biopharma, S.A. lbaizabal bidea. Edificio 504 Parque Tecnológico de Bizkaia 48160 Derio - Bizkaia - SPAIN Phone: +34 94 406 45 25 Fax: +34 94 406 45 26 CIF: ESA95091799 www.progenika.com - www.grifols.com

Main conclusion: The similarities among the candidate device and the predicate device show that A1AT Genotyping Test to be used together with the updated software application is substantially equivalent to the predicate.

Based on the risk assessment and performance data, it can be considered that the differences due to the software update do not raise different questions of safety and effectiveness.

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Progenika Biopharma, S.A.

lbaizabal bidea, Edificio 504 Parque Tecnológico de Bizkaia
48160 Derio - Bizkaia - SPAIN
Phone: +34 94 406 45 25
Fax: +34 94 406 45 25
Fax: +34 94 406 45 26 ClF: ESA95091799
www.progenika.com - www.grifols.com

	Candidate Device	Predicate Device
Item	Modified A1AT Genotyping Test	A1AT Genotyping Test (K192858)
Intended Use	The Progenika A1AT genotyping kit is a qualitative, polymerasechain reaction (PCR) and hybridization-based in vitro diagnostic testto be used with the Luminex 200™ instrument (with xPONENT®software) for the simultaneous detection and identification of 14allelic variants and their associated alleles found in the Alpha-1antitrypsin (A1AT) codifying gene SERPINA1 . The test is intendedfor use with genomic DNA extracted from human whole bloodsamples collected as dry blood spots (DBS) or in K2-EDTA or fromhuman saliva samples collected as buccal swabs usingORAcollect-Dx OCD-100. The A1AT allelic variant genotypes andassociated allele results, when used in conjunction with clinicalfindings and other laboratory tests, are intended as an aid in thediagnosis of individuals with A1AT deficiency (A1ATD).The kit is indicated for prescription use only	The Progenika A1AT genotyping kit is a qualitative, polymerase chainreaction (PCR) and hybridization-based in vitro diagnostic test to beused with the Luminex 200™ instrument (with xPONENT® software)for the simultaneous detection and identification of 14 allelic variantsand their associated alleles found in the Alpha-1 antitrypsin (A1AT)codifying gene SERPINA1 . The test is intended for use with genomicDNA extracted from human whole blood samples collected as dryblood spots (DBS) or in K2-EDTA or from human saliva samplescollected as buccal swabs using ORAcollect-Dx model OCD-100. TheA1AT allelic variant genotypes and associated allele results, whenused in conjunction with clinical findings and other laboratory tests, areintended as an aid in the diagnosis of individuals with A1AT deficiency(A1ATD).The kit is indicated for prescription use only
Specimen Type	Same	Genomic DNA extracted from human whole blood samples collectedas DBS or in K2-EDTA and human saliva samples collected as buccalswabs using ORAcollect-Dx model OCD-100.
Target	Same	Qualitative identification of A1AT alleles (which represent thephenotypes) causing A1ATD.
Item	Candidate Device	Predicate Device
	Modified A1AT Genotyping Test	A1AT Genotyping Test (K192858)
Devicecomponents	Same components, except for the CD component. A1ATGenotyping Test ANALYSIS SOFTWARE and instructions for useare uploaded on a website.	The test is composed of four reagent components (A1AT PCR MasterMix, A1AT Beads Master Mix, SAPE and SAPE Dilution Buffer) insufficient quantity for either 48 or 192 tests and a CD containing theA1AT Genotyping Test ANALYSIS SOFTWARE.
Technology	Same	Polymerase chain reaction (PCR) and hybridization-based in vitrodiagnostic test to be used with the Luminex 200™ instrument (withxPONENT® software).DNA is extracted, amplified and biotinylated by multiplex PCR andPCR products are denatured and hybridized to oligonucleotide probescoupled to color coded beads. Hybridized DNA is labeled with afluorescent conjugate and resulting signal is detected with a Luminex®200 system. The raw data obtained is finally processed with the A1ATGenotyping Test ANALYSIS SOFTWARE in order to obtain the finalreport.
Softwareapplication	The software version has been updated (v1.0.8.16). Results ofevery study have been re-analyzed with this updated softwareversion to ensure acceptance criteria are met.	A1AT Genotyping Test ANALYSIS SOFTWARE v1.0.6.1
Item	Candidate Device	Predicate Device
	Modified A1AT Genotyping Test	A1AT Genotyping Test (K192858)
ProductStability	Same	Real time stability: 24 months at 2-8°COpen vial stability: 9 months at 2-8°C
Performancespecifications	- Lower Limit of Detection: 0.0215 ng/µl DNA.	- Lower Limit of Detection: 0.0310 ng/µl DNA.
	- Interferences: Same	Interferences:
		• For blood samples: hemoglobin, bilirubin, triglycerides andshort blood draw.
		• For saliva samples: α-amylase, hemoglobin,immunoglobin A, total protein, microbes, eating foodwithout beef, eating food with beef, drinking, smoking,chewing gum, mouth washing and brushing teeth.
	- Precision:• Lot-to-Lot: 100% correct calls	Precision:
	• External reproducibility: Same	• Lot-to-Lot: overall correct call rate of 99.7% (one M/S sampleprovided an incorrect result).• External reproducibility: 100% correct calls.
	- Accuracy: Same	- Accuracy: 147 samples, comparator: Bi-directional Sangersequencing.
Intendedpopulation	Same	Prescription use only
DNA extractionmethod	Whole blood samples collected in K2-EDTA: same	Whole blood samples collected in K2-EDTA: QIAamp DNA Blood MiniKit (Qiagen)Whole blood samples collected as DBS:
Item	Candidate DeviceModified A1AT Genotyping Test	Predicate DeviceA1AT Genotyping Test (K192858)
	Whole blood samples collected as DBS: same, although the home-made buffer will be removed.	– Commercial lysis and neutralization solutions (Sigma)– Home-made buffer
	Saliva samples: same	Saliva samples:– QIAamp DNA Blood Mini Kit (Qiagen)– Commercial lysis and neutralization solutions (Sigma)– QIAsymphony DNA Mini Kit (Qiagen)
SpecimenStability	Same	– Whole blood samples collected in K2-EDTA: up to 24 days before DNA extraction at 2-8°C.– Whole blood samples collected as DBS: up to 6 months at ambient temperature.– Saliva samples collected as buccal swabs using ORAcollect-Dx model OCD-100 (DNA Genotek): up to 60 days when stored at ambient temperature.

Comparison table:

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Progenika Biopharma GRIFOLS

Progenika Biopharma, S.A.

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Progenika Biopharma GRIFOLS

Progenika Biopharma, S.A.

lbaizabal bidea, Edificio 504
Parque Tecnológico de Elizkaia
48160 Derio Bizkaia de Sizkaia
48160 Derio Bizkaia estis SPAIN
Fax: +34 94 406 45 25 ClF: ESA95091799
www.progenika.com - www.grifols.com

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Progenika Biopharma GRIFOLS

Progenika Biopharma, S.A.

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H. Performance Data:

Analytical Data:

a) Precision/Reproducibility:
Lot-to-lot repeatability: The lot-to-lot repeatability was determined by testing the "Sample Panel" (five DNA samples covering Z/Z, M/Z, S/S, M/S, and S/Z Sample Results) in triplicate with three different reagent lots, by two operators, on six non-consecutive days, alternating between two Luminex instruments. An overall repeatability of 100% was obtained for Sample Results.

External Reproducibility: See K171868.

b) Reagent Stability:
See K171868 for initial Real-Time and Open-Vial Stabilities information and study designs and K192858 for final claimed stabilities of 24 months reagent stability when stored at 2-8ºC and up to 9 months reagent stability after the vials were first opened.
c) Specimen Stability:
See K171868 for whole blood samples collected as DBS or in K2-EDTA stabilities.

See K152464 and K192858 for saliva samples stability collected in ORAcollect-Dx OCD-100.

d) Lower Limit of Detection (LoD):
The DNA concentration at which 95% of sample replicates resulted in correct Sample Results was determined by testing 20 replicates of nine DNA dilutions of the "Sample Panel" (from 0.16 to 0.0033 ng/μl) using two reagent lots. It was shown that the highest LoD among the two lots used was 0.0215 ng/μl.

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e) DNA Extraction Variability:
See K171868 for DNA Extraction Variability in whole blood samples collected as DBS or in K2-EDTA information.

See K192858 for DNA Extraction Variability in saliva samples.

f) Cross-reactivity and Cross-contamination: See K171868 for Cross-reactivity and Cross-contamination information.
g) Interfering Substances:

See K171868 for Interfering Substances information in whole blood samples collected as DBS or in K2-EDTA.

See K192858 for Interfering Substances information in Saliva samples collected in ORAcollect-Dx OCD-100.

See K192858 for information about potentially interfering variants.

Comparison Data:

h) Method Comparison:
See K171868 for Method Comparison information in whole blood samples. See K192858 for Method Comparison information in saliva samples.

Proposed Labeling: l.

The labeling is sufficient and it satisfies the requirements of 21 CFR 809.10.

J. Conclusion:

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

K. Date of summary preparation:

April 29, 2021

Regulation Number and Section

§ 866.5130

Alpha -1-antitrypsin immunological test system.(a)
Identification. Analpha -1-antitrypsin immunological test system is a device that consists of the reagents used to measure by immunochemical techniques thealpha -1-antitrypsin (a plasma protein) in serum, other body fluids, and tissues. The measurements aid in the diagnosis of several conditions including juvenile and adult cirrhosis of the liver. In addition,alpha -1-antitrypsin deficiency has been associated with pulmonary emphysema.(b)
Classification. Class II (performance standards).