The MycoMEIA® Aspergillus Assay (MycoMEIA) is an enzyme immunoassay (EIA) for the in vitro qualitative detection of Aspergillus antigens in human urine from adults (>/= 18 years old) with suspected invasive aspergillosis (IA). The assay is not intended for use in lung transplant recipients. The results should be interpreted by trained healthcare professionals, incorporating other diagnostic procedures such as microbiological culture, histological examination of biopsy samples, and radiographic evidence to support the diagnosis of IA.

Device Description

The MycoMEIA® Aspergillus Assay (MycoMEIA) is a sandwich enzyme immunoassay (EIA) that detects Aspergillus antigens in urine. The assay uses two mouse monoclonal antibodies that recognize galactofuranose (galf) epitopes. Both monoclonal antibodies are used to coat the wells of the microplate to bind the antigen, and to detect the antigen bound to the sensitized microplate as horseradish-peroxidase conjugates.

The design of the assay is in 96-well plates to enable large-volume testing in clinical laboratories.

Urine is first processed through Sample Processing Columns to eliminate an inhibitor of antibody-antigen recognition. The processed urine samples are added to the plate wells coated with the antibodies, incubated, and washed to remove unbound material. The bound antigen is incubated with antibodies linked to horseradish peroxidase and washed to remove the unbound material.

Next, the peroxidase substrate solution is added, which reacts with the complexes bound to the well to form a blue color reaction. The enzyme reaction is stopped by the addition of acid, which changes the blue color to yellow. The absorbance (optical density) of specimens and controls is determined with a spectrophotometer set at 450 and 620 nm wavelengths.

The amount of antigen in the clinical sample is determined by optical density (OD) using a spectrophotometer and interpreted as an OD index relative to the mean OD of a threshold control provided in the kit.

The MycoMEIA-ASP kit contains the following components:

Microwell Plate
Negative Control (NC) Sample (green)
Threshold Control (TC) Sample (blue)
Positive Control (PC) Sample (red)
Conjugate (100X) (white)
Conjugate Diluent (white)
Chromogen Solution (yellow)
Stop Solution (blue)
Column Rinse
Plate sealers

25X Concentrated Wash Solution and Sample Processing columns are required to perform the assay and are provided separately from the kit.

If a partial plate is used, empty ELISA plates to fill in the ELISA frame are available to customers as a separate product.

Each laboratory will be required to test positive and negative controls provided in the kit to confirm or authenticate the assay results and to determine the sample index factor that is calculated using the assay ODs. Calculations can be performed manually or using the WS001 MycoMEIA Calculation Worksheet, which is available separately.

AI/ML Overview

The provided FDA 510(k) clearance letter and summary for the MycoMEIA Aspergillus Assay details the device's analytical and clinical performance. Here's a breakdown of the acceptance criteria and study information:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state formal "acceptance criteria" for clinical performance in a pass/fail manner. Instead, it presents the determined performance characteristics. For several analytical aspects, the criteria are implicitly defined by standard guidelines (e.g., CLSI EP17-A2 for LoD, CLSI EP05-A3 for precision and reproducibility).

Acceptance Criteria Category	Specific Metric (Implicit or Explicit)	Reported Device Performance
Analytical Sensitivity	Limit of Detection (LoD)	3 ng/mL
Precision (Repeatability)	MycoMEIA Positive Control OD CV	5%
	MycoMEIA Positive Control IDX CV	5%
Precision (Within-Lab)	MycoMEIA Positive Control OD CV	9%
	MycoMEIA Positive Control IDX CV	10%
Reproducibility (Overall)	MycoMEIA Positive Control OD CV	13%
	MycoMEIA Positive Control Index CV	13%
Interfering Substances	No interference at defined levels	Observed cross-reactivity with 10% v/v Plasma-Lyte A and certain high concentrations of other substances.
Cross-Reactivity	No cross-reactivity with specified organisms	Observed cross-reactivity with Fusarium (8.80 x 10e5 CFU/mL). Also detected from clinical samples with histoplasmosis, blastomycosis, candidemia, streptococcus, rhinovirus/parainfluenzavirus, mixed bacterial infection, mixed GI complications, and disseminated fusariosis.
Clinical Sensitivity (Per-Subject)	For proven & probable IA	92.4% (95% CI 82.1-97.0%)
Clinical Sensitivity (Per-Sample)	For proven & probable IA	58.2% (95% CI 54.3-61.9%)
Clinical Specificity (Prospective Study)	No IA, using Positive cutoff ≥ 0.6	86.1% (95% CI 75.7-92.5%)
High-Dose Hook Effect	Absence of hook effect	No high-dose hook effect observed up to 1,000 ng/mL.

2. Sample Size Used for the Test Set and Data Provenance

Analytical Sensitivity (LoD): 60 determinations with low target analyte level replicates.
Precision (Repeatability and Within-Laboratory): 80 replicates per sample (run twice daily for 20 days, by two operators, in duplicate). Samples included kit controls, contrived samples, and clinical sample pools.
Reproducibility: 90 replicates per sample (run twice daily for 5 days, by two operators, in triplicate) across three sites. Samples included kit controls, contrived samples, and clinical sample pools.
Interfering Substances: Endogenous and exogenous substances spiked into pooled healthy urine, and clinical samples from patients with known endogenous conditions.
Cross-Reactivity (Microorganisms): Tested in duplicate (n=2) for each microorganism listed.
Cross-Reactivity (Clinical Samples): Urine samples from people with various medical conditions, including fungal, viral, and bacterial infections.
- Specific numbers for each condition are listed in the summary (e.g., 1 with histoplasmosis, 2 with streptococcus, etc.).
- Additional cross-reactivity testing involved 21 subjects with documented bacterial pneumonia, 1 with mixed bacterial infection, 2 with mixed GI complications, 2 with histoplasmosis, 1 with blastomycosis, 1 with candidemia, and 1 with disseminated fusariosis.
Clinical Performance:
- Archived, Retrospective Study: 475 samples collected from 290 subjects. 226 samples from 50 subjects with proven (n=3) or probable (n=47) IA.
- Prospective Study:
  - Initiated with 210 consented subjects providing 254 urine samples during 213 suspected infection episodes.
  - Excluded 34 lung transplant recipients, 2 pediatric subjects, 1 contaminated sample, 3 subjects without CT scans, and 4 subjects who received mold-active antifungal therapy.
  - Final analysis: 166 subjects and 169 infectious episodes for specificity.
  - "Possible" IA cases (n=106) were excluded from the analysis for specificity.
  - 38 subjects were adjudicated as having no invasive fungal infection.
  - 26 subjects were adjudicated as having mixed or other (non-IA) infections.

Data Provenance:

Clinical Performance: Comprised of both archived, retrospective samples and prospectively collected samples.
Cross-Reactivity, Interfering Substances, Analytical Sensitivity, Precision, Reproducibility: Laboratory-based studies using spiked samples, controls, and some clinical samples (origin of clinical samples not explicitly stated beyond "healthy urine" or "clinical samples from patients with known endogenous conditions" or "urine samples obtained from people with different medical conditions").

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

Clinical Performance (Prospective Study): Clinical diagnoses were adjudicated by "a reviewer who was blinded to MycoMEIA test results."
Qualifications: The specific qualifications of this reviewer (e.g., specialty, years of experience) are not specified in the provided text.

4. Adjudication Method for the Test Set

Clinical Performance (Prospective Study): The adjudication method involved "a reviewer who was blinded to MycoMEIA test results." The reviewer applied the 2020 EORTC/MSG diagnostic criteria to establish diagnoses of proven IA, probable IA, and possible IA.
No specific multi-expert adjudication method (e.g., 2+1, 3+1) is mentioned.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size of Human Improvement with AI vs. without AI Assistance

The MycoMEIA Aspergillus Assay is an enzyme immunoassay (EIA), an in vitro diagnostic (IVD) device for detecting antigens.
This is not an AI-powered device or imaging-based device that would involve human readers interpreting images with or without AI assistance.
Therefore, an MRMC comparative effectiveness study involving human readers and AI assistance was not conducted, and such data is not applicable here.

6. If a Standalone (i.e., Algorithm Only Without Human-in-the-Loop Performance) Was Done

This is an in vitro diagnostic assay, which intrinsically provides a standalone (assay-only) result (an index value). The device itself is the algorithm/assay.
The clinical performance section (sensitivity and specificity tables) represents the standalone performance of the MycoMEIA assay in detecting Aspergillus antigens in urine.
While results are "interpreted by trained healthcare professionals, incorporating other diagnostic procedures," the reported sensitivity and specificity values reflect the assay's direct performance against the established ground truth.

7. The Type of Ground Truth Used

Clinical Performance: The ground truth for Invasive Aspergillosis (IA) was established using 2020 EORTC/MSG diagnostic criteria for proven and probable IA. This is a consensus-based diagnostic standard.
Analytical Studies (LoD, Precision, Reproducibility, Interfering Substances, Cross-Reactivity): Ground truth was based on known concentrations of Aspergillus antigen, specific microorganisms, or specific interfering substances as used in controlled spiking experiments.

8. The Sample Size for the Training Set

The provided document describes a 510(k) submission for a clinical assay. It details analytical and clinical performance studies, which serve as validation.
Unlike machine learning or AI-based devices, traditional IVDs typically do not involve a distinct "training set" in the same sense. The assay's design and optimization (analogous to training) would have occurred during its development phase, but specific "training set" sample sizes are not applicable or provided in this context. The study data presented are for validation/testing.

9. How the Ground Truth for the Training Set Was Established

As explained above, a distinct "training set" with established ground truth in the context of machine learning is not relevant for this traditional immunoassay device. The ground truth for validation data was established as per point 7.

Summary

FDA 510(k) Clearance Letter - MycoMEIA Aspergillus Assay

Page 1

U.S. Food & Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993
www.fda.gov

Doc ID # 04017.08.00

August 1, 2025

Pearl Diagnostics, Inc.
Irina Baburina
VP, R&D and Regulatory
1812 Ashland Ave
Ste G32
Baltimore, Maryland 21205

Re: K243496
Trade/Device Name: MycoMEIA Aspergillus Assay
Regulation Number: 21 CFR 866.3040
Regulation Name: Aspergillus Spp. Serological Reagents
Regulatory Class: Class I
Product Code: NOM
Dated: July 3, 2025
Received: July 3, 2025

Dear Irina Baburina:

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 (the 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 available 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.

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K243496 - Irina Baburina Page 2

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-devices/device-advice-comprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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-devices/medical-device-safety/medical-device-reporting-mdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medical-devices/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-devices/device-advice-comprehensive-regulatory-

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K243496 - Irina Baburina Page 3

assistance/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,

Ribhi Shawar -S

Ribhi Shawar, Ph.D. (ABMM)
Branch Chief, General Bacteriology and Antimicrobial Susceptibility Branch
Division of Microbiology Devices
OHT7: Office of In Vitro Diagnostics
Office of Product Evaluation and Quality
Center for Devices and Radiological Health

Enclosure

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FORM FDA 3881 (8/23) Page 1 of 1 PSC Publishing Services (301) 443-6740 EF

DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration

Indications for Use

Form Approved: OMB No. 0910-0120
Expiration Date: 07/31/2026
See PRA Statement below.

510(k) Number (if known): K243496

Device Name: MycoMEIA Aspergillus Assay

Indications for Use (Describe):

The MycoMEIA Aspergillus Assay (MycoMEIA) is an enzyme immunoassay (EIA) for the in vitro qualitative detection of Aspergillus antigens in human urine from adults (>/= 18 years old) with suspected invasive aspergillosis (IA). The assay is not intended for use in lung transplant recipients. The results should be interpreted by trained healthcare professionals, incorporating other diagnostic procedures such as microbiological culture, histological examination of biopsy samples, and radiographic evidence to support the diagnosis of IA.

Type of Use (Select one or both, as applicable):
☒ Prescription Use (Part 21 CFR 801 Subpart D)
☐ Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services
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"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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510(k) Summary

510(k) #: K243496
Date prepared: 01 August 2025

Applicant: Pearl Diagnostics, Inc.
1812 Ashland Ave Ste G32
Baltimore MD 21205
Tel: +1-317-413-3465

Contact: Irina Baburina

Device Trade Name: MycoMEIA® Aspergillus Assay
Common Name: Aspergillus spp. serological reagents
Classification Name: Antigen, Galactomannan, Aspergillus Spp.
Regulation Number: 866.3040
Product Code(s): NOM

Predicate Trade Name: Bio-Rad Laboratories Platelia™ Aspergillus EIA
Predicate 510(k) #: K093678
Predicate Product Code: NOM

Device Description Summary:

The design of the assay is in 96-well plates to enable large-volume testing in clinical laboratories.

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bound antigen is incubated with antibodies linked to horseradish peroxidase and washed to remove the unbound material.

The MycoMEIA-ASP kit contains the following components:

Microwell Plate
Negative Control (NC) Sample (green)
Threshold Control (TC) Sample (blue)
Positive Control (PC) Sample (red)
Conjugate (100X) (white)
Conjugate Diluent (white)
Chromogen Solution (yellow)
Stop Solution (blue)
Column Rinse
Plate sealers

25X Concentrated Wash Solution and Sample Processing columns are required to perform the assay and are provided separately from the kit.

If a partial plate is used, empty ELISA plates to fill in the ELISA frame are available to customers as a separate product.

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Intended Use/Indications for Use:

The MycoMEIA® Aspergillus Assay (MycoMEIA) is an enzyme immunoassay (EIA) for the in vitro qualitative detection of Aspergillus antigens in human urine from adults (≥18 years old) with suspected invasive aspergillosis (IA). The assay is not intended for use in lung transplant recipients.

The results should be interpreted by trained healthcare professionals, incorporating other diagnostic procedures such as microbiological culture, histological examination of biopsy samples, and radiographic evidence to support the diagnosis of IA.

Rx only

For in vitro diagnostic use

Comparison to Predicate:

Characteristic	Predicate Device Bio-Rad Laboratories Platelia™ Aspergillus EIA K093678	Subject Device MycoMEIA® Aspergillus Assay (MycoMEIA)
Intended Use/Indications for Use	The Platelia Aspergillus Ag EIA is an immunoenzymatic sandwich microplate assay for the detection of Aspergillus galactomannan antigen in adult and pediatric serum and Bronchoalveolar Lavage (BAL) fluid samples. The Platelia Aspergillus EIA is a test which, when used in conjunction with other diagnostic procedures such as microbiological culture, histological examination of biopsy samples and radiographic evidence, can be used as an aid in the diagnosis of Invasive Aspergillosis.	The MycoMEIA® Aspergillus Assay (MycoMEIA) is an enzyme immunoassay (EIA) for the in vitro qualitative detection of Aspergillus antigens in human urine from adults (>/= 18 years old) with suspected invasive aspergillosis (IA). The assay is not intended for use in lung transplant recipients. The results should be interpreted by trained healthcare professionals, incorporating other diagnostic procedures such as microbiological culture, histological examination of biopsy samples, and radiographic evidence to support the diagnosis of IA.
Analyte	Aspergillus galactomannan	Aspergillus galf-containing antigens
Sample Type	Serum, BAL fluid	Urine
Technology	Enzyme immunoassay	Enzyme immunoassay

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Characteristic	Predicate Device Bio-Rad Laboratories Platelia™ Aspergillus EIA K093678	Subject Device MycoMEIA® Aspergillus Assay (MycoMEIA)
Reagent Components	1. 96-well plate coated with mAbEBA22. Labeled mAb containing preservative	1. 96-well plate coated with mAb476 and mAb5012. Labeled mAb containing preservative
Required Platform	Microplate reader	Microplate reader
Testing Environment	Clinical Laboratory	Clinical Laboratory
Reagent Condition and Storage	Liquid, 2-8°C	Liquid, 2-8°C
Sample Preparation	EDTA pretreatment boil at 120°C and centrifugation	Sample elution through Sample Processing Columns
Incubation Time	90 ± 5 minutes	Step 1 (sample addition) - 60+/- 2 min Step 2 (conjugate addition) - 60+/- 2 min
Incubation Temperature	37°C	37°C
Test Output	Index value calculated by dividing sample OD by Cut-off control OD. Positive results I ≥ 0.5.	Index values calculated by dividing by TC OD and multiplying by multiplication factor. Positive (≥ 0.6), Negative (< 0.6). Repeat testing is recommended for index values of 0.6 to 0.7.

Non-Clinical and/or Clinical Tests Summary & Conclusions:

The following nonclinical/analytical studies are submitted in this 510(k) for the determination of substantial equivalence:

ANALYTICAL SENSITIVITY

The LoD is the lowest amount of analyte that can be reliably detected (≥ 95% of results are clinically interpreted as "Positive"). The LoD of the assay is 3 ng/mL. The LoD was determined to be consistent with the guidelines in CLSI document EP17-A2 based on 60 determinations with low target analyte level replicates.

PRECISION

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Within-lab precision and repeatability were determined according to CLSI Guideline EP05-A3. Samples included kit controls (NC, TC, PC), contrived samples with known Aspergillus antigen concentrations (negative, low positive, positive), and clinical sample pools. Assays were run twice daily (AM and PM) using three kit lots by two different operators for twenty days. Each sample was tested in duplicate (n=2) for each run for a total N of 80 replicates per sample. Mean OD and index (IDX) values, standard deviation (SD), % Coefficient of variation (%CV) were calculated to determine repeatability (intra-assay variation) and within-laboratory precision (inter-assay variation). The coefficient variation results are shown for positive samples only.

Results:

MycoMEIA Positive Control - Mean OD = 1.909, Mean IDX = 22.6
Repeatability: OD SD = 0.090, OD CV = 5%, IDX SD = 1.076, IDX CV = 5%
Within-Laboratory Precision: OD SD = 0.163, OD CV = 9%, IDX SD = 2.174, IDX CV = 10%

MycoMEIA Threshold Control - Mean OD = 0.603, Mean IDX = 7.1
Repeatability: OD SD = 0.048, OD CV = 8%, IDX SD = 0.587, IDX CV = 8%
Within-Laboratory Precision: OD SD = 0.080, OD CV = 13%, IDX SD = 0.915, IDX CV = 13%

MycoMEIA Negative Control - Mean OD = 0.019, Mean IDX = 0.2
Repeatability: OD SD = 0.002, OD CV = NA, IDX SD = 0.023, IDX CV = NA
Within-Laboratory Precision: OD SD = 0.004, OD CV = NA, IDX SD = 0.047, IDX CV = NA

Negative Pool - Mean OD = 0.022, Mean IDX = 0.3
Repeatability: OD SD = 0.002, OD CV = NA, IDX SD = 0.023, IDX CV = NA
Within-Laboratory Precision: OD SD = 0.003, OD CV = NA, IDX SD = 0.038, IDX CV = NA

Contrived Low Positive Sample - Mean OD = 0.065, Mean IDX = 0.8

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Repeatability: OD SD = 0.007, OD CV = 10%, IDX SD = 0.077, IDX CV = 10%
Within-Laboratory Precision: OD SD = 0.009, OD CV = 13%, IDX SD = 0.106, IDX CV = 14%

Contrived Positive Sample - Mean OD = 0.118, Mean IDX = 1.4
Repeatability: OD SD = 0.006, OD CV = 5%, IDX SD = 0.069, IDX CV = 5%
Within-Laboratory Precision: OD SD = 0.013, OD CV = 11%, IDX SD = 0.137, IDX CV = 10%

Clinical Sample Pool - Mean OD = 0.135, Mean IDX = 1.6
Repeatability: OD SD = 0.0008, OD CV = 6%, IDX SD = 0.094, IDX CV = 6%
Within-Laboratory Precision: OD SD = 0.0133, OD CV = 10%, IDX SD = 0.171, IDX CV = 11%

Clinical Sample Pool - Mean OD = 0.238, Mean IDX = 2.8
Repeatability: OD SD = 0.010, OD CV = 4%, IDX SD = 0.112, IDX CV = 4%
Within-Laboratory Precision: OD SD = 0.038, OD CV = 16%, IDX SD = 0.515, IDX CV = 18%

REPRODUCIBILITY

Reproducibility was determined according to CLSI Guideline EP05-A3. Samples included kit controls (NC, TC, PC), contrived samples with known Aspergillus antigen concentrations (negative, low positive, positive), and clinical sample pools. Assays were run twice daily (AM and PM) using one kit lot by two different operators for five days. Each sample was tested in triplicate (n=3) for each run for a total N of 90 for each sample. Assays were run at three sites. Mean OD and index (IDX) values, standard deviation (SD), % Coefficient of variation (%CV) were calculated to determine repeatability, within-laboratory precision, and reproducibility. Coefficient of variation results are shown for positive samples only.

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MycoMEIA Results for All Sites:

MycoMEIA Positive Control - Mean OD = 1.580, SD = 0.213, CV = 13%
MycoMEIA Positive Control - Mean Index = 19.0, SD = 2.470, CV = 13%

MycoMEIA Threshold Control - Mean OD = 0.515, SD = 0.104, CV = 20%
MycoMEIA Threshold Control - Mean Index = 6.2, SD = 1.077, CV = 17%

MycoMEIA Negative Control - Mean OD = 0.024, SD = 0.011, CV = NA
MycoMEIA Negative Control - Mean Index = 0.3, SD = 0.123, CV = NA

Negative Human Urine Pool - Mean OD = 0.031, SD = 0.012, CV = NA
Negative Human Urine Pool - Mean Index = 0.4, SD = 0.138, CV = NA

Contrived Low Positive Sample - Mean OD = 0.080, SD = 0.019, CV = 24%
Contrived Low Positive Sample - Mean Index = 1.0, SD = 0.210, CV = 22%

Contrived Positive Sample - Mean OD = 0.142, SD = 0.031, CV = 22%
Contrived Positive Sample - Mean Index = 1.7, SD = 0.318, CV = 19%

Clinical Sample Pool - Mean OD = 0.084, SD = 0.016, CV = 19%
Clinical Sample Pool - Mean Index = 1.0, SD = 0.186, CV = 18%

Clinical Sample Pool - Mean OD = 0.207, SD = 0.029, CV = 14%
Clinical Sample Pool - Mean Index = 2.5, SD = 0.297, CV = 12%

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INTERFERING SUBSTANCES

Interference of the assay was determined according to CLSI Guideline EP07. Interference was tested using endogenous and exogenous substances spiked into pooled healthy urine, and by testing clinical samples from patients with known endogenous conditions, as indicated by abnormal urinalysis results. Interferents and conditions tested are shown in the table below. Contrived samples were prepared with 8 ng/mL antigen, approximately 3X the assay LOD. OD was tested before and after addition of Aspergillus antigen to assess potential cross-reactivity caused by the exogenous or the endogenous substances.

Among the substances and conditions tested, cross-reactivity was observed with 10% v/v Plasma-Lyte A.

Interference details are provided in the Limitations of the Procedure section of the Instructions for Use. Cross-reactivity was observed with negative samples, resulting in false positive results in the absence of Aspergillus antigen.

Results of testing contrived samples with endogenous and exogenous materials:

No interference was observed in samples with the following:

1-Methylnicotinamide (Vitamin B3 metabolite) 0.130 mg/mL
Acetaminophen 2.5 mg/mL
Acetone (Ketone) 2.4 mg/mL
Albumin 10 mg/mL
Amoxicillin 1.59 mg/mL
Amphotericin B 0.22 mg/mL
Ascorbic acid (Vitamin C) 0.5 mg/mL, 1 mg/mL
Azithromycin 0.15 mg/mL
Biotin (Vitamin B) 0.4 µg/mL
Boric Acid 7.89 mg/mL
CD14 3 µg/mL
Cimetidine 0.9 mg/mL
Ciprofloxacin 1.2 mg/mL

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Erythromycin 0.675 mg/mL
Ethanol 30 mg/mL
Ganciclovir hydrate 0.9 mg/mL
Glucose 6 mg/mL
Human chorionic gonadotropin (HCG) 0.3 µg/mL
Ibuprofen 0.27 mg/mL
Immunoglobulin G 2 mg/mL
Itraconazole 0.22 mg/mL
Metronidazole 1 mg/mL
Mucin 5% w/v
Mucin-5B 0.1% w/v
Naproxen sodium 0.02 mg/mL
Nicotine 0.015 mg/mL
Orosomucoid 0.2 µg/mL
Oxalic acid 1 mg/mL, 0.1 mg/mL
Penicillin G sodium salt 3.2 mg/mL
Phenazopyridine HCl 1.2 µg/mL
Phylloquinone (Vitamin K1) 0.05 µg/mL
Piperacillin sodium salt 19 mg/mL
Potassium clavulanate 0.65 mg/mL
RBC (erythrocytes) 2% v/v
Riboflavin (Vitamin B2) 0.24 mg/mL
Rifampicin 10 mg/mL
Sodium salicylate 4.73 mg/mL
Tazobactam sodium salt 17 mg/mL
Triglyceride 1 mg/mL, 0.3 mg/mL

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Urobilinogen 0.06 mg/mL
Vancomycin HCl 3.1 mg/mL
Voriconazole N-oxide 0.022 mg/mL
Vanillylmandelic acid 0.03 mg/mL
Water-based personal lubricant 50 mg/mL
WBC (White Blood Cells / Leukocytes) 300,000 cells/mL
α1-Acid glycoprotein 0.6 µg/mL
α-CEHC (metabolite of Vitamin E) 5 µg/mL
β-carotene (parent compound of Vitamin A) 3 mg/mL
Contrived alkaline pH 10.0
Contrived acidic pH 3.82
Abnormal clinical urine (Protein 30 mg/dL, pH 8.0, Hemolyzed blood 200 Ery/µL, Ketones >160mg/dL, Bilirubin +++, Glucose 250mg/dL)
Abnormal clinical urine (pH 7.51, Bilirubin +++)
Abnormal clinical urine (Ketones 80 mg/dL, Bilirubin ++, Glucose 250 mg/dL)
Acidic pH (pH 5.16)
High protein 100 mg/dL
Normal clinical urine
Normal clinical urine
Betanin 0.05 mg/mL
Bilirubin 0.03 mg/mL
Caffeine (in urine) 0.16 mg/mL
Gyne-Lotrimin cream (clotrimazole) 0.14% w/v
Hemoglobin 0.15 mg/mL
Lotrimin Ultra cream (butenafine) 0.14% w/v
Monistat 7 cream (miconazole) 0.14% w/v

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Nitrites 0.04 mg/mL
Vaginal contraceptive gel 0.14% w/v
Vagisil cream (benzocaine) 0.14% w/v
Vagistat 1 cream (tioconazole) 0.14% w/v

Cross-reactivity was observed at high concentrations in samples with the following:

Betanin 0.25% w/v
Bilirubin 0.06 mg/mL
Caffeine 6.75 mg/mL
Gyne-Lotrimin cream 2.5% w/v
Hemoglobin 4.2 mg/mL
Lotrimin Ultra cream 1.25% w/v
Monistat 7 cream 1.25% w/v
Nitrites 0.075 mg/mL
Plasma-lyte A 10% w/v
Vaginal contraceptive gel 1.25% w/v
Vagisil cream 1.25% w/v
Vagistat 1 cream 1.25% w/v

CROSS-REACTIVITY

Antibodies used in the MycoMEIA® Aspergillus assay detect galactofuranose-containing antigens, typically produced in high quantities by Aspergillus spp., other Ascomycetes fungi, and select other microorganisms, but not by mammals. Cross-reactivity was tested using healthy urine containing bacterial and fungal organisms, listed below. Testing was performed in duplicate (n=2). The MycoMEIA® assay gave a false positive result with Fusarium (8.80 x 10e5 CFU/mL), an Ascomycetes fungus. No microbial interference was observed in the presence of the organisms and the assay analyte.

Results from testing Contrived Samples Spiked with Microorganisms:

No cross-reactivity was observed in samples with the following:

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Acinetobacter baumanii 4.00 x 10e5 CFU/mL
Bacteroides fragilis 1.01 x 10e6 CFU/mL
Candida albicans 1.73 x 10e6 CFU/mL
Candida glabrata 1.91 x 10e6 CFU/mL
Candida parapsilosis 3.06 x 10e6 CFU/mL
Candida tropicalis 1.57 x 10e6 CFU/mL
Chlamydia trachomatis 7.00 x 10e5 CFU/mL
Citrobacter freundii 6.7 x 10e5 CFU/mL
Clostridia 5.28 x 10e5 CFU/mL
Corynebacterium amycolatum 2.80 x 10e5 CFU/mL
Cryptococcus neoformans 1.25 x 10e6 CFU/mL
Enterobacter cloacae 1.65 x 10e6 CFU/mL
Enterococcus faecalis 1.4 x 10e6 CFU/mL
Escherichia coli 1.15 x 10e6 CFU/mL
Geotrichum 2.80 x 10e5 CFU/mL
Klebsiella pneumoniae 6.9 x 10e5 CFU/mL
Lactobacillus crispatus 2.80 x 10e5 CFU/mL
Neisseria gonorrhoeae 3.90 x 10e5 CFU/mL
Parainfluenza virus 11.00 x 10e6 Virus/mL
Parainfluenza virus 25.00 x 10e5 Virus/mL
Parainfluenza virus 31.00 x 10e6 Virus/mL
Peptostreptococci 6.84 x 10e5 CFU/mL
Prevotella bivia 1.84 x 10e6 CFU/mL
Proteus mirabilis 1.35 x 10e6 CFU/mL
Pseudomonas aeruginosa 1.1 x 10e6 CFU/mL
Rhinovirus 2 2.00 x 10e6 Virus/mL

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Rhinovirus 83 4.50 x 10e5 Virus/mL
Serratia marcescens 1.1 x 10e6 CFU/mL
Staphylococcus aureus 1.37 x10e6 CFU/mL
Staphylococcus epidermidis 5.8 x 10 5 CFU/mL
Staphylococcus saprophyticus subsp. saprophyticus 2.60 x10 5 CFU/mL
Stenotrophomonas maltophilia 2 x 10 6 CFU/mL
Streptococcus agalactiae 1.37 x 10 6 CFU/mL

Cross-reactivity was also tested using urine samples obtained from people with different medical conditions, including fungal, viral, and bacterial infections. Cross-reactivity was detected from samples collected from people with histoplasmosis (n=1), blastomycosis (n=1), candidemia (n=1), streptococcus (n=2) and rhinovirus/parainfluenzavirus (n=1). One sample that showed a positive MycoMEIA test result (index ≥ 0.6) was identified as possible invasive aspergillosis.

Results from testing Clinical Samples:

UTI Streptococcus spp. Average IDX = 3.9
UTI Enterococcus spp., Enterobacter lung / blood Average IDX =0.5
UTI Streptococcus spp. Average IDX = 0.3
UTI Staphylococcus spp. Average IDX = 0.4
Klebsiella lung/blood Average IDX = 0.3
E. coli lung / blood Average IDX = 0.3
Possible IA + UTI Enterococcus, Enterococcus lung / blood Average IDX = 0.6
Enterobacter lung / blood, Streptococcus lung / blood Average IDX = 0.2
Streptococcus lung / blood Average IDX = 0.3
UTI Mixed gram positive Average IDX = 0.2
UTI Staphylococcus spp. Average IDX = 0.5
Acinetobacter junii lung / blood, URI – Rhinovirus, Parainfluenzavirus Average IDX = 0.3

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URI – Rhinovirus, Parainfluenzavirus Average IDX = 0.5
Streptococcus lung / blood Average IDX = 0.4
MSSA, Staphylococcus lung / blood, URI – Influenza Average IDX = 0.3
Staphylococcus lung / blood Average IDX = 0.3
Streptococcus lung / blood Average IDX = 0.6
P. aeruginosa lung / blood Average IDX = 0.3
MRSA, Staphylococcus lung / blood Average IDX = 0.3
Candida krusei Average IDX = 0.2
Candida krusei Average IDX = 0.2
MRSA, Staphylococcus lung / blood Average IDX = 0.2
URI – Rhinovirus, Parainfluenzavirus Average IDX = 0.2
E. coli lung / blood Average IDX = 0.1
Other IFI lung / blood Average IDX = 0.2
MRSA, Staphylococcus lung / blood Average IDX = 0.3
MRSA, Staphylococcus lung / blood Average IDX = 0.4
Histoplasmosis Average IDX = 0.2
Candida albicans (Candidiasis) Average IDX = 0.2
Pneumocystis jirovecii pneumonia Average IDX = 0.3
Other IFI lung / blood (cryptococcosis) Average IDX = 0.5
Histoplasmosis Average IDX = 0.3
Histoplasmosis Average IDX = 0.3
Histoplasmosis Average IDX = 0.3
Histoplasmosis Average IDX = 0.4
Histoplasmosis Average IDX = 18.6
Pneumocystis jirovecii pneumonia Average IDX = 0.3
Candidiasis Average IDX = 0.2

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Blastomyces lung / blood Average IDX = 0.6
Staphylococcus lung / blood Average IDX = 0.3
P. aeruginosa lung / blood Average IDX = 0.2
Pneumocystis jirovecii pneumonia Average IDX = 0.3
Candidiasis Average IDX = 0.6
MRSA, Stenotrophomonas lung / blood Average IDX = 0.4
P. aeruginosa lung / blood Average IDX = 0.3
Fusariosis lung / blood Average IDX = 0.2
P. aeruginosa lung / blood Average IDX = 0.3

Additional cross-reactivity testing was performed using clinical samples associated with episodes of mixed infections and non-Aspergillus infections. All subjects were at risk for IA by virtue of having current or recent episodes of neutropenia (e.g., due to cytotoxic therapy), receipt of allogeneic BMT, receipt of solid organ transplant (kidney, heart, liver), receipt of other biologic immunosuppressants for autoimmune disease, inherited severe immunodeficiency or another underlying disease that portends high-risk for IA (e.g., HIV/AIDS, severe malnutrition). Subjects in the clinical study were inpatients and had at least 2 weeks of clinical follow-up to determine clinical diagnosis, with at least 4 urine samples obtained during the clinical evaluation period or at least 1 sample obtained within 1 week of diagnosis.

Of 21 subjects with documented bacterial pneumonia (18 with concurrent bacteremia) no urine samples tested positive with MycoMEIA. Cross-reactivity was detected from samples collected from people with mixed bacterial infection (n=1), mixed gastrointestinal complications (n=2), histoplasmosis (n=2), blastomycosis (n=1), candidemia (n=1), and disseminated fusariosis (n=1). Positivity was more frequent in people who had ambiguous diagnoses. These included people considered to have 'possible IA' with specific radiographic findings, but with bacterial cultures positive in cultures other than BAL (e.g., sputum or blood).

These findings show that cross-reactive MycoMEIA tests can occur with infections caused by other Ascomycetes fungi.

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CLINICAL PERFORMANCE

The clinical performance of MycoMEIA Aspergillus assay (MycoMEIA) was evaluated using clinical samples obtained from existing archives, and a prospective study, powered to evaluate the specificity of fresh samples.

In the archived, retrospective study, a total of 475 samples collected from 290 subjects were analyzed. 226 samples were tested from 50 subjects with invasive aspergillosis (IA) that was proven (n = 3) or probable (n = 47) per 2020 EORTC/MSG diagnostic criteria.

A prospective study was performed to analyze the specificity of the assay in fresh samples. Testing of the clinical samples was performed within 6 hours of collection. The study was performed to include all consenting inpatients who had blood for GM EIA testing sent to the lab, regardless of age, underlying disease, or reason for clinical suspicion. Lung transplant recipients and pediatric patients are not indicated for use with the MycoMEIA and were excluded from analyses. Because the definition of 'no IA' vs. 'possible IA' is contingent on results of chest CT, subjects who did not have CT performed within 2 weeks of urine sampling were also excluded from analyses. Clinical diagnoses were adjudicated by a reviewer who was blinded to MycoMEIA test results. Diagnoses included proven IA, probable IA, and possible IA, as per the 2020 EORTC/MSG definitions.

A total of 210 subjects were consented and provided 254 urine samples for testing during 213 different suspected infection episodes. Of these, 34 subjects had lung transplants and were excluded from IA analysis. 2 pediatric subjects were also excluded. Of 177 remaining episodes of suspected IA, 8 more were excluded. One sample was noted by visual appearance in the laboratory to be likely contaminated prior to testing. Three subjects did not have CT scans performed during the 2-week interval around sample collection. Four subjects were discontinued because samples were collected after receipt of > 3 days of mold-active antifungal therapy. Final analysis was performed with 166 subjects and 169 infectious episodes.

"Possible" IA (n=106) was the most common clinical adjudication in the prospective study and were excluded from the analysis. Thirty-eight (38) subjects were adjudicated as having no invasive fungal infection. Twenty-six (26) subjects were adjudicated as having mixed or other (non-IA) infections.

Across the studies, MycoMEIA Index values ranged from 0.1 to 45.0 in the IA population. Performance of the assay was estimated as a per-subject sensitivity of 92.4% (95% CI 82.1-97.0%) and per-sample sensitivity of 58.2% (95% 54.3 – 61.9%).

The mean MycoMEIA Index value for the population with no IA was 0.5. The specificity in the prospective study was 86.1% (95% CI 75.7-92.5%) using the Positive cut-off (≥ 0.6).

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	MycoMEIA Positive	MycoMEIA Negative	Sensitivity (95% CI)
Positive Subjects
Cases, proven and probable IA (N=53)	49	4	92.4% (82.1-97.0)
Cases, proven IA, archived (n=3)	3	0	100% (43.8-100)
Cases, probable IA, archived (n=44)	41	3	93.2% (81.8-97.7)
Cases, probable IA, prospective (n=6)	5	1	83.3% (43.6-97.0)
Positive Samples
Samples, proven and probable IA (N=636)	370	266	58.2% (54.3-61.9)
Samples, proven IA, archived (n = 72)	46	26	63.9% (52.3-74.0)
Samples, probable IA, archived (n=551)	318	233	57.7% (53.5-61.8)
Samples, probable IA, prospective (n=13)	6	7	46.1% (23.2-70.9)

	MycoMEIA Positive	MycoMEIA Negative	Specificity (95% CI)
Negative Samples (Prospective Study)
Prospective (N=65)	9	56	86.1% (75.7-92.5)
No infection (n=39)	4	34	89.5% (75.9-95.8)
Mixed or other infections (n=26)	5	21	80.8% (62.1-91.5)

HIGH-DOSE HOOK EFFECT

No high-dose hook effect was observed with Aspergillus antigen ethanol precipitation (EP) concentrations up to 1,000 ng/mL.

Regulation Number and Section

§ 866.3040

Aspergillus spp. serological reagents.(a)
Identification. Aspergillus spp. serological reagents are devices that consist of antigens and antisera used in various serological tests to identify antibodies toAspergillus spp. in serum. The identification aids in the diagnosis of aspergillosis caused by fungi belonging to the genusAspergillus. Aspergillosis is a disease marked by inflammatory granulomatous (tumor-like) lessions in the skin, ear, eyeball cavity, nasal sinuses, lungs, and occasionally the bones.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 866.9.