(207 days)
HER2/ER/PR IHControls® -Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level H are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor type (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level M are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® - Level H, HER2/ER/PR IHControls® - Level M and HER2/ER/PR IHControls® - Level L (HER2/ER/PR IHControls®) are immunostaining positive controls for diagnostic immunohistochemistry laboratories. The same immunohistochemical reaction that occurs on a tissue or cellular sample also occurs on the HER2/ER/PR IHControls® microbead. As a result of immunostaining, the microbead bearing the analyte turns the same color as cells expressing the analyte.
HER2/ER/PR IHControls® incorporate assay controls for human epidermal growth factor receptor type II (HER-2), estrogen receptor (ER), and progesterone receptor (PR). The HER2/ER/PR panel includes controls at 3 different analyte concentrations, providing users with an opportunity to select the control with an analyte concentration within the dynamic range of their assay.
HER2/ER/PR IHControls® are provided in a small vial containing enough liquid suspension for at least 100 tests. The user is instructed to pipet a one microliter droplet onto the slide that also bears the patient sample. The droplet is a suspension containing microscopic glass microbeads that are approximately the same size as cells. The microbead surface bears the molecule(s) being measured. Peptides representing the epitopes of all of the major commercial HER2. ER. and PR antibodies used by clinical immunohistochemistry laboratories are incorporated into the products.
HER2/ER/PR IHControls® are comprised of two different microbeads: analyte-coated glass test microbeads (7-8 um in diameter) and color standard microbeads (4.5 um in diameter). The latter provide a fixed brown color intensity for use in standardizing stain intensity measurement.
The HER2/ER/PR IHControls® panel is provided at 3 different analyte concentrations. In descending order of concentration, these levels are denoted "H", "M", and "L". These different products allow matching the analyte concentration to the dynamic range of the laboratory's stain.
Level H (high): This product includes analytes for HER2, ER, and PR, at approximately 10^6 molecules per microbead.
Level M (medium): This product includes analytes for HER2, ER, and PR, at approximately 10^5 molecules per microbead.
Level L (low): This product includes analytes for HER2, ER, and PR, at approximately 10^4 molecules per microbead.
These concentrations are approximate; they are not intended as assayed controls.
Three product levels are provided to allow the user to select a control with the lowest analyte concentration that still produces an easily visible immunostain.
The immunoreactivity pattern for product levels H, M, and L is described in Table 1. The table indicates the primary antibodies corresponding with up to 9 separate HER2/ER/PR peptides. (Some peptides are immunoreactive with more than one primary antibody.) Level H has a narrower range of primary antibody immunoreactivity than levels M and L but the concentrations per microbead are higher. Although any individual primary antibody will be listed as potentially immunoreactive with 2 or more levels. IHC laboratories should use the lowest level that provides for an easily visible stain.
Users pipette a droplet onto a microscope slide that also bears a patient's tissue sample. The droplet hardens upon drying, adheres the analyte-coated microbeads to the microscope slide, and is then processed with the patient sample. The hardened droplet is able to pass through dry heat associated with "baking" of slides, organic solvents associated with deparaffinization, boiling associated with antigen retrieval, and repeated rinses associated with immunostaining.
Here's an analysis of the provided text, outlining the acceptance criteria and study details for the HER2/ER/PR IHControls®:
The document describes the Boston Cell Standards HER2/ER/PR IHControls® a class II medical device (Product Code NJW) intended to monitor the performance of immunohistochemical (IHC) staining processes for HER2, ER, and PR in formalin-fixed paraffin-embedded (FFPE) breast tumor samples. These controls are not for scoring stained slides and are meant to be additional controls, not replacements for existing ones.
Acceptance Criteria and Device Performance
The acceptance criteria for this device are implicitly derived from the analytical performance studies demonstrating specificity, shelf life, and reproducibility of pathologist readouts, as well as the clinical performance studies assessing concordance with conventional tissue controls.
| Acceptance Criteria Category | Specific Criteria (Implicit/Explicit) | Reported Device Performance |
|---|---|---|
| Specificity | No cross-reactivity with antigenically irrelevant primary antibodies. Immunoreactivity with appropriate ER, HER2, and PR primary antibodies. | No cross-reactivity detected with 26 antigenically irrelevant, commonly used primary antibodies. Positive reactivity demonstrated with appropriate ER, HER2, and PR primary antibodies. |
| Shelf Life | Maintain performance for a specified duration when stored appropriately (2-8°C). | All three levels (H, M, L) demonstrated a shelf life of 392 days when stored at 2-8°C. |
| Reproducibility of Readouts | Pathologists should be able to visually interpret the controls and reproducibly render "Pass" or "Fail" scores. | Overall agreement among three pathologists for the interpretation of HER2/ER/PR IHControls® was 100% across all 30 blinded slides (10 for each marker: HER2, ER, PR). This indicates highly reproducible "Pass" or "Fail" scoring. |
| Clinical Concordance | Performance of IHControls® should be highly concordant with conventional tissue controls in assessing IHC test quality in clinical laboratory settings. | Overall concordance rate of 99.5% (440 out of 442 tests) across three clinical sites for combined ER, PR, and HER2 tests. Individual site concordances were 100%, 99.5%, and 99.1%. Two discrepancies were attributed to operator error; one site had a technologist error in pipetting. The device provides "an equally accurate assessment of IHC test quality" compared to tissue controls. |
Study Details
1. Sample Sizes and Data Provenance
- Test Set (Analytical Studies):
- Specificity: Tests against 26 antigenically irrelevant primary antibodies + 3 relevant primary antibodies. (Total 29 tests, implied multiple runs for each to confirm results, but specific number of samples/runs per antibody not detailed.)
- Shelf Life: Real-time testing over approximately 2 years for each of the Level H, M, and L products. (Specific number of samples/tests at different time points not detailed.)
- Reproducibility of Pathologist Readouts: 30 stained HER2/ER/PR IHControl® slides (10 each for HER2, ER, and PR).
- Test Set (Clinical Performance Studies):
- Total Slides: 442 tests/slides across three clinical sites.
- Site 1: 39 (ER) + 39 (PR) + 39 (HER2) = 117 slides
- Site 2: 172 (PR) + 42 (HER2) = 214 slides (Note: 182 tested initially for PR, 172 informative)
- Site 3: 37 (ER) + 37 (PR) + 37 (HER2) = 111 slides
- Data Provenance: The studies were conducted at "three clinical immunohistochemical laboratories." The country of origin is not explicitly stated but is implied to be within the US given the FDA submission. The studies appear to be prospective as the device was "incorporated as on-slide controls... but otherwise followed their typical method for patient testing."
- Total Slides: 442 tests/slides across three clinical sites.
2. Number of Experts and Qualifications for Ground Truth
- Reproducibility of Pathologist Readouts:
- Number of Experts: Three pathologists.
- Qualifications: "Pathologists" are mentioned; specific years of experience or board certifications are not provided, but it's implicit they are qualified to interpret IHC staining.
- Clinical Performance Studies:
- Number of Experts: Not explicitly stated, but "the pathologist interpreted both the conventional (tissue) control and the HER2/ER/PR IHControls®" at each of the three clinical sites. This implies at least one pathologist per site, likely more.
- Qualifications: "Pathologist" is mentioned; specific qualifications are not detailed.
3. Adjudication Method for the Test Set
- Reproducibility of Pathologist Readouts: No explicit adjudication method (e.g., 2+1, 3+1). The study reports 100% concordance among the three pathologists for all slides, indicating no disagreement required adjudication.
- Clinical Performance Studies: No explicit adjudication method for reconciling discrepancies between the IHControls® and tissue controls. The document states discrepancies were "due to operator error."
4. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
- Was it done?: No, an MRMC comparative effectiveness study in the typical sense (comparing human readers with AI vs. without AI assistance) was not performed.
- This device is a quality control material, not an AI diagnostic tool. The "readers" here are pathologists interpreting the performance of the IHC staining process using the control material, not interpreting patient cases.
- The study did involve multiple readers (pathologists) on multiple cases (slides with controls) to assess the reproducibility of the control readout, and compared the control's performance to conventional tissue controls.
- Effect Size: Not applicable, as this was not an AI assistance study.
5. Standalone Performance Study
- Was it done?: Yes, there was an assessment of the device's inherent "standalone" performance in terms of its ability to be correctly recognized as "Pass" or "Fail" by pathologists and its concordance with traditional tissue controls. However, this is "standalone" in the context of a quality control device proving its efficacy, not in the typical AI sense of an algorithm-only performance measurement.
- The "Reproducibility of Pathologist IHControl® Readouts" section assesses the direct interpretation of the IHControls® by pathologists without direct comparison to patient samples or other controls during the reading.
- The "Clinical Performance Studies" assess the IHControls® in conjunction with the IHC staining process and compare their outcome to conventional controls.
6. Type of Ground Truth Used
- Specificity: The ground truth for specificity was established by applying the device to antibodies known not to react with HER2, ER, or PR, and confirming the absence of staining. Conversely, ground truth for intended reactivity was established by applying the device to known HER2, ER, and PR antibodies and confirming positive staining. This relies on established knowledge of antibody-antigen reactions and positive tissue controls.
- Reproducibility of Pathologist Readouts: The "Expected Result" (Pass/Fail) for each of the 30 slides was presumably established by the study design (e.g., intentionally diluted primary antibody to create "Fail" results). This is a designed ground truth based on experimental conditions.
- Clinical Performance Studies: The ground truth for the "quality control check" of the IHC stain was the conventional tissue control (scored as "Pass" or "Fail"). The study implicitly uses the established performance of tissue controls as the gold standard against which the new IHControls® are compared for concordance.
7. Sample Size for Training Set
This information is not provided in the document. This device is a control material for IHC staining, not a diagnostic AI algorithm that typically has a "training set." The development of the device (e.g., optimizing peptide concentrations) is a manufacturing and R&D process, not a machine learning training process.
8. How Ground Truth for Training Set Was Established
Not applicable, as a concept of a "training set" in the machine learning sense is not relevant for this device description. The "training" for the device's design would involve material science, peptide chemistry, and immunohistochemistry expertise.
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August 15, 2022
Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: on the left, there is a seal with an eagle emblem, and on the right, there is the text "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue. The text is arranged in three lines, with "FDA" in a larger font size and a blue square behind it.
Boston Cell Standards, Inc. % Gail Radcliffe, President Radcliffe Consulting, Inc. 23 Fairbanks St West Boylston, MA 01583
Re: K220163
Trade/Device Name: HER2/ER/PR IHControls® - Level H HER2/ER/PR IHControls® - Level M HER2/ER/PR IHControls® - Level L Regulation Number: 21 CFR 864.1860 Regulation Name: Immunohistochemistry Reagents And Kits Regulatory Class: Class II Product Code: NJW Dated: January 10, 2022 Received: January 20, 2022
Dear Gail Radcliffe:
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,
Shyam Kalavar Deputy Branch Chief Division of Molecular Genetics and Pathology OHT7: Office of In Vitro Diagnostics 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) K220163
Device Name Her2/ER/PR IHControls® - Level H Her2/ER/PR IHControls® - Level M Her2/ER/PR IHControls® - Level L
Indications for Use (Describe)
HER2/ER/PR IHControls® -Level H
HER2/ER/PR IHControls® -Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level H are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level M
HER2/ER/PR IHControls® -Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast turnor samples.
HER2/ER/PR IHControls® -Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level M are an additional controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® -Level L
HER2/ER/PR IHControls® -Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® -Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® -Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
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)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
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5. 510(k) Summary
5.1 Boston Cell Standards (BCS)
Sponsor:
Boston Cell Standards Inc. 800 Washington St. Boston, MA 02111 Tel: 617-636-8042
Contact Person:
Steven Bogen, MD, PhD Boston Cell Standards Inc. 800 Washington St. Boston MA 02111 E-mail: sbogen@bostoncellstandards.com Tel: 617-636-5422 FAX: 617-674-3570
Consultant:
Gail Radcliffe, PhD Radcliffe Consulting, Inc. 231 Fairbanks St. West Boylston, MA 01583 E-mail: gradcliffe@RMDCI.com Tel: 508-934-9673
5.2 Device
Trade Name: HER2/ER/PR IHControls® - Level H HER2/ER/PR IHControls® - Level M HER2/ER/PR IHControls® - Level L Common name: Control Material, peptide-based controls for HER2, ER, and PR, Immunohistochemistry Regulation number: 864.1860 Product Code: NJW Class: 2
5.3 Predicate Device
QCS HER2 Immunocontrols (K023335)
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5.4 Intended Use
HER2/ER/PR IHControls® - Level H
HER2/ER/PR IHControls® - Level H are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level H are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level H are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® - Level M
HER2/ER/PR IHControls® - Level M are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level M are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level Mare an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
HER2/ER/PR IHControls® - Level L
HER2/ER/PR IHControls® - Level L are peptide based qualitative on-slide controls to monitor the performance of the analytic components (antigen retrieval and immunostaining) of the immunohistochemical (IHC) staining process for certain human epidermal growth factor receptor type II (HER2), estrogen receptor (ER) and progesterone receptor (PR) IHC stains. It is indicated for use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.
HER2/ER/PR IHControls® - Level L are not intended to be used for scoring HER2, ER, and PR IHC stained slides.
HER2/ER/PR IHControls® - Level L are an additional control to the run controls specified in the HER2, ER, or PR IHC device labeling and are not intended to replace the controls approved or cleared as part of an IHC device.
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5.5 Device Description
HER2/ER/PR IHControls® - Level H, HER2/ER/PR IHControls® - Level M and HER2/ER/PR IHControls® - Level L (HER2/ER/PR IHControls®) are immunostaining positive controls for diagnostic immunohistochemistry laboratories. The same immunohistochemical reaction that occurs on a tissue or cellular sample also occurs on the HER2/ER/PR IHControls® microbead. As a result of immunostaining, the microbead bearing the analyte turns the same color as cells expressing the analyte.
HER2/ER/PR IHControls® incorporate assay controls for human epidermal growth factor receptor type II (HER-2), estrogen receptor (ER), and progesterone receptor (PR). The HER2/ER/PR panel includes controls at 3 different analyte concentrations, providing users with an opportunity to select the control with an analyte concentration within the dynamic range of their assay.
HER2/ER/PR IHControls® are provided in a small vial containing enough liquid suspension for at least 100 tests. The user is instructed to pipet a one microliter droplet onto the slide that also bears the patient sample. The droplet is a suspension containing microscopic glass microbeads that are approximately the same size as cells. The microbead surface bears the molecule(s) being measured. Peptides representing the epitopes of all of the major commercial HER2. ER. and PR antibodies used by clinical immunohistochemistry laboratories are incorporated into the products.
HER2/ER/PR IHControls® are comprised of two different microbeads: analyte-coated glass test microbeads (7-8 um in diameter) and color standard microbeads (4.5 um in diameter). The latter provide a fixed brown color intensity for use in standardizing stain intensity measurement.
The HER2/ER/PR IHControls® panel is provided at 3 different analyte concentrations. In descending order of concentration, these levels are denoted "H", "M", and "L". These different products allow matching the analyte concentration to the dynamic range of the laboratory's stain.
| Level H (high) | This product includes analytes for HER2, ER, and PR, at approximately$10^6$ molecules per microbead. |
|---|---|
| Level M (medium) | This product includes analytes for HER2, ER, and PR, at approximately$10^5$ molecules per microbead. |
| Level L (low) | This product includes analytes for HER2, ER, and PR, at approximately$10^4$ molecules per microbead. |
These concentrations are approximate; they are not intended as assayed controls.
Three product levels are provided to allow the user to select a control with the lowest analyte concentration that still produces an easily visible immunostain.
The immunoreactivity pattern for product levels H, M, and L is described in Table 1. The table indicates the primary antibodies corresponding with up to 9 separate HER2/ER/PR peptides.
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(Some peptides are immunoreactive with more than one primary antibody.) Level H has a narrower range of primary antibody immunoreactivity than levels M and L but the concentrations per microbead are higher. Although any individual primary antibody will be listed as potentially immunoreactive with 2 or more levels. IHC laboratories should use the lowest level that provides for an easily visible stain.
| Level H | Level M | Level L |
|---|---|---|
| (BRLS11) | (BRL2U04) | (BRL2W08) |
| HER2 CB11 | HER2 CB11 | HER2 CB11 |
| HER2 4B5 | HER2 4B5 | HER2 4B5 |
| HercepTest | HercepTest | HercepTest |
| PR 636 | PR 636 | PR 636 |
| PR 16 | PR 16 | PR 16 |
| ER EP1 | ER EP1 | ER EP1 |
| ER 1D5/2.123 | ER 1D5/2.123 | |
| ER 6F11 | ER 6F11 | |
| PR 1294 | PR 1294 | |
| PR 1E2 | PR 1E2 | |
| ER SP1 | ER SP1 |
| Table 1. HER2/ER/PR IHControls® Immunoreactivity: Levels H. M. and L | ||||||
|---|---|---|---|---|---|---|
| ---------------------------------------------------------------------- | -- | -- | -- | -- | -- | -- |
Users pipette a droplet onto a microscope slide that also bears a patient's tissue sample. The droplet hardens upon drying, adheres the analyte-coated microbeads to the microscope slide, and is then processed with the patient sample. The hardened droplet is able to pass through dry heat associated with "baking" of slides, organic solvents associated with deparaffinization, boiling associated with antigen retrieval, and repeated rinses associated with immunostaining.
5.6 Principle of Operation
The HER2/ER/PR IHControls® contain analytes that are immunoreactive in IHC stains. They are present on every slide. Therefore, negative staining due to analytic error is readily detectable. Like tissue samples, the HER2/ER/PR IHControls® are formalin-fixed during manufacture. Therefore, test microbeads in the HER2/ER/PR IHControls® products may demonstrate improved immunoreactivity after antigen retrieval.
5.7 Substantial Equivalence
The similarities and differences between Boston Cell Standards HER2/ER/PR IHControls® and the predicate (QCS control slides for HER2 IHC) are outlined in the table below.
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Table 2. Comparison of HER2/ER/PR IHControls® Panel to Predicate
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| Controlconfiguration | not intended to be used for scoringHER2, ER, and PR IHC stained slides.Her2/ER/PR IHControls® -Level M arean additional control to the run controlsspecified in the HER2, ER, or PR IHCdevice labeling and are not intended toreplace the controls approved or clearedas part of an IHC device.Her2/ER/PR IHControls® -Level LHer2/ER/PR IHControls® -Level L arepeptide based qualitative on-slidecontrols to monitor the performance ofthe analytic components (antigen retrievaland immunostaining) of theimmunohistochemical (IHC) stainingprocess for certain human epidermalgrowth factor receptor type II (HER2),estrogen receptor (ER) and progesteronereceptor (PR) IHC stains. It is indicatedfor use with formalin-fixed paraffin-embedded (FFPE) breast tumor samples.Her2/ER/PR IHControls® -Level L arenot intended to be used for scoringHER2, ER, and PR IHC stained slides.Her2/ER/PR IHControls® -Level L arean additional control to the run controlsspecified in the HER2, ER, or PR IHCdevice labeling and are not intended toreplace the controls approved or clearedas part of an IHC device. | Breast cancer cell lines (MDA-361, MDA-453, MCF-7) fordetection of HER2. |
|---|---|---|
| Level HThis product includes analytes forHER2, ER, and PR, at approximately106 molecules per microbead. | ||
| Level MThis product includes analytes forHER2, ER, and PR, at approximately105 molecules per microbead. | ||
| Level LThis product includes analytes for |
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| HER2, ER, and PR, at approximately104 molecules per microbead. | ||
|---|---|---|
| Assaycompatibility | Polyclonal and monoclonal IHC stains | Polyclonal and monoclonal IHCstains |
| Analyteconcentration | Three different levels of HER2,ER, PR each at differentconcentration. | Three different cell lines eachexpressing unknownconcentrations of HER2. |
| Differences | ||
| Reagents | Formalin-fixed HER2, ER, PR peptideepitopes covalently attached to glassmicrobeads in a liquid matrix thatadhere to a slide. | Formalin-fixed paraffinembedded breast cancer celllines expressing HER2protein, mounted on slides. |
| Slidecontrols | Controls on every slide, as perregulatory guidelines. | Separate slides for batchcontrol |
5.8 Analytical Performance Studies
5.8.1 Specificity
The HER2/ER/PR IHControls® were tested against twenty-six (26) antigenically irrelevant, commonly used primary antibodies, in the context of clinical IHC testing (Table 3). Positive tissue controls demonstrated that the antigenically irrelevant tests were operating correctly. No cross-reactivity was detected with the HER2/ER/PR IHControls®. When an appropriate ER, HER2 and PR primary antibody was used for staining, HER2/ER/PR IHControls® demonstrated immunoreactivity.
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| Marker | Organ | Antibody | TissueControl | IH Controls® | |
|---|---|---|---|---|---|
| 1. | Bcl-6 | Tonsil | GI191E/A8 | Positive | Negative |
| 2. | C4d | Tonsil | SP91 | Positive | Negative |
| 3. | CD10 | Tonsil | SP67 | Positive | Negative |
| 4. | CD138 | Tonsil | B-A38 | Positive | Negative |
| 5. | CD20 | Tonsil | L26 | Positive | Negative |
| 6. | CD34 | Tonsil | QBEnd/10 | Positive | Negative |
| 7. | CD5 | Tonsil | SP19 | Positive | Negative |
| 8. | CD56 | Tonsil | MRQ-42 | Positive | Negative |
| 9. | CMV | Placenta | CMV | Positive | Negative |
| 10. | Cyclin D1 | Lymphoma | SP4-R | Positive | Negative |
| 11. | Cytokeratin20 | Colon | SP33 | Positive | Negative |
| 12. | Cytokeratin 7 | Lung | SP52 | Positive | Negative |
| 13. | EFGR | Normal Skin | 5B7 | Positive | Negative |
| 14. | Keratin | Skin | 34BE12 | Positive | Negative |
| 15. | Ki-67 | Tonsil | 30-9 | Positive | Negative |
| 16. | Mart-1 | Melanoma | A103 | Positive | Negative |
| 17. | MITF | Melanoma | C5/D5 | Positive | Negative |
| 18. | p53 | p53 | BP53-11 | Positive | Negative |
| 19. | p63 | Prostate | 4A4 | Positive | Negative |
| 20. | Pan Keratin | Appendix | AE1/AE3/PCK26 | Positive | Negative |
| 21. | Podoplanin | Tonsil | D2-40 | Positive | Negative |
| 22. | PSA | Prostate | PSA | Positive | Negative |
| 23. | S100 | S100 | 4C4.9 | Positive | Negative |
| 24. | Somatostatin | Pancreas | Somatostatin | Positive | Negative |
| 25. | TAG 72 | Colon | B72.3 | Positive | Negative |
| 26. | TTF-1 | Thyroid | 8G7G3/1 | Positive | Negative |
| 27. | Breast | Breast ca | ER SP1 | Positive | Positive |
| 28. | Breast | Breast ca | HER2 4B5 | Positive | Positive |
| 29. | Breast | Breast ca | PR 1F2 | Positive | Positive |
Table 3. Specificity Testing - List of Primary Antibodies, Targets, and Test Results
5.8.2 Shelf Life
The shelf life for each HER2/ER/PR IHControl® level, when stored at 2 - 8 ° C, was tested in real time over approximately 2 years. Table 4 summarizes product shelf life.
| Table 4. Product Shelf-Life | |
|---|---|
| Product | Shelf Life (days) |
| Level H (BRLS11) | 392 |
| Level M (BRL2U04) | 392 |
| Level L (BRL2W08) | 392 |
5.8.3 Reproducibility of Pathologist IHControl® Readouts
Three pathologists evaluated a blinded slide set of 30 stained HER2/ER/PR IHControl® slides.
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Ten (10) slides each were stained for HER2, ER, and PR. In each set of ten, 4 were stained with a diluted primary antibody, or no primary antibody, to create weak or absent staining. Overall agreement (Table 5) among pathologists for the interpretation of HER2/ER/PR IHControls® was 100%. Pathologists are able to visually interpret HER2/ER/PR IHControls® and reproducibly render Pass or Fail scores.
| Slide number | Pathologist 1 | Pathologist 2 | Pathologist 3 | Expected Result | Concordance |
|---|---|---|---|---|---|
| HER2 | |||||
| 1 | P | P | P | P | 100% |
| 2 | P | P | P | P | 100% |
| 3 | F | F | F | F | 100% |
| 4 | P | P | P | P | 100% |
| 5 | F | F | F | F | 100% |
| 6 | P | P | P | P | 100% |
| 7 | F | F | F | F | 100% |
| 8 | P | P | P | P | 100% |
| 9 | F | F | F | F | 100% |
| 10 | P | P | P | P | 100% |
| ER | |||||
| 11 | P | P | P | P | 100% |
| 12 | P | P | P | P | 100% |
| 13 | F | F | F | F | 100% |
| 14 | P | P | P | P | 100% |
| 15 | F | F | F | F | 100% |
| 16 | P | P | P | P | 100% |
| 17 | F | F | F | F | 100% |
| 18 | P | P | P | P | 100% |
| 19 | F | F | F | F | 100% |
| 20 | P | P | P | P | 100% |
| PR | |||||
| 21 | P | P | P | P | 100% |
| 22 | P | P | P | P | 100% |
| 23 | F | F | F | F | 100% |
| 24 | P | P | P | P | 100% |
| 25 | F | F | F | F | 100% |
| 26 | P | P | P | P | 100% |
| 27 | F | F | F | F | 100% |
| 28 | P | P | P | P | 100% |
| 29 | F | F | F | F | 100% |
| 30 | P | P | P | P | 100% |
Table 5. Pathologist Reader Reproducibility
5.9 Clinical Performance Studies
The HER2/ER/PR IHControls® were incorporated as on-slide controls for HER2, ER, and/or PR immunostaining at three clinical immunohistochemical laboratories. The laboratories incorporated the HER2/ER/PR IHControls® but otherwise followed their typical method for patient testing. After immunostaining was complete, the laboratories performed an assessment. For each stained slide, the pathologist interpreted both the conventional (tissue) control and the HER2/ER/PR IHControls®, for each of the HER2, ER, and PR stains. The controls (tissue
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controls or HER2/ER/PR IHControls®) were scored as either "Pass" or "Fail", depending on whether the control shows that the test passed or failed the quality control check. Percent concordance is reported.
The overall concordance rate is 440 out of 442 tests, or 99.5%, from all 3 clinical trial sites, for ER, PR, and HER2 tests combined (Table 6). Individual site concordance was 100%, 99.5%, and 99.1%.
Two discrepancies between tissue controls and HER2/ER/PR IHControls® were due to operator error: (1) use of an incorrect HER2/ER/PR IHControl®, and (2) placement of the HER2/ER/PR IHControls® at the slide edge because the patient sample and tissue (comparator) control left no other place. There was also one site where a histotechnologist did not pipette the HER2/ER/PR IHControls® onto the slide.
| Site | Immunostain | Product Tested | # Slides | Concurrence | Deviations |
|---|---|---|---|---|---|
| 1 | ER NCL-L-ER-6F11 | BRL2U04-001 | 39 | 39/39 | 0 |
| 1 | PR NCL-L-PGR-312 | BRLS11-001 | 39 | 39/39 | 0 |
| 1 | HER2 BOND Oracle | BRL2U04-001 | 39 | 39/39 | 0 |
| 2 | PR Agilent M3569 | BRL2U04-001 | 172* | 171/172 | 1 |
| 2 | HER2 PATHWAY | BRLS11-003 | 42 | 42/42 | 0 |
| 3 | ER CONFIRM | BRL2W08-003 | 37 | 36/37 | 1 |
| 3 | PR BOND PA0312 | BRLS11-001 | 37 | 37/37 | 0 |
| 3 | HER2 PATHWAY | BRLS11-001 | 37 | 37/37 | 0 |
Table 6. Data Summary from Five Clinical Sites
- Slides from two days were deemed uninformative and not counts for the difference between 182 slides tested and the denominator in the concurrence column being 172, as there were 5 slides per day that did not generate any data.
The data (Table 6) demonstrate that the HER2/ER/PR IHControls® provide an equally accurate assessment of IHC test quality.
6.0 Conclusions from Nonclinical and Clinical Data
The conclusions drawn from the analytical and clinical data demonstrate that the device is safe and effective for its intended use.
§ 864.1860 Immunohistochemistry reagents and kits.
(a)
Identification. Immunohistochemistry test systems (IHC's) are in vitro diagnostic devices consisting of polyclonal or monoclonal antibodies labeled with directions for use and performance claims, which may be packaged with ancillary reagents in kits. Their intended use is to identify, by immunological techniques, antigens in tissues or cytologic specimens. Similar devices intended for use with flow cytometry devices are not considered IHC's.(b)
Classification of immunohistochemistry devices. (1) Class I (general controls). Except as described in paragraphs (b)(2) and (b)(3) of this section, these devices are exempt from the premarket notification requirements in part 807, subpart E of this chapter. This exemption applies to IHC's that provide the pathologist with adjunctive diagnostic information that may be incorporated into the pathologist's report, but that is not ordinarily reported to the clinician as an independent finding. These IHC's are used after the primary diagnosis of tumor (neoplasm) has been made by conventional histopathology using nonimmunologic histochemical stains, such as hematoxylin and eosin. Examples of class I IHC's are differentiation markers that are used as adjunctive tests to subclassify tumors, such as keratin.(2) Class II (special control, guidance document: “FDA Guidance for Submission of Immunohistochemistry Applications to the FDA,” Center for Devices and Radiologic Health, 1998). These IHC's are intended for the detection and/or measurement of certain target analytes in order to provide prognostic or predictive data that are not directly confirmed by routine histopathologic internal and external control specimens. These IHC's provide the pathologist with information that is ordinarily reported as independent diagnostic information to the ordering clinician, and the claims associated with these data are widely accepted and supported by valid scientific evidence. Examples of class II IHC's are those intended for semiquantitative measurement of an analyte, such as hormone receptors in breast cancer.
(3) Class III (premarket approval). IHC's intended for any use not described in paragraphs (b)(1) or (b)(2) of this section.
(c)
Date of PMA or notice of completion of a PDP is required. As of May 28, 1976, an approval under section 515 of the Federal Food, Drug, and Cosmetic Act is required for any device described in paragraph (b)(3) of this section before this device may be commercially distributed. See § 864.3.