The ALPHA Histoplasma Antigen EIA 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 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, biotin-streptavidin 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. EIA Units for specimens are calculated using a four-parameter curve-fit generated with the 4 standards supplied in the kit.

AI/ML Overview

Here's an analysis of the acceptance criteria and the study that proves the device meets them, based on the provided text:

ALPHA Histoplasma Antigen EIA Performance Study Analysis

The ALPHA Histoplasma Antigen EIA is an immunoenzymatic sandwich microplate assay designed for the detection of Histoplasma antigens in urine samples, serving as an aid in the diagnosis of histoplasmosis when combined with other diagnostic procedures.

1. Acceptance Criteria and Reported Device Performance

The direct acceptance criteria for this device are not explicitly stated in a consolidated manner as pass/fail thresholds in the provided document. However, the studies conducted demonstrate acceptable performance characteristics. Based on the "Comparison to Culture/Histopathology" section, two key performance metrics were evaluated: Sensitivity and Specificity.

Acceptance Criteria (Implied)	Reported Device Performance
Sensitivity (acceptable)	80.9% (95% CI: 67.5-89.6%)
Specificity (acceptable)	98.7% (95% CI: 96.3-99.6%)

Note: The document implies "acceptable" performance for these metrics rather than specifying strict numerical thresholds for acceptance prior to the study.

2. Sample Size and Data Provenance

Test Set Sample Size: A total of 278 urine specimens were used for the clinical performance evaluation (comparison to culture/histopathology).
Data Provenance: The document does not explicitly state the country of origin for the clinical samples. It does mention that "urine specimens containing various substances were obtained from a national reference laboratory" for analytical interference studies, suggesting some US-based data. However, for the main clinical performance, the geographic origin is not specified. The data is retrospective, as specimens were "culture- or histopathology-confirmed" prior to testing with the device.

3. Number of Experts and Qualifications for Ground Truth

The document does not explicitly state the number of experts used to establish the ground truth or their specific qualifications (e.g., "radiologist with 10 years of experience"). However, the ground truth was established by "culture- or histopathology-confirmed" results. This implies that the diagnoses were made by medical professionals, likely pathologists and/or microbiologists, following standard diagnostic procedures.

4. Adjudication Method for the Test Set

The document does not describe a specific adjudication method (e.g., 2+1, 3+1) for establishing the ground truth of the clinical test set. The ground truth was based on "culture- or histopathology-confirmed" results, which are considered definitive diagnostic methods. This suggests that individual confirmation by these methods was sufficient, and a separate adjudications process was not deemed necessary for the diagnostic truth.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not performed. The study evaluates the performance of the device against established diagnostic methods (culture/histopathology) and against another, non-FDA-cleared Histoplasma antigen EIA, but it does not assess human reader improvement with or without AI assistance.

6. Standalone Performance Study

Yes, a standalone performance study was done. The "Comparison to Culture/Histopathology" section directly assesses the algorithm's (device's) ability to detect Histoplasma antigens in urine samples against established clinical diagnostic methods, without human interpretation as part of the primary outcome.

7. Type of Ground Truth Used

The type of ground truth used was expert consensus, pathology, and microbiological culture results. Specifically, the clinical performance was evaluated against "culture- or histopathology-confirmed urine specimens."

8. Sample Size for the Training Set

The document does not provide information regarding the sample size used for the training set. This is a 510(k) submission, typically focusing on validation rather than development details.

9. How Ground Truth for the Training Set Was Established

The document does not provide information on how the ground truth for any potential training set was established. Given this is a 510(k) for an immunoassay, the device itself is a diagnostic test, and the development process likely involved internal validation and optimization, but specific training set details are not included in this regulatory summary.

Summary

{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 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 inadult 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 byCut-Off Control OD
DetectionAntibody	Biotinylated anti-Histoplasma polyclonal antibody	HRP-linked anti-Aspergillus monoclonal antibody
Microplate	96-well microplate coated with anti-Histoplasmapolyclonal antibody	96-well microplate coated with anti-Aspergillusmonoclonal antibody
Assay Time	~3 hours	~2 hours
PerformanceCharacteristics
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}------------------------------------------------

、

: :

Table 2. Intra-run Reproducibility Analysis
Intra-run Analysis Blanked OD Values
			Neg.Urine	C5	C50	C95	ModerateUrine	PositiveControl
			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 3Operator 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 4Operator 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 5Operator 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	ModerateUrine	PositiveControl
	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%
ReferenceLaboratory	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	ModerateUrine	PositiveControl
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: < 2.0 EIA Units Positive: ≥2.0

Single Versus Dual Wavelength Analysis 1.
Eight positive urine samples and the 4 standards were tested in duplicate and according to the package insert. The microwells were read at 450/630 nm and then at 450 nm only. The study confirmed that the blanked OD of a sample is not affected by the read method, as indicated by the low percent CVs across both read Furthermore, 450/630 vs. 450 alone can be used with equal accuracy, methods. and data from the two methods can be used interchangeably.
J. Reading the Test
To demonstrate that the test should be read within 15 minutes after the addition of the Stop Solution, all the standards were run in triplicate according to the package insert. The test was then read immediately after the addition of the Stop Solution, and then at 15, 30, 45, 60, 75, and 90 minutes. The signal began to decrease after 15 minutes and continued to decrease over the course of 90 minutes. While the decrease was not significant, it is still preferable to read the assay as soon after the addition of the Stop Solution as possible to avoid errors.
K. Specimen Acceptance Criteria
In specimen storage studies, reductions in EIA values after 2-week storage at 4°C and after multiple freeze thaws were observed. However, no specimen went from positive to negative (Figures 1 and 2). Since a reduction in EIA values was observed, it is possible that a fresh, very low-positive specimen (near 2.0 EIA units) could become negative if it is stored for several days.

Rev. 07/11/2011

{7}------------------------------------------------

510(k) Premarket Notification ALPHA Histoplasma Antigen EIA

Immuno-Mycologics, Inc Norman, OK

... .

Image /page/7/Figure/2 description: The image is a line graph titled "4° Storage". The x-axis is labeled "Day" and ranges from 0 to 15. The y-axis is labeled "EIA Units" and ranges from 0.0 to 25.0. There are four lines on the graph, labeled "Sample 1", "Sample 2", "Sample 3", and "Sample 4", each representing a different sample's EIA Units over time.

Figure 1. EIA Units Versus Days Stored at 4°C

Image /page/7/Figure/4 description: The image is a line graph titled "-20° Freeze/Thaw Storage". The x-axis is labeled "Day" and ranges from 0 to 15. The y-axis is labeled "EIA Units" and ranges from 0.0 to 30.0. There are four lines on the graph, labeled "Sample 1", "Sample 2", "Sample 3", and "Sample 4", which represent the change in EIA Units over time.

Figure 2. EIA Units Versus Days Stored at -20°C

L. Method Comparison

Comparison to Culture/Histopathology:

The clinical performance of the ALPHA Histoplasma Antigen EIA was evaluated using a total of 278 culture- or histopathology-confirmed urine specimens. The results are summarized in Tables 6 and 7.

Table 6. 2x2 Contingency Table Comparing Device to Culture/Histopathology

		Culture/Histopathology
		Positive	Negative
IMMY ALPHA	Positive	38	3
Histoplasma EIA	Negative	9	228

{8}------------------------------------------------

510(k) Premarket Notification ALPHA Histoplasma Antigen EIA

Table 7. Statistical Analysis: Device vs Culture/Histopathology

	PointEstimate	95% CI
Sensitivity	80.9%	67.5-89.6%
Specificity	98.7%	96.3-99.6%

Comparison to Other Histoplasma Antigen EIA

Supplemental comparison studies were performed using a non-FDA-cleared Histoplasma copsulatum quantitative antigen EIA. The other test has an equivocal range and therefore, analysis was performed by either considering the equivocals positive or negative. The resulting 2x2 contingency tables (Tables 8 and 10) and statistical analysis (Tables 9 and 11) are below.

Table 8. 2x2 Contingency Table Comparing Device to other EIA; Equivocals Assumed Positive

		Other Histo Antigen EIA
		Positive	Negative
IMMY ALPHA	Positive	35	3
Histoplasma EIA	Negative	14	47

Table 9. Statistical Analysis

	PointEstimate	95% CI
% Agree Positive	71.4%	57.6-82.2%
% Agree Negative	94.0%	83.8-97.9%

Table 10. 2x2 Contingency Table Comparing Device to other EIA; Equivocals Assumed Negative

		Other Histo Antigen EIA
		Positive	Negative
IMMY ALPHA	Positive	35	3
Histoplasma EIA	Negative	3	58

Table 11. Statistical Analysis

	PointEstimate	95% CI
% Agree Positive	92.1%	79.2-97.3%
% Agree Negative	95.1%	86.5-98.3%

Conclusion

ﺘﺮﺗﺒﻪ ﺍﻟﺘﺮﺗﻴﺔ ﺍﻟﺘﻲ ﺗﺘﺮﺗﺒﺎﺕ ﺍﻟﺘﻲ ﺗﺘﺮﺗﺒﺎﺕ ﺍﻟﻤﺘﺮﻭ ﺍﻟﺘﻲ ﺗﺮﺗﺒﺎﺕ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟﻤﺘﺮﻭ ﺍﻟ

The information submitted in this premarket notification is complete and supports a substantial equivalence decision.

{9}------------------------------------------------

DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/9/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with three wavy lines representing its wings and tail. The eagle is positioned within a circular border that contains the text "DEPARTMENT OF HEALTH & HUMAN SERVICES · USA" in a sans-serif font.

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

Immuno-Mycologics, Inc c/o Sean K. Bauman, Ph.D. President and CEO 2700 Technology Place JUL 1 9 2011 Norman, OK 73071 Re: K101407 Trade/Device Name: ALPHA Histoplasma Antigen ELA Regulation Number: 21CFR §866.3320 Regulation Name: Histoplasma capsulatum serological reagents. Regulatory Class: Class II Product Code: MIZ July 11, 2011 Dated: Received: July 12, 2011

Dear Dr. Bauman:

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 (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. 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.

If your device is classified (see above) into class II (Special Controls), it may be subject to such 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.

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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. This letter will allow you to begin marketing your device as described in your Section

{10}------------------------------------------------

Page 2 - Sean K. Bauman, Ph.D.

510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Vagant

Sally A. Hojvat, M.Sc. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

{11}------------------------------------------------

510(k) Premarket Notification ALPHA Histoplasma Antigen EIA

Indications for Use Statement

510(k) Number (if known): K101407

Device Name: ALPHA Histoplasma Antigen ElA

Indications for Use:

The ALPHA Histoplasma Antigen EIA is an immunoenzymatic sandwich microplate assay for the detection of Histoplasma antigens in urine samples.

The ALPHA Histoplasma Antigen ElA 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.

Prescription Use	✓
(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

Regulation Number and Section

§ 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.