(211 days)
The illumigene Mycoplasma DNA amplification assay, performed on the illumipro-10™, is a qualitative in vitro diagnostic test for the direct detection of DNA from Mycoplasma pneumoniae in human throat and nasopharyngeal swabs obtained from patients suspected of having Mycoplasma pneumoniae infection.
The illumigene Mycoplasma assay utilizes loop-mediated isothermal DNA amplification (LAMP) technology to detect Mycoplasma pneumoniae by targeting a segment of the Mycoplasma pneumoniae genome.
Results from the illumigene Mycoplasma DNA amplification assay should be used in conjunction with clinical presentation, other laboratory findings, and epidemiological risk factors as an aid in the diagnosis of Mycoplasma infection and should not be used as the sole basis for treatment or other patient management. Positive results do not rule out co-infection with other organisms and negative results in persons with respiratory tract infections may be due to pathogens not detected by this assay. Lower respiratory tract infections due to M. pneumoniae may not be detected by this assay. If lower respiratory tract infection due to M. pneumoniae is suspected, additional laboratory testing using methods other than the illumigene Mycoplasma DNA Amplification Assay may be necessary.
illumigene Mycoplasma is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.
The illumigene Molecular Diagnostic Test System is comprised of the illumigene® Mycoplasma DNA Amplification Assay Test Kit, the illumigene® Mycoplasma External Control Kit and the illumipro-10™ Automated Isothermal Amplification and Detection System.
The illumigene Mycoplasma assay utilizes loop-mediated isothermal amplification (LAMP) technology to detect the presence of Mycoplasma pneumoniae in human respiratory specimens (throat and nasopharyngeal swab specimens), Each illumigene Mvcoplasma assay is completed using an illumicene Assay Control Reagent containing Control material, an illumigene Reaction Buffer, an illumigene Mycoplasma Test Device and microcentrifuge tubes. Respiratory specimens are combined with the illumigene Assay Control Reagent. The Speciment is manually extracted and purfied using a commercially available extraction kit (Qiagen, QlAamp® DSP DNA Mini Kit). Extracted DNA is heat-treated. Target and Control DNA are made available for isothermal amplification via heattreatment. The heat-treated Specimen/Control sample is added to the illumigene Reaction Buffer. DNA amplification occurs in the illumigene Test Device.
The illumipro-10 heats each illumigene Mycoplasma Test Device containing prepared Sample and Control material. facilitating amplification of target DNA. When M. pneumoniae is present in the specimen, a 200 base pair sequence of the M. pneumoniae genome is amplified and magnesium pyrophosphate is generated. Magnesium pyrophosphate forms a precipitate in the reaction mixture.
The illumipro-10 monitors the absorbance characteristics of the reaction solutions at the assay Run Start (Signalinia) S) and at the assay Run End (Signalmar, S). The illumipro-10™ calculates the ratio of the Run End (Signal final or S) reads with the Run Start (Signal Initial or S) reads and compares the ratio to an established cut-off value. The illumipro-10 performs this ratio calculation to both the TEST chamber and the CONTROL chamber.
Fixed cut-off values for the CONTROL chamber are used to determine validity. CONTROL chamber S;S; ratios less than 90% are considered valid and allow for reporting of TEST chamber results (POSITIVE, NEGATIVE). CONTROL chamber S; S; ratios greater than or equal to 90% are considered invalid. Results are reported as 'INVALD': Test chamber results are not reported. More stringent cut-off criteria are applied to the Control chamber reaction to ensure amplification is not inhibited, reagents are performing as intended and that sample processing was performed appropriately.
Fixed cut-off values for the TEST chamber are used to report sample results. TEST chamber Sy:S ratios less than 82% are reported as 'POSITIVE'; TEST chamber S;S; ratios greater than or equal to 82% are reported as 'NEGATIVE'. Numerical values are not reported.
The illumigene Mycoplasma External Controls Kit contains a Positive Control Reagent, External Control reagents are provided to aid the user in detection of reagent deterioration, adverse environmental or test conditions, or variance in operator performance that may lead to test errors. External Control reagents are provided for use in routine Quality Control testing.
illumipro-10™ Automated Isothermal Amplification and Detection System:
The illumipro-10™ heats each illumigene Mycoplasma Test Device containing prepared samples and Control Reagent, facilitating amplification of target DNA. When Mvcoplasma pneumoniae is present in the respiratory swab sample, a conserved sequence of the M. pneumoniae is amplified and magnesium pyrophosphate is generated. Magnesium pyrophosphate forms a precipitate in the reaction mixture. The illumipro-10 detects the change in light transmission through the reaction mixture created by the precipitating magnesium pyrophosphate. The illumipro-10 reports sample results as INVALID, POSITIVE or NEGATIVE based on the detected change in light transmission.
The illumipro-10™ System Description was reviewed in previous submission. K10012. K112125. K121044 and K122019. No system or software changes were made for the illumigene Mycoplasma assay.
Here's a summary of the acceptance criteria and the study that proves the illumigene® Mycoplasma DNA Amplification Assay meets those criteria, based on the provided text:
Acceptance Criteria and Device Performance for illumigene® Mycoplasma DNA Amplification Assay
The illumigene® Mycoplasma DNA Amplification Assay is a qualitative in vitro diagnostic test for the direct detection of DNA from Mycoplasma pneumoniae in human throat and nasopharyngeal swabs. The performance was evaluated through clinical and non-clinical studies.
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined acceptance criteria in terms of numerical thresholds for sensitivity or specificity. Instead, it presents the achieved performance characteristics, which are then compared to a predicate device (FilmArray® Respiratory Panel (RP) System). The "acceptance criteria" are implied by the results needing to be comparable or superior to the predicate and generally indicative of accurate diagnostic performance.
Below is a summary of the device's reported clinical performance:
illumigene® Mycoplasma DNA Amplification Assay Performance
| Specimen Type | Specimen Description | Performance Metric | illumigene Performance | Predicate Device (FilmArray® RP) Performance |
|---|---|---|---|---|
| Nasopharyngeal Swab Specimens | ||||
| Prospective | % Sensitivity | 100.0% (95% CI: 51.0 - 100.0%) | 100.0% (95% CI: 39.8 – 100%) | |
| Prospective | % Specificity | 100.0% (95% CI: 92.6 - 100.0%) | 100.0% (95% CI: 99.7 - 100%) | |
| Retrospective | PPA (Positive Percent Agreement) | 94.4% (95% CI: 81.9 - 98.5%) | 84.4% (95% CI: 73.1 - 92.2%) | |
| Retrospective | NPA (Negative Percent Agreement) | 95.6% (95% CI: 89.1 - 98.3%) | 89.2% (95% CI: 79.1 - 95.6%) | |
| Throat Swab Specimens | ||||
| Prospective | % Sensitivity | 100.0% (95% CI: 67.6 - 100.0%) | Not Evaluated | |
| Prospective | % Specificity | 100.0% (95% CI: 91.8 - 100.0%) | Not Evaluated | |
| Retrospective | PPA | 84.6% (95% CI: 66.5 - 93.9%) | Not Evaluated | |
| Retrospective | NPA | 98.5% (95% CI: 92.0 - 99.7%) | Not Evaluated |
Note: The performance values presented are against a Composite Reference Method. For retrospective throat swabs, an additional comparison to PCR with Bi-Directional Sequencing is provided, showing PPA of 100.0% (95% CI: 84.5 - 100.0%) and NPA of 97.2% (95% CI: 90.4 - 99.2%).
2. Sample Size Used for the Test Set and Data Provenance
- Total Test Set Sample Size: 334 qualified throat and nasopharyngeal (NP) swab specimens.
- Split by Type:
- Prospective Samples: 103 specimens
- Retrospective Samples: 219 specimens
- Data Provenance:
- Country of Origin: Not explicitly stated, but the submission is to the US FDA by a US-based company (Meridian Bioscience), implying clinical studies were likely conducted in the US.
- Retrospective or Prospective: Both. The clinical study included both "leftover deidentified specimens submitted to the testing laboratories for routine M. pneumoniae testing. Specimens included in performance evaluation were prospective (never frozen) and retrospective (frozen prior to illumigene testing)."
3. Number of Experts Used to Establish Ground Truth for the Test Set and Their Qualifications
The document does not mention the number or specific qualifications of experts for establishing ground truth. The ground truth was established using a "Composite Reference Method."
4. Adjudication Method for the Test Set
The adjudication method relies on a "Composite Reference Method" as the ground truth. This method included:
- M. pneumoniae bacterial culture with identification
- A validated real-time PCR assay followed by bi-directional sequencing.
Specimens were considered positive if they produced positive Mycoplasma pneumoniae results from either bacterial culture or real-time PCR and bi-directional sequencing. Specimens negative for both culture and PCR were considered negative. No mention of an explicit expert adjudication process for discordant results is made beyond the composite reference method itself.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was mentioned. The device is an automated DNA amplification assay, not an imaging device requiring human reader interpretation. Therefore, a study to assess human reader improvement with AI assistance is not applicable in this context.
6. Standalone Performance Study (Algorithm Only)
Yes, a standalone performance study was conducted. The "illumigene® Mycoplasma DNA Amplification Assay, performed on the illumipro-10™" is an automated device designed to produce results (POSITIVE, NEGATIVE, INVALID) directly. The performance data presented in the tables (sensitivity, specificity, PPA, NPA) reflect the standalone performance of the illumigene system.
7. Type of Ground Truth Used
The ground truth used was a Composite Reference Method which included:
- M. pneumoniae bacterial culture with identification.
- A validated real-time PCR assay followed by bi-directional sequencing.
- Positive: A specimen was considered positive if either bacterial culture or real-time PCR and bi-directional sequencing yielded a positive result.
- Negative: A specimen was considered negative if both culture and PCR were negative.
8. Sample Size for the Training Set
The document does not explicitly specify a "training set" for the clinical performance evaluation. The clinical studies evaluated the performance of the already developed assay on clinical specimens. The assay's internal "optimization" during development would have involved some form of training/tuning, but the size of data used for this is not detailed in the summary. For non-clinical performance (e.g., precision, LoD), contrived samples were used for development and internal testing.
9. How the Ground Truth for the Training Set Was Established
For the clinical study used to demonstrate performance: The ground truth was established by the Composite Reference Method as described in point 7.
For the development/optimization of the assay: The document states that "Development optimization includes evaluation of characterized positive clinical specimens. Amplification reagent concentrations are adjusted during design as needed to ensure illumigene results are aligned with clinical specimen reported results." This implies that well-characterized clinical specimens and potentially cultured strains were used to refine the assay's parameters and establish cut-off values. However, details on the specific samples or methods for establishing ground truth for this development/training phase are not provided.
§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.
(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.