(151 days)
CINtec® Histology is a qualitative immunohistochemistry (IHC) test using mouse monoclonal anti-p16 antibody clone E6H4, and is intended for use in the light microscopic assessment of the p16INK4a protein in formalin-fixed, paraffinembedded (FFPE) cervical punch biopsy tissues using OptiView DAB IHC Detection Kit on a VENTANA BenchMark ULTRA instrument. The test is indicated as an adjunct to examination of hematoxylin and eosin (H&E) stained slide(s), to improve consistency in the diagnosis of cervical intraepithelial neoplasia (CIN). Diagnosis of CIN presence or level should be based on H&E stained slide(s) and other clinical and laboratory test information.
Intended for in vitro diagnostic (IVD) use. Prescription Use Only.
CINtec® Histology is a single dispenser immunohistochemical (IHC) assay system comprised of an anti-p16 primary antibody optimized for use with the BenchMark ULTRA automated slide staining instrument and the OptiView DAB IHC Detection Kit. The antibody is diluted in a Tris-HCl buffer containing carrier protein and 0.1% ProClin 300 as a preservative and provided as a ready-to-use liquid in a FloLock dispenser. CINtec Histology is available in a 50-test size and a 250-test size.
The OptiView DAB IHC Detection Kit (OptiView) is an indirect, biotin-free system for detecting mouse IgG, mouse IgM, and rabbit IgG primary antibodies and is comprised of 6 dispensers packaged together in one box.
Ancillary reagents required to perform the CINtec Histology assay include EZ Prep, Reaction Buffer, ULTRA High Temperature Liquid Coverslip (LCS), ULTRA Cell Conditioning 1 Solution (CC1), Hematoxylin II Counterstain, and Bluing Reagent.
Positive and negative tissue controls that are fixed and processed in the same manner as the test specimens should be used when performing this test. A negative reagent control mouse monoclonal antibody shall be used to evaluate nonspecific staining.
Here's a breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text.
Device Name: CINtec® Histology
Description: A qualitative immunohistochemistry (IHC) test using mouse monoclonal anti-p16 antibody clone E6H4, intended for use in the light microscopic assessment of the p16INK4a protein in formalin-fixed, paraffin-embedded (FFPE) cervical punch biopsy tissues. It's an adjunct to H&E stained slides for improving consistency in diagnosing cervical intraepithelial neoplasia (CIN).
1. Table of Acceptance Criteria and Reported Device Performance
The provided text focuses on the non-clinical performance evaluation of a recombinant CINtec Histology device compared to a hybridoma predicate device, primarily to show their equivalency. The tests are designed to demonstrate the recombinant device's performance against established criteria.
| Test | Acceptance Criteria | Reported Device Performance |
|---|---|---|
| Western Blot | Single band between 15-20 kDa (~16 kDa) must be detected on the Western blot membrane for those lanes loaded with recombinant p16INK4a protein or with lysates from p16INK4a-expressing cell lines and consequently probed with recombinant anti-p16INK4a antibody or hybridoma anti-p16INK4a antibody. | Pass |
| Peptide Inhibition | Decreased staining in the tissues stained with recombinant CINtec Histology reagent containing p16 epitope-specific peptide when compared to the tissues stained with recombinant CINtec Histology reagent containing diluent or non-specific peptide. Significant p16 signal reduction with highest concentration of p16-specific peptide. Tissues with diluent only must show appropriate specific staining. Middle and lowest concentrations of p16-specific peptide should score between 0 and control. Duplicate samples must stain equivalently (within 0.5 point). Background ≤ 0.5 point for ≥ 90% of samples. Negative control MTB slide should not have any specific staining. | Pass |
| Between Lots precision | Stain intensity shall not vary more than 0.5 point from the median score on a 0-4 scale of each sample on ≥ 85% between lots. All samples shall show ≥ 90% positive/negative agreement for CINtec® Histology status. Antibody shall demonstrate background/cross-reactivity ≤ 0.5 points on a 0-4 scale in ≥ 90% of the tissue samples stained. | Pass |
| Immunoreactivity | Background/cross-reactivity ≤ 0.5 points on a 0-4 scale in ≥ 90% of the tissue samples stained. Recommended staining protocol shall preserve tissue morphology as noted by the qualified reader in a minimum of 90% of interpretable samples stained. | Pass |
| Equivalency/Method Comparison | Overall percent agreement (OPA) shall demonstrate a lower bound for the two-sided 95% confidence interval (LBCI) of ≥ 85%. Background should be ≤ 0.5 points (on a 0-4 scale) in 90% or greater of the tissue samples stained. Tissue morphology should be preserved as noted by the qualified reader in a minimum of 90% of interpretable samples stained. | Pass |
| Stability | Stain intensity for all tissue slides stored at 45°C for at least 227 hours or at 37°C for at least 493 hours shall not vary more than 1.0 point in stain intensity as compared to the respective reference tissue slides stained at 0 hour. Background for all tissue slides stored as above shall not exceed 0.5 points as compared to the respective Time 0 reference slides for 24 months' expiration dating. | Pass |
2. Sample sizes for the test set and data provenance
The document describes non-clinical performance evaluation, not a typical "test set" in the machine learning sense with a distinct training/test split for generalizability. The sample sizes are specific to each validation test:
- Peptide Inhibition: Multi-tissue block (MTB) containing various cervical diagnoses.
- Between Lots precision: 26 cervical cases with various diagnoses.
- Immunoreactivity: Tour of Body (TOB), Tour of Tumor (TOT), and 20 additional cases.
- Equivalency/Method Comparison: 249 cervical cases with various diagnoses.
- Stability: Normal cervix, cervical squamous cell carcinoma (SCC), tonsil.
The data provenance (country of origin, retrospective/prospective) is not specified in the provided text. Given it's a device submission, it's likely retrospective use of archived FFPE tissue blocks.
3. Number of experts and qualifications for ground truth
The document does not specify the number of experts or their qualifications used to establish ground truth for the non-clinical performance evaluation. It mentions "qualified reader" for morphology assessment in Immunoreactivity and Equivalency/Method Comparison studies, implying expert assessment, but no details are provided. For the Indications for Use, it states "Diagnosis of CIN presence or level should be based on H&E stained slide(s) and other clinical and laboratory test information," implying that the final diagnosis relies on established pathology expertise in a clinical setting.
4. Adjudication method
The document does not specify any formal adjudication method (e.g., 2+1, 3+1) for the non-clinical performance studies. The "within 0.5 point" criteria for duplicate samples in the Peptide Inhibition test suggests internal consistency checks rather than multi-reader adjudication of a ground truth.
5. Multi-Reader Multi-Case (MRMC) comparative effectiveness study
The document explicitly states: "The substantial equivalence is not based on an assessment of clinical performance data." This indicates that an MRMC comparative effectiveness study, which would typically assess human reader improvement with AI assistance, was not performed or submitted as part of this specific 510(k) (which focuses on equivalency of recombinant vs. hybridoma antibody).
6. Standalone (algorithm only without human-in-the-loop) performance
The device described is an immunohistochemistry (IHC) test (a laboratory assay), not an AI algorithm. Therefore, "standalone (algorithm only without human-in-the-loop) performance" is not applicable in the typical sense of AI device evaluation. The performance metrics focus on the assay's biochemical and staining characteristics, and its interpretation is intended to be by a human pathologist as an "adjunct to examination of hematoxylin and eosin (H&E) stained slide(s)."
7. Type of ground truth used
For the peptide inhibition, immunoreactivity, and equivalency/method comparison studies, the ground truth implied is the known characteristics of the tissue samples (e.g., tissues with varying levels of p16 expression, normal vs. carcinoma, various diagnoses). The "qualified reader" assessment of morphology and staining intensity serves as a measure against accepted pathology interpretations for those samples.
8. Sample size for the training set
This document pertains to the submission of an IHC assay, not a machine learning or AI device. Therefore, the concept of a "training set" for an algorithm is not applicable here. The focus is on the analytical and technical performance of the assay itself.
9. How the ground truth for the training set was established
As this is not an AI/ML device, there is no "training set" or corresponding ground truth establishment process for algorithm training. The ground truth for the various performance evaluation studies (e.g., confirming p16 expression in Western Blots, identifying known tissue types) would be established through standard laboratory and pathological methods.
§ 864.1865 Cervical intraepithelial neoplasia (CIN) test system.
(a)
Identification. A cervical intraepithelial neoplasia (CIN) test system is a device used to detect a biomarker associated with CIN in human tissues. The device is indicated as an adjunct test and not to be used as a stand-alone device. The test results must be interpreted in the context of the patient's clinical history including, but not limited to, prior and current cervical biopsy results, Papanicolaou (Pap) test results, human papillomavirus (HPV) test results, and morphology on hematoxylin and eosin (H&E) stained sections. This device is not intended to detect the presence of HPV.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include the following information:
(i) The indications for use must specify the biomarker that is intended to be identified and its adjunct use (
e.g., adjunct to examination of H&E stained slides) to improve consistency in the diagnosis of CIN.(ii) Summary of professional society recommendations, as applicable.
(iii) A detailed device description including:
(A) A detailed description of all test components, including all provided reagents and required, but not provided, ancillary reagents.
(B) A detailed description of instrumentation and equipment, including illustrations or photographs of non-standard equipment or manuals.
(C) If applicable, detailed documentation of the device software, including, but not limited to, stand-alone software applications and hardware-based devices that incorporate software.
(D) A detailed description of appropriate positive and negative controls that are recommended or provided.
(E) Detailed specifications for sample collection, processing, and storage.
(F) A detailed description of methodology and assay procedure.
(G) A description of the assay cutoff (the medical decision point between positive and negative) or other relevant criteria that distinguishes positive and negative results, including the rationale for the chosen cutoff or other relevant criteria and results supporting validation of the cutoff.
(H) Detailed specification of the criteria for test results interpretation and reporting.
(iv) Detailed information demonstrating the performance characteristics of the device, including:
(A) Analytical specificity studies such as, but not limited to, antibody characterization (
e.g., Western Blot, peptide inhibition analysis), studies conducted on panels of normal tissues and neoplastic tissues, interference by endogenous and exogenous substances as well as cross-reactivity, as applicable.(B) Device analytical sensitivity data generated by testing an adequate number of samples from individuals with the target condition including limit of blank, limit of detection, and limit of quantification, as applicable.
(C) Device precision/reproducibility data to evaluate within-run, between-run, between-day, between-lot, between-site, between-reader, within-reader and total precision, as applicable, using a panel of samples covering the device measuring range and/or the relevant disease categories (
e.g. No CIN, CIN1, CIN2, CIN3, cervical cancer) and testing in replicates across multiple, nonconsecutive days.(D) Device robustness/guardbanding studies to assess the tolerance ranges for various critical test and specimen parameters.
(E) Device stability data, including real-time stability and shipping stability under various storage times, temperatures, and freeze-thaw conditions.
(F) Data from a clinical study demonstrating clinical validity using well-characterized prospectively or retrospectively obtained clinical specimens, as appropriate, representative of the intended use population. The study must evaluate the consistency of the diagnosis of CIN, for example, by comparing the levels of agreements of diagnoses rendered by community pathologists to those rendered by a panel of expert pathologists. Agreement for each CIN diagnostic category (
e.g., No CIN, CIN1, CIN2, CIN3, cancer) and for alternate diagnostic categories (e.g., No CIN, low grade squamous intraepithelial lesion (LSIL)-histology, high grade squamous intraepithelial lesion (HSIL)-histology, cancer) between reference diagnosis by expert pathologist and community pathologist must be evaluated, as applicable. In addition, agreements for CIN binary categories as ≥CIN2 (i.e., CIN2 or CIN3 or cancer) and ≤CIN1 (i.e., No CIN or CIN1) between reference diagnosis by expert pathologist with H&E staining and community pathologist with H&E staining and agreements for alternate CIN binary categories as ≥HSIL-histology (i.e., HSIL-histology or cancer) and ≤LSIL-histology (i.e., No CIN or LSIL-histology) between reference diagnosis by an expert pathologist with H&E + [biomarker specified in paragraph (b)(1)(i) of this section] and a community pathologist with H&E + [biomarker specified in paragraph (b)(1)(i) of this section] must be evaluated and compared, as applicable.(G) The staining performance of the device as determined by the community pathologists during review of the study slides must be evaluated. The staining performance criteria assessed must include overall staining acceptability, background staining acceptability, and morphology acceptability, as applicable.
(H) Appropriate training requirements for users, including interpretation manual, as applicable.
(I) Identification of risk mitigation elements used by the device, including a description of all additional procedures, methods, and practices incorporated into the instructions for use that mitigate risks associated with testing.
(2) The device's 21 CFR 809.10(b) compliant labeling must include a detailed description of the protocol, including the information described in paragraph (b)(1)(ii) of this section, as applicable, and a detailed description of the performance studies performed and the summary of the results, including those that relate to paragraph (b)(1)(ii) of this section, as applicable.