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2	Genetic and Molecular Tests Currently, only ~ 1% of genetic tests available to clinicians are FDA cleared or approved FDA is concerned that commercially distributed genetic tests are reliable and that patients and health care professionals understand both the value and the limitations of such testing
3	“Instrumentation for Clinical Multiplex Test Systems” Intended to measure and sort multiple signals generated by an assay from a clinical sample Instrumentation is used with a specific assay to measure multiple similar analytes that establish a single indicator to aid in diagnosis Device includes: signal reader unit, → raw data storage mechanisms integrated reagent handling, → data acquisition hybridization, washing, → software to process signal dedicated instrument control, other hardware components.
4	“Instrumentation for Clinical Multiplex Test Systems” (cont. I) Guidance addresses multiplex tests that measure multiple similar analytes that establish a single indicator to aid in diagnosis: SNPs patient’s genotype multiple alleles Instrumentation could aid in clinical applications: screening or diagnosis of disease drug selection and dosing patient management assessment of disease progression and regression
5	“Instrumentation for Clinical Multiplex Test Systems” (cont. II) A specific multiplex assay or test that is intended to be run on instrumentation for clinical multiplex test systems is considered a separate device This guidance document does not address performance characteristics for such an assay
6	“Instrumentation for Clinical Multiplex Test Systems” (cont. III) Risk to Health Inaccurate or absent results due to failure of instrumentation components (sections 7, 9) Inaccurate results due to failure of data management and database software (section 8)
7	“Instrumentation for Clinical Multiplex Test Systems” (cont. IV) Intended Use Claims in the intended use determine the type of review that is necessary - (often independent of the technology or assay format) Separate submissions for multiple intended uses (e.g., a device that genotypes multiple genes) Carefully define the intended use population(s) (e.g., some alleles/genotypes may have low prevalence in certain pops - many patients may need to be tested for statistical significance)
8	“Instrumentation for Clinical Multiplex Test Systems” (cont. V) Device Description Describe additional instrument components provided, or recommended for use, and their function in the system How the instrument is designed to carry out its functions related to amplification, hybridization, or signal detection The sample type(s) that may be run on the instrumentation Types of output generated by the instrumentation and system parameters (e.g., reading ranges) Related peer-reviewed literature references (if applicable)
9	“Instrumentation for Clinical Multiplex Test Systems” (cont. VI) Analytical performance Precision (reproducibility) characterize reproducibility using well-characterized sample + positive and negative control assays assess overall instrumentation performance justify statistically sample size for reproducibility study ≥ 2 operators and ≥ 3 instruments in the study masked samples (results unknown to operators) report between-assay, between-scan, between-instrument, between-operator, or other evaluations, as appropriate
10	“Instrumentation for Clinical Multiplex Test Systems” (cont. VII) Analytical performance Recommend to report: Coefficients of variation (CV) with CI for between- instruments, operators, device lots, and other evaluations, as appropriate Pairwise correlation coefficients, scatter plots, and ANOVA analysis of data from all relevant elements of the reproducibility study Any bias observe during reproducibility studies and explanation to account for observed bias
11	“Instrumentation for Clinical Multiplex Test Systems” (cont. VIII) Analytical performance Method Comparison: evaluate performance of your instrumentation against predicate device ( % agreement) NO predicate available compare performance to “gold standard” method Calibration: develop and recommend calibration program/maintenance schedule for each portion of the instrumentation
12	“Instrumentation for Clinical Multiplex Test Systems” (cont. IX) Software Fully describe software design Do not include software utilities that support uses beyond your assay consider privacy and security issues in your design. Submit hazard analysis based on impact of any failure of subsystem components signal detection and analysis, data storage, system communications and cybersecurity Complete verification and validation (V&V) activities
13	“Drug Metabolizing Enzyme Genotyping System” Intended to test DNA to identify presence/absence of human genotypic markers encoding DME Device is used an aid to determine: treatment choice individualizing treatment dose for therapeutics
14	“Drug Metabolizing Enzyme Genotyping System” (cont. I) Risk to Health Failure to correctly identify genotype encoding for a DME (sections 7,8, and 9) Failure to properly interpret genotyping results (section 10)
15	“Drug Metabolizing Enzyme Genotyping System” (cont. II) Intended Use Specify genotype(s) the test is intended to detect General clinical utility of detecting the genotype Specific populations to which the test is targeted Specify the drug metabolizing enzyme encoded by the genotype (tests with multiple intended uses (e.g., multiple DME’s): When separate studies are needed to support the multiple intended uses, submit separate 510(k) applications for each intended use)
16	“Drug Metabolizing Enzyme Genotyping System” (cont. III) Performance Characteristics Description of samples used in testing (including types of samples, preparation or origin, number of samples and how the samples specifically represent your intended use samples). Descriptions of the statistical methods you used. Explanations, if there were any deviations from your protocol.
17	“Drug Metabolizing Enzyme Genotyping System” (cont. IV) Performance Characteristics Preanalytical Factors Critical for high-quality genetic tests If extraction and preparation of DNA for testing is provided (validate each step, reproducibility) Not DNA reagents provided -> present specifications for quality of the assay input DNA
18	“Drug Metabolizing Enzyme Genotyping System” (cont. V) Performance Characteristics Quality Control Include both positive and negative controls (controls should reflect sample composition/DNA concentration Controls should show that all steps/critical reactions have proceeded properly and without contamination or cross-hybridization
19	“Drug Metabolizing Enzyme Genotyping System” (cont. VI) Analytical performance Precision (reproducibility) Accuracy Limit of Detection Traceability Potential Interferences
20	“Drug Metabolizing Enzyme Genotyping System” (cont. VII) Analytical performance Precision (Reproducibility) (EP5-A/EP12-A) characterize intra- and inter-assay reproducibility samples used in reproducibility testing are processed from “real” samples (e.g. whole blood, buccal swabs, or others) at test site (processing should mimics procedure recommended in the test) ≥3 sites with multiple operators at each site. Operators should reflect potential users of the assay (education and experience) multiple product lots, and multiple instruments (if instruments are part of the test system)
21	“Drug Metabolizing Enzyme Genotyping System” (cont. VIII) Accuracy:
22	“Drug Metabolizing Enzyme Genotyping System” (cont. IX) Limit of Detection Lowest (and highest) concentration of input sample that yields a reliable and accurate result Potential Interferences Endogenous and exogenous common substances Commonly prescribed drugs Similar molecules Sample preparation / conditions Effect of excess sample or limiting sample
23	“Drug Metabolizing Enzyme Genotyping System” (cont. X) Software: Total system validation Describe any algorithms or cutoffs used to determine results Database integrity Cyber-security Follow the FDA guidance for premarket submission requirements for devices containing software Follow the FDA guidance for off-the-shelf software
24	“Drug Metabolizing Enzyme Genotyping System” (cont. XI) Clinical performance – clinical validity Device must have a clinical indication/clinical utility May be based on: Existing clinical data New clinical trial data Review of information in the literature Current clinical knowledge
25	“Drug Metabolizing Enzyme Genotyping System” (cont. XII) When clinical studies are needed to show clinical utility: Prospective clinical studies Retrospective validation Banked samples from prior clinical studies Samples should be stored under optimal conditions Bridging studies may be done if a platform change or device change is necessary after clinical validation
26	GOOD SCIENCE