(426 days)
Not Found
No
The device description details a standard immunoassay with optical density measurement and curve fitting, which does not involve AI/ML. The document explicitly states "Mentions AI, DNN, or ML: Not Found".
No
Explanation: This device is an in-vitro diagnostic (IVD) test used to aid in the diagnosis of histoplasmosis by detecting antigens. It does not provide therapy or treatment for the condition.
Yes
The "Intended Use / Indications for Use" section explicitly states that the device "can be used as an aid in the diagnosis of histoplasmosis." This phrasing clearly indicates its role in the diagnostic process.
No
The device is a laboratory assay kit that includes physical reagents and microwell plates, not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it's for the "detection of Histoplasma antigens in urine samples" and is used "as an aid in the diagnosis of histoplasmosis." This clearly indicates it's a test performed on a sample taken from the human body to provide information for diagnostic purposes.
- Device Description: The description details a laboratory-based assay (immunoenzymatic sandwich microplate assay) that uses reagents to detect a specific analyte (Histoplasma antigens) in a biological sample (urine). This is characteristic of an in vitro diagnostic test.
- Performance Studies: The document describes clinical performance evaluations using human urine specimens and provides metrics like sensitivity and specificity, which are standard for evaluating the performance of IVD devices.
- Predicate Device: The mention of a predicate device (Bio-Rad's Platelia™ Aspergillus EIA) is common in regulatory submissions for IVDs, indicating a comparison to an already cleared IVD.
All these elements align with the definition and characteristics of an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The ALPHA Histoplasma Antigen ElA is an immunoenzymatic sandwich microplate assay for the detection of Histoplasma antigens in urine samples.
The ALPHA Histoplasma Antigen 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 histoplasmosis.
Product codes
MIZ
Device Description
The ALPHA Histoplasma Antigen EIA is an immunoenzymatic sandwich microplate assay which detects Histoplasma antigens in urine. Rabbit anti-Histoplasma IgG antibodies bound to microwell plates are used as capture antibodies and biotinylated rabbit anti-Histoplasma IgG antibodies are used as detect antibodies.
The positive control is made of buffer spiked with Histoplasma capsulatum antigens and the negative control is buffer only. Standards are made of Histoplasma capsulatum antigens from culture filtrate. The kit contains the 100 Standard, 30 Standard, and 2 Standard that are used to generate a sigmoid calibration curve using a four-parameter fit of the blanked absorbance values versus the assigned EIA Values. R-squared values must be greater than or equal to 0.990.
Urine samples are run untreated and undiluted. The samples are added to the microwells coated with the capture antibody and incubated. If the patient specimen contains Histoplasma antigens that are recognized by the capture antibody, those antigens will become bound to the microwell. The wells are washed to remove unbound patient material and biotinylated detection antibody is added to the wells. If Histoplasma antigens are bound to the microwell by the capture antibody, then the detect antibody will also become bound to the microwell. The wells are then washed to remove any unbound detect antibody. For detection, biotinstreptavidin chemistry reagents including 3,3′,5′,5′ tetramethybenzadine (TMB) and stop solution are used. Streptavidin conjugated to horseradish peroxidase (HRP) is added to the microwells. In the presence of the biotinylated detect antibody, streptavidin-HRP will become bound to the plate. The plate is then washed to remove any unbound streptavidin-HRP, and TMB substrate solution is added to the microwells. A blue color develops in the presence of the HRP enzyme. The reaction is stopped by the addition of a stop solution. The optical density (absorbance) is determined with a microplate reader at 450nm or 450nm or 450nm alone. ElA Units for specimens are calculated using a four-parameter curve-fit generated with the 4 standards supplied in the kit.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Not Found
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies
The clinical performance of the ALPHA Histoplasma Antigen EIA was evaluated using a total of 278 culture- or histopathology-confirmed urine specimens.
Comparison to Other Histoplasma Antigen EIA: Supplemental comparison studies were performed using a non-FDA-cleared Histoplasma capsulatum quantitative antigen EIA.
Key Metrics
Comparison to Culture/Histopathology:
Sensitivity: 80.9% (95% CI: 67.5-89.6%)
Specificity: 98.7% (95% CI: 96.3-99.6%)
Comparison to Other Histoplasma Antigen EIA:
Equivocals Assumed Positive:
% Agree Positive: 71.4% (95% CI: 57.6-82.2%)
% Agree Negative: 94.0% (95% CI: 83.8-97.9%)
Equivocals Assumed Negative:
% Agree Positive: 92.1% (95% CI: 79.2-97.3%)
% Agree Negative: 95.1% (95% CI: 86.5-98.3%)
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 866.3320
Histoplasma capsulatum serological reagents.(a)
Identification. Histoplasma capsulatum serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies toHistoplasma capsulatum in serum. Additionally, some of these reagents consist ofHistoplasma capsulatum antisera conjugated with a fluorescent dye (immunofluorescent reagents) used to identifyHistoplasma capsulatum from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of histoplasmosis caused by this fungus belonging to the genusHistoplasma and provides epidemiological information on the diseases caused by this fungus. Histoplasmosis usually is a mild and often asymptomatic respiratory infection, but in a small number of infected individuals the lesions may spread to practically all tissues and organs.(b)
Classification. Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 866.9.
0
JUL 19 2011
510(k) Premarket Notification ALPHA Histoplasma Antigen EIA
510(k) Summary ALPHA Histoplasma Antigen EIA
This 510(k) summary is submitted in accordance with 21 CFR §807.92
Assigned 510(k) No .: K101407
Owner: Immuno-Mycologics, Inc. 2700 Technology Place Norman, OK 73071 Tel: 405-360-4669 Fax: 405-364-1058 Contact: Dr. Sean K. Bauman, President & CEO July 7, 2011 Prepared:
Trade Name: ALPHA Histoplasma Antigen EIA
Histoplasma Antigen EIA Common Name:
Classification Name: None
Regulation: 866.3320
Bio-Rad's Platelia™ Aspergillus EIA, K060641 Predicate Device:
Intended Use: The ALPHA Histoplasma Antigen ElA is an immunoenzymatic sandwich microplate assay for the detection of Histoplasma antigens in urine samples.
The ALPHA Histoplasma Antigen 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 histoplasmosis.
Device Description:
The ALPHA Histoplasma Antigen EIA is an immunoenzymatic sandwich microplate assay which detects Histoplasma antigens in urine. Rabbit anti-Histoplasma IgG antibodies bound to microwell plates are used as capture antibodies and biotinylated rabbit anti-Histoplasma IgG antibodies are used as detect antibodies.
The positive control is made of buffer spiked with Histoplasma capsulatum antigens and the negative control is buffer only. Standards are made of Histoplasma capsulatum antigens from culture filtrate. The kit contains the 100 Standard, 30 Standard, and 2 Standard
Rev. 07/11/2011
Page 5-1
1
that are used to generate a sigmoid calibration curve using a four-parameter fit of the blanked absorbance values versus the assigned EIA Values. R-squared values must be greater than or equal to 0.990.
Urine samples are run untreated and undiluted. The samples are added to the microwells coated with the capture antibody and incubated. If the patient specimen contains Histoplasma antigens that are recognized by the capture antibody, those antigens will become bound to the microwell. The wells are washed to remove unbound patient material and biotinylated detection antibody is added to the wells. If Histoplasma antigens are bound to the microwell by the capture antibody, then the detect antibody will also become bound to the microwell. The wells are then washed to remove any unbound detect antibody. For detection, biotinstreptavidin chemistry reagents including 3,3′,5′,5′ tetramethybenzadine (TMB) and stop solution are used. Streptavidin conjugated to horseradish peroxidase (HRP) is added to the microwells. In the presence of the biotinylated detect antibody, streptavidin-HRP will become bound to the plate. The plate is then washed to remove any unbound streptavidin-HRP, and TMB substrate solution is added to the microwells. A blue color develops in the presence of the HRP enzyme. The reaction is stopped by the addition of a stop solution. The optical density (absorbance) is determined with a microplate reader at 450nm or 450nm or 450nm alone. ElA Units for specimens are calculated using a four-parameter curve-fit generated with the 4 standards supplied in the kit.
Comparison with Predicate:
A comparison of the similarities and differences between the ALPHA Histoplasma Antigen ElA and the predicate device is presented in the table below (Table 1).
Feature | ALPHA Histoplasma Ag EIA Device | Platelia Aspergillus EIA K060641 |
---|---|---|
Intended Use | Antigen detection | Antigen detection |
Indication For Use | Aid in the diagnosis | Aid in the diagnosis |
Device Description | ||
Assay Principle | EIA | EIA |
Assay components | 96-well microplate coated antibody, wash buffer, positive control, negative control, enzyme conjugate, TMB substrate, stop solution | 96-well microplate coated antibody, wash buffer, positive control, negative control, enzyme conjugate, TMB substrate, stop solution |
Detection Chemistry | HRP + TMB | HRP + TMB |
Controls/Standard | Antigen | Antigen |
Instruments | none | none |
Microplate | 96-well microplate coated with antibody | 96-well microplate coated with antibody |
Performance Characteristics | ||
Precision | Acceptable % CVs | Acceptable % CVs |
Table 1. Similarities and Differences between ALPHA Histoplasma EIA and Platelia Aspergillus ElA | |
---|---|
-------------------------------------------------------------------------------------------------- | -- |
2
Linearity | Not applicable | Not applicable |
---|---|---|
Specificity | Cross-reacts with other fungal organisms | Cross-reacts with other fungal organisms |
Feature | ALPHA Histoplasma Ag EIA Device | Platelia Aspergillus EIA K060641 |
---|---|---|
Intended Use | Detection of Histoplasma antigen in urine samples | Detection of Aspergillus galactomannan antigen in |
adult and pediatric serum samples | ||
Indication For Use | Aid in the diagnosis of histoplasmosis. | Aid in the diagnosis of invasive aspergillosis. |
Sample Matrix | Urine | Serum |
Controls | Histoplasma antigens | Aspergillus galactomannan |
Output | "EIA Units" as determined from standard curve | "Index" as determined by OD of sample divided by |
Cut-Off Control OD | ||
Detection | ||
Antibody | Biotinylated anti-Histoplasma polyclonal antibody | HRP-linked anti-Aspergillus monoclonal antibody |
Microplate | 96-well microplate coated with anti-Histoplasma | |
polyclonal antibody | 96-well microplate coated with anti-Aspergillus | |
monoclonal antibody | ||
Assay Time | ~3 hours | ~2 hours |
Performance | ||
Characteristics | ||
Shelf life | 1 year | Each kit component different |
Analytical Performance Summary
- A. Urine Precision Studies
Three sites, a reference laboratory (RL) (Western US), a clinical laboratory (CL) (Upper Mid-Western US), and IMMY (Central US), were used to assess the assay's reproducibility. The panel consisted of urine samples at five levels: negative (روم), cut-off (دول), low positive (C35), and moderately positive. At the reference laboratory and at IMMY, each sample was tested in triplicate over the course of 5 days, using multiple operators. At the clinical laboratory, each sample was tested in triplicate over the course of 3 days, using a single operator. Throughout the study, reagent lots and instrument calibrations were held constant. No runs were removed from analysis due to failed runs.
Variance was estimated by calculating the mean value of each sample, the standard deviation and percent CV. The data was analyzed separately to evaluate any inter-assay, intra-assay, and inter-site variation. Overall, no major source of variability was identified. Intra-run, Inter-Run, and Inter-site percent CVs are within acceptable limits (≤ 20%), with the exception of the negative urine and low negative urine samples, which is expected when testing beyond the limit of detection. A summary of the data is reported in the Tables 2-4 below.
3
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: :
Table 2. Intra-run Reproducibility Analysis | |||||||||
---|---|---|---|---|---|---|---|---|---|
Intra-run Analysis Blanked OD Values | |||||||||
Neg. | |||||||||
Urine | C5 | C50 | C95 | Moderate | |||||
Urine | Positive | ||||||||
Control | |||||||||
Clinical Laboratory | Day 1 | Operator 1 | Ave | 0.010 | 0.064 | 0.078 | |||
SD | 0.003 | 0.001 | 0.002 | 0.003 | 0.032 | 0.008 | |||
%CV | 26.5% | 1.8% | 2.0% | 3.3% | 3.9% | 3.4% | |||
Day 2 | Operator 1 | Ave | 0.006 | 0.071 | 0.088 | 0.108 | 0.900 | 0.246 | |
SD | 0.003 | 0.002 | 0.003 | 0.003 | 0.032 | 0.004 | |||
%CV | 50.0% | 2.9% | 3.4% | 2.8% | 3.6% | 1.5% | |||
Day 3 | Operator 1 | Ave | 0.007 | 0.062 | 0.074 | 0.086 | 0.783 | 0.211 | |
SD | 0.002 | 0.002 | 0.004 | 0.007 | 0.041 | 0.009 | |||
%CV | 31.2% | 3.4% | 5.1% | 8.1% | 5.3% | 4.3% | |||
Reference Laboratory | Day 1 | Operator 1 | Ave | 0.006 | 0.035 | 0.054 | 0.057 | 0.523 | 0.136 |
SD | 0.003 | 0.002 | 0.001 | 0.007 | 0.011 | 0.008 | |||
%CV | 39.7% | 4.3% | 1.9% | 11.3% | 2.1% | 5.6% | |||
Day 2 | Operator 2 | Ave | -0.003 | 0.039 | 0.065 | 0.061 | 0.612 | 0.172 | |
SD | 0.006 | 0.002 | 0.001 | 0.002 | 0.024 | 0.008 | |||
%CV | 183% | 4.0% | 0.9% | 2.5% | 3.9% | 4.9% | |||
Day 3 | Operator 3 | Ave | 0.011 | 0.048 | 0.072 | 0.064 | 0.642 | 0.182 | |
SD | 0.001 | 0.003 | 0.004 | 0.009 | 0.078 | 0.007 | |||
%CV | 9.1% | 5.2% | 4.9% | 13.5% | 12.2% | 3.9% | |||
Day 4 | Operator 4 | Ave | -0.001 | 0.045 | 0.071 | 0.077 | 0.654 | 0.195 | |
SD | 0.010 | 0.005 | 0.003 | 0.007 | 0.018 | 0.005 | |||
%CV | 766% | 11.8% | 4.3% | 9.1% | 2.7% | 2.4% | |||
Day 5 | Operator 4 | Ave | 0.007 | 0.042 | 0.061 | 0.069 | 0.620 | 0.142 | |
SD | 0.001 | 0.002 | 0.015 | 0.002 | 0.031 | 0.011 | |||
%CV | 17.3% | 3.7% | 24.5% | 2.2% | 5.0% | 8.0% | |||
IMMY | Day-1 | Operator 1 | Ave | 0.012 | 0.049 | 0.070 | 0.081 | 0.686 | 0.204 |
SD | 0.003 | 0.010 | 0.014 | 0.011 | 0.039 | 0.027 | |||
%CV | 24.2% | 20.6% | 19.4% | 13.6% | 5.7% | 13.0% | |||
Day 2 | Operator,1 | Ave | 0.004 | 0.039 | 0.068 | 0.066 | 0.623 | 0.226 | |
SD | 0.002 | 0.002 | 0.010 | 0.005 | 0.043 | 0.028 | |||
%CV | 57.1% | 4.5% | 15.1% | 7.6% | 6.8% | 12.5% |
Tahle 2 Intra-run Reproducibility Analysis
ﺮ
・一
4
| Day 3
Operator 1 | Ave | 0.003 | 0.045 | 0.067 | 0.076 | 0.723 | 0.216 |
---|---|---|---|---|---|---|---|
SD | 0.001 | 0.006 | 0.009 | 0.004 | 0.051 | 0.004 | |
%CV | 36.4% | 13.6% | 13.3% | 5.3% | 7.0% | 1.6% | |
Day 4 | |||||||
Operator 2 | Ave | 0.008 | 0.043 | 0.064 | 0.068 | 0.653 | 0.202 |
SD | 0.003 | 0.006 | 0.009 | 0.006 | 0.025 | 0.017 | |
%CV | 38.6% | 15.1% | 14.7% | 8.1% | 3.8% | 8.4% | |
Day 5 | |||||||
Operator 2 | Ave | 0.005 | 0.042 | 0.072 | 0.080 | 0.663 | 0.228 |
SD | 0.002 | 0.005 | 0.008 | 0.008 | 0.034 | 0.006 | |
%CV | 31.6% | 12.2% | 11.3% | 10.3% | 5.1% | 2.7% |
Table 3. Inter-Run Reproducibility Analysis
Inter-Run Analysis - Blanked OD Values | |||||||
---|---|---|---|---|---|---|---|
Neg. | |||||||
Urine | C5 | C50 | C95 | Moderate | |||
Urine | Positive | ||||||
Control | |||||||
Ave | 0.008 | 0.066 | 0.080 | 0.095 | 0.831 | 0.225 | |
Std. Dev. | 0.003 | 0.005 | 0.007 | 0.011 | 0.061 | 0.017 | |
% CV | 38.7% | 7.0% | 8.6% | 11.0% | 7.4% | 7.6% | |
Reference | |||||||
Laboratory | Ave | 0.004 | 0.042 | 0.064 | 0.066 | 0.610 | 0.165 |
Std. Dev. | 0.007 | 0.005 | 0.009 | 0.009 | 0.059 | 0.025 | |
% CV | 187.0% | 12.7% | 13.9% | 13.1% | 9.6% | 14.9% | |
IMMY | Ave | 0.006 | 0.043 | 0.068 | 0.074 | 0.670 | 0.215 |
Std. Dev. | 0.004 | 0.006 | 0.009 | 0.009 | 0.048 | 0.020 | |
% CV | 64.2% | 14.9% | 13.2% | 11.7% | 7.2% | 9.1% |
Table 4. Inter-Site Reproducibility Analysis
Inter-Site Analysis - Blanked OD Values | ||||||
---|---|---|---|---|---|---|
Neg. | ||||||
Urine | C5 | C50 | C95 | Moderate | ||
Urine | Positive | |||||
Control | ||||||
Ave | 0.006 | 0.048 | 0.069 | 0.076 | 0.684 | 0.198 |
Std. Dev. | 0.005 | 0.011 | 0.010 | 0.014 | 0.101 | 0.034 |
% CV | 95.9% | 23.7% | 14.8% | 19.1% | 14.8% | 17.0% |
- B. Analytical Sensitivity (Lower limits of the assay)
Analytical sensitivity was determined (according to CLSI EP17-A) by determining the assay's Limit of the Blank and Limit of Detection. Wash buffer was spiked with Histoplasma antigens at a concentration range of 1 – 4 EIA Units. To determine the LoB, 82 replicates of wash buffer were assayed. To determine the LoD, 30 replicates of the spiked wash buffer were assayed. The Limit of the Blank is 0.009 OD and the Limit of Detection is 2 EIA Units (0.044 Blanked OD).
5
C. Analytical Specificity (Cross-Reactivity)
Urine specimens that tested negative for Histoplasma antigen were spiked with antigen from Blastomyces dermatitidis, Coccidioides immitis, Aspergillus sp., Paracoccidioides brasilliensis, Candida albicans, and Cryptococcus neoformans, individually at 1ug/ml. The ALPHA Histoplasma Antigen EIA is found to be cross-reactive with Blastomyces dermatitidis, Coccidioides immitis, and Paracoccidioides brasilliensis. The assay is not cross-reactive with Candida albicans, Cryptococcus neoformans, or Aspergillus sp. in urine. Furthermore, Aspergillus, Candida, Paracoccidioides, and Penicillium culture-positive urine specimens were tested in the ALPHA Histoplasma Antigen EIA. The results are summarized in Table 5.
Table 5. Specificity Using Culture-Confirmed Urine Specimens
Percent Positive | |
---|---|
Aspergillus | 0% (0/20) |
Candida spp. | 0% (0/12) |
Paracoccidioides | 9.1% (1/11) |
Penicillium | 0% (0/2) |
-
D. Analytical Interference
To evaluate substances that could potentially interfere with the ALPHA Histoplasma Antigen EIA, urine specimens containing various substances were obtained from a national reference laboratory. Substances included protein, blood, epithelial cells, ketones, mucus, casts, glucose, and bilirubin. Each specimen was spiked with Histoplasma antigen and tested. None of these substances were found to interfere with the ALPHA Histoplasma Antigen EIA. Vaginal cream urines and foods which produce color in urine were not tested. Additionally, drugs, such as itraconazole, amphotericin B, acetaminophen, acetylsalicylic acid, ascorbic acid, and caffeine were not tested for interference. -
E. Carry-over
To examine potential well-to-well carry-over, a positive sample was used in series alternating with a negative sample in a checkerboard pattern. Samples were found not to carry-over when methods described in the package insert were followed. -
F. Prozone Effect
To detect the prozone effect in the ALPHA Histoplasma Antigen EIA, negative urine was spiked with Histoplasma antigen to 2000, 3000, and 4000 EIA Units and tested. All samples remained positive, therefore, prozoning is not seen in the ALPHA Histoplasma Antigen EIA. -
G. Linearity
N/A
6
H. Assay cut-off
-------. ..
The assay's cut-off of 2.0 EIA Units was established by Limit of Detection (Section B) experiments and confirmed with the determination of the C50. Receiver Operator Curve (ROC) analysis of culture-proven specimens indicated a 1.3 ElA Unit cutoff. However, this is below the Limit of Detection (2.0 EIA units), and therefore an inappropriate cutoff.
Negative: ✓ |
|-----------------------------|-----------------------------------------------------------------------------------------|
| (Part 21 CFR 801 Subpart D) | |
Over-The-Counter Use AND/OR (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Freddie K. Poole
Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety
510(k): K101407