(142 days)
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 antitypsin (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. 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.
Alpha 1 antitrypsin (A1AT) Genotyping Test utilizes Luminex xMAP technology. Genomic DNA is extracted from DBS or from human K2-EDTA anticoaqulated whole blood. 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 above mentioned processing steps, and a CD containing the A1AT Genotyping Test ANALYSIS SOFTWARE and other necessary files. Two kit configurations are available: for 48 or for 192 tests (different amounts of the same reagent components are provided in each case).
Here's a breakdown of the acceptance criteria and study information for the A1AT Genotyping Test, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The FDA summary does not explicitly list "acceptance criteria" but rather presents the results of various performance studies. The reported performance implies the acceptance criteria were met for each category.
| Performance Category | Implied Acceptance Criteria (100% is typical for diagnostic concordance studies) | Reported Device Performance |
|---|---|---|
| Analytical Data | ||
| Lot-to-Lot Repeatability | High concordance (e.g., >95%) | 99.7% overall repeatability for Sample Results |
| External Reproducibility | 100% correct results across sites | 100% of correct Sample Results obtained |
| Real-time Stability | Correct Sample Results at specified time points | All samples provided correct Sample Results at every time point (demonstrated 15 months stability) |
| Open Vial Stability | Correct Sample Results at specified time points after opening | All Sample Results obtained at every time point were correct (proved up to 9 months stability after vials opened) |
| Lower Limit of Detection (LoD) | Detection of 95% of replicates | Highest LoD among two lots was 0.0310 ng/µL of DNA |
| DNA Extraction Validation | Correct Sample Results with different extraction methods | All Sample Results obtained were correct with every extraction method |
| Cross-contamination | No detectable cross-contamination leading to incorrect results | No cross-contamination that could result in an incorrect Sample Results was detected |
| Interfering Substances | Correct Sample Results in the presence of specified interfering substances | Correct Sample Results were obtained in every case |
| Comparison Data | ||
| Method Comparison (vs. Sanger Sequencing) | 100% concordance | 100 % concordance between A1AT Genotyping Test and bidirectional Sanger sequencing results |
2. Sample Size Used for the Test Set and Data Provenance
The "test set" for the primary method comparison study consisted of 116 DNA samples.
- Data Provenance:
- Clinical Samples: 66 samples, type not specified (retrospective/prospective, country of origin not specified).
- Genomic DNAs from Cell Lines: 46 samples, type not specified.
- Synthetic DNA Samples: 4 samples, type not specified.
For the External Reproducibility study, 17 samples were used (5 collected in DBS, 11 archived genomic DNA samples, and 1 synthetic sample). These were tested at external sites in the USA (LifeShare Blood Center and Progenika Inc.) and Italy (University of Pavia). This suggests a mix of prospective and retrospective data, depending on the nature of the archived samples.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The ground truth for the test set in the Method Comparison study was established by bi-directional Sanger sequencing. This is a laboratory-based method of genetic sequencing and does not involve human experts in the interpretation of the results to establish ground truth in the same way clinical image or pathology interpretation would. Therefore, the concept of "number of experts" or their "qualifications" is not directly applicable in this context.
4. Adjudication Method for the Test Set
Not applicable. The ground truth was established by bi-directional Sanger sequencing, which is an objective laboratory method, not subject to adjudication by multiple human readers.
5. 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, an MRMC comparative effectiveness study was not done. This device is a diagnostic test for genetic variants, not an AI-assisted interpretation tool for human readers. Its performance is evaluated fundamentally as a standalone test against an established gold standard (Sanger sequencing).
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) Was Done
Yes, a standalone performance study was done. The entire performance data presented (analytical and method comparison) reflects the performance of the A1AT Genotyping Test system (including the assay and its associated software) as a standalone diagnostic tool. The A1AT Genotyping Test ANALYSIS SOFTWARE algorithm converts the allelic variant genotypes into associated alleles.
7. The Type of Ground Truth Used
The primary ground truth used for the method comparison study was bi-directional Sanger sequencing, a molecular biology technique considered a gold standard for genetic sequence verification.
8. The Sample Size for the Training Set
The document does not specify a training set size. As a diagnostic test that relies on PCR and hybridization, it is unlikely to have a "training set" in the machine learning sense. The device's underlying chemistry and software algorithm are likely developed and validated using a different process than data-driven AI models.
9. How the Ground Truth for the Training Set Was Established
Not applicable, as a "training set" in the context of machine learning for an AI algorithm is not explicitly mentioned or implied for this device. The development of the assay and its software would involve standard molecular biology and software engineering validation processes.
§ 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).