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BK100068 Summary

510K Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

Submitter:One Lambda Incorporated
21001 Kittridge Street
Canoga Park, CA 91303
Phone: 818-702-0042
Fax: 818-702-6904
Contact:Mr. Don Arii
Vice President of Operations
Phone: 818-702-0042 x 124
Email: darii@onelambda.com
Or
Mrs. Angela Estany
Regulatory Affairs Manager
Phone: 818-702-0042 x 218
Email: aestany@onelambda.com
Date:22 Nov 2010
Device Name:HNA Genotyping Tray
Classification/
Device Code:
Unclassified
Predicate Device:Micro SSP™ HLA Class II DNA Typing Kit, BK960062
Predicate Classification/
Device Code:
Unclassified, MZI

Device Descriptions: Human neutrophil antigens (HNA) are involved in a variety of clinical conditions including immune neutropenias, transfusion related acute lung injury (TRALI), refractoriness to granulocyte transfusions and febrile transfusion reactions. The HNA1 system has three known antigens: HNA-1a (NA1), HNA-1b (NA2) and HNA-1c (SH), located on the neutrophil Fcγ receptor IIIb (CD16b). These antigens are defined by multiple nucleotide polymorphisms FcγRIIIb gene. While the molecular basis of the HNA3 antigen system is not well defined, the HNA-3a (5b) and HNA-3b (5a) antigens are located on a 70-95 kDa glycoprotein, now thought to be part of the of the choline transporter-like protein-2 (CTL2)-gene. Finally, the HNA-4a (Mart) and HNA-5a (Ond) antigens were found to be caused by single nucleotide mutations in the αM chain (CD11b) and αL integrin unit (CD11a) subunits of the leukocyte adhesion molecules (β2 integrins), respectively. Historically, serological methodologies have been used for HNA typing. Serological methods can be limited by antibody and type-specific neutrophil availability. With the advent of PCR technologies, DNA-based tissue typing techniques have become routine in the laboratory. For most DNA-based methodologies, the PCR process is used only as an amplification step to acquire the needed target DNA. The typing process then requires a post-amplification step to discriminate between the different alleles (e.g., RFLP, SSOP, reverse dot blot). By contrast, HNA Genotyping Trays provide all necessary reagents to perform Sequence Specific Primer (SSP) PCR on isolated genomic DNA. However, unlike other PCR-based methods, the SSP methodology employed through One Lambda, Inc discriminates between the different polymorphisms during the PCR process. This shortens the post-amplification processing time to a simple gel electrophoresis detection step and the results are either positive or negative. This abolishes the need for complicated interpretation of results.

The HNA Typing Tray provides sequence-specific primers (SSP) for amplification of alleles of the HNA coding genes by the polymerase chain reaction (PCR). The HNA Typing Tray offers the convenience of optimized, pre-aliquoted primers for immediate PCR and electrophoretic analysis of DNA samples.

Pre-optimized sequence specific primers for amplification of HNA genes are presented (dried) in different wells of a thin-walled tube tray for PCR, ready for the addition of the DNA sample, Taq polymerase, and specially formulated dNTP-buffer mix. A qualitative indicator of specific DNA amplification is obtained by the observation of ethidium bromide fluorescence of amplified DNA (in control and test bands) following quick gel electrophoresis of the PCR products.

Assessment of HNA alleles is performed by analysis of the gel-banding pattern using a reaction-pattern typing grid. Each typing test includes a negative control, which detects the presence of any contaminating PCR products generated in the assay.

Operation Principles: The PCR-SSP methodology is based on the principle that completely matched oligonucleotide primers are more efficiently used in amplifying a target sequence than a mismatched oligonucleotide primer by recombinant Taq polymerase. Primer pairs are designed to have perfect matches only with a single allele or group of alleles. Under strictly controlled PCR conditions, perfectly matched primer pairs result in the amplification of target sequences (i.e., a positive result) while mismatched primer pairs do not result in amplification (i.e., a negative result). After the PCR process, the amplified DNA fragments are separated by agarose gel electrophoresis and visualized by staining with ethidium bromide and exposure to ultraviolet light. Interpretation of PCR-SSP results is based on the presence or absence of a specific amplified DNA fragment. Since amplification during the PCR reaction may be adversely affected by various factors (pipetting errors, poor DNA quality, presence of inhibitors, etc.), an internal control primer pair is included in every PCR reaction. The control primer pair amplifies a conserved region of the Human β-globin gene, which is present in all human DNA samples and is used to verify the integrity of the PCR reaction. In the presence of a positive typing band (specific amplification of an HNA allele), the product of the internal control primer pair may be weak or absent due to the differences in concentration and melting temperatures between the specific primer pairs and the internal control primer pair. The amplified DNA fragments of the specific HNA primer pairs are smaller than the product of the internal control primer pair, but larger than the diffuse, unincorporated primer band. Thus, a positive reaction for a specific HNA allele or allele group is visualized on the gel as an amplified DNA fragment between the internal control product band and the unincorporated primer band.

Indications for Use: HNA GenotypingTray is used for the molecular determination of neutrophil polymorphisms.

Predicate Device Table:

 PredicateSubstantially Equivalent Device

Device

Micro SSP™ HLA Class II DNA Typing KitHNA Genotyping™ Trays

FDA Device Classification

BK#960062
Unclassified under CBER
Device code -MZI

Unassigned #
New Device

Intended Use

Molecular determination of HLA alleles
Molecular determination of neutrophil polymorphisms-HNA 1a,1b, 1c, 3a, 3b, 4a, 4b, 5a, and 5b

Standards Met

Standards set by ASHI. (American Society of Histocompatibility and Immunogenetics) for certification of clinical HLA laboratories or by CLIA for other clinical laboratories
Standards set by AABB for blood banks or by CLIA for other clinical laboratories

Where Used and Target population

Preliminary clinical testing for detection and identification HLA alleles in potential donors and recipients of bone marrow, tissue, or organ transplants.
Preliminary clinical testing for detection and identification of HNA alleles in potential donors of blood or blood components for transfusion.
Assay Method
Allele specific PCR using sequence specific priming (SSP)
Assay Technology
Separation of PCR products by agrose electrophoresis
Reactive Ingredient HLA specific primersHNA specific primers
Specimen Type
DNA
Controls
Internal Control Primer

Detection Method

Qualitative indicator of specific DNA amplification is obtained by the observation of ethidium bromide fluorescence of amplified DNA (in control and test bands).

Positive Reaction

A positive reaction for a specific allele or allele group is visualized on the gel as an amplified DNA fragment between the internal control product band and the unincorporated primer band.
Evaluation of Results

Visual detection of product bands under UV light

Predicate Device Summary:
HNA Genotyping Tray is substantially equivalent to the predicate devices Micro SSP™. Both products are PCR based molecular assays using sequence specific priming for the detection of polymorphisms on cell surface proteins and use the same technology (gel electrophoresis and staining) to determine molecular alleles for preliminary clinical testing. No new safety or effectiveness issues were raised.

Testing:
The performance of the HNA Genotyping Tray was verified and testing demonstrates safety and effectiveness:

Verification of PrimersAgreement
PCR reaction profile of reference samples vs. HNA specific primers100%
  
Specificity/RobustnessAgreement
Replicate testing between technicians100%
Reduced DNA concentration testing100%
  
Detection LimitAgreement
Reduced DNA concentration testing100%
  
Stability - based on accelerated studies and similarity with predicate device-80 °C to -20 °C for 5 months
  
Lot to Lot ConsistencyAgreement
In house100%
  
Clinical Testing Agreement
Performance100%
  
Reproducibility Agreement
In house100%
Clinical100%

Reference samples using gold standard –sequencing, using -------(b)(4)-----------------------------------------------

Overall Conclusion: Based on the assessment of testing performed, One Lambda Incorporated concludes that HNA Genotyping™ Tray is safe and effective. Submitted information is complete and supports that HNA Genotyping™ Tray is substantially equivalent to Micro SSP™, predicate device.