Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device is an immunoenzymatic assay, which is a biochemical test, not a software-based technology that would typically incorporate AI/ML. The description focuses on the chemical reactions and optical density measurements. There is no mention of AI, ML, or related concepts.

Therapeutic

No
The device is an in vitro diagnostic (IVD) test used to aid in the diagnosis of Invasive Aspergillosis by detecting a specific antigen in patient samples. It does not directly treat or prevent a disease.

Diagnostic

Yes

The device is an "aid in the diagnosis of Invasive Aspergillosis" according to its intended use, which directly aligns with the definition of a diagnostic device.

SaMD (Software as a Medical Device)

No

The device is an immunoenzymatic assay kit, which is a physical collection of reagents and microplates used for laboratory testing. It is not software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states it's an "immunoenzymatic sandwich microplate assay for the detection of Aspergillus galactomannan antigen in adult and pediatric serum and Bronchoalveolar Lavage (BAL) fluid samples." This clearly indicates it's used to test samples taken from the human body to provide information about a person's health.
Aid in Diagnosis: The intended use also states it's used "as an aid in the diagnosis of Invasive Aspergillosis." This is a key characteristic of an IVD – providing information to assist in diagnosing a disease or condition.
Device Description: The description details a laboratory assay performed on human samples (serum and BAL fluid) using reagents and a microplate. This is consistent with the nature of an in vitro diagnostic test.
Performance Studies: The document includes detailed performance studies evaluating the device's sensitivity, specificity, and predictive values using human serum and BAL fluid samples from different patient populations. This type of performance evaluation is standard for IVDs to demonstrate their accuracy and reliability for their intended use.
Key Metrics: The listed key metrics (Sensitivity, Specificity, PPV, NPV, etc.) are all standard metrics used to evaluate the performance of diagnostic tests.
Predicate Device: The mention of a predicate device (K060641; Platelia™ Aspergillus EIA) indicates that this device is being compared to a previously cleared IVD, which is a common regulatory pathway for new IVDs.

All of these points strongly support the classification of this device as an In Vitro Diagnostic.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Platelia™ Aspergillus 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.

Product codes (comma separated list FDA assigned to the subject device)

NOM

Device Description

The Platelia™ Aspergillus EIA is a one-stage immunoenzymatic sandwich microplate assay which detects galactomannan in human serum and BAL fluid samples. The assay uses rat EBA-2 monoclonal antibodies, which are directed against Aspergillus galactomannan, and have been characterized in previous studies. The monoclonal antibodies are used, (1) to coat the wells of the microplate and bind the antigen, and (2) to detect the antigen bound to the sensitized microplate reagent: peroxidase-linked monoclonal antibodies).

Serum or BAL fluid samples are heat-treated in the presence of EDTA in order to dissociate immune complexes and to precipitate proteins that could possibly interfere with the test. The treated samples and conjugate are added to the wells coated with monoclonal antibodies, and incubated. A monoclonal antibody - galactomannan monoclonal antibody / peroxidase complex is formed in the presence of galactomannan antigen.

The strips are washed to remove any unbound material. Next, the substrate solution is added, which will react 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 620/630 nm wavelength.

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

adult and pediatric

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

Serum Samples:

Pediatric Patients:
- Sample Size: 1954 serum samples from 129 immunocompromised pediatric patients (Age

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.

Summary

0

.

:

·

510k Summary K093678

SECTION 2 510(k) SUMMARY

1

510k Summary K093678

510(k) SUMMARY:

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:
DATE:	November 20th, 2009

| APPLICANT: | Bio-Rad
3, Boulevard Raymond Poincar
92430 Marnes-la-Coquette, France |
|-------------------------|-----------------------------------------------------------------------------|
| OFFICIAL CORRESPONDENT: | Dr. Sylvie Confida |

TELEPHONE:	33-1-47-95-6138
FAX:	33-1-47-95-6242
PRODUCT TRADE NAME:	Bio-Rad Laboratories Platelia™ Aspergillus EIA
COMMON NAME:	Aspergillus Antigen EIA
CLASSIFICATION NAME:	Antigen, Galactomannan, Aspergillus spp.
PREDICATE DEVICE:	Platelia™ Aspergillus EIA

DEVICE DESCRIPTION

The Platelia™ Aspergillus EIA is a one-stage immunoenzymatic sandwich microplate assay which detects galactomannan in human serum and BAL fluid samples. The assay uses rat EBA-2 monoclonal antibodies, which are directed against Aspergillus galactomannan, and have been characterized in previous studies. The monoclonal antibodies are used, (1) to coat the wells of the microplate and bind the antigen, and (2) to detect the antigen bound to the sensitized microplate reagent: peroxidaselinked monoclonal antibodies).

Serum or BAL fluid samples are heat-treated in the presence of EDTA in order to dissociate immune complexes and to precipitate proteins that could possibly interfere with the test. The treated samples and conjugate are added to the wells coated with monoclonal antibodies, and incubated. A monoclonal antibody - galactomannan monoclonal antibody / peroxidase complex is formed in the presence of galactomannan antigen.

2

The strips are washed to remove any unbound material. Next, the substrate solution is added, which will react 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 620/630 nm wavelength.

INTENDED USE

The Platelia™ Aspergillus ElA 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.

INDICATIONS FOR USE

The Platelia™ Aspergillus 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.

TECHNOLOGICAL CHARACTERISTICS

The following tables summarize similarities and differences between the Platelia™ Aspergillus EIA (62793) and the current Platelia™ Aspergillus EIA (K060641):

Table 1(a) Similarities between intended use

| Similarities in
Function and Use | Platelia™ Aspergillus EIA,
Catalog 62793 | Platelia™ Aspergillus EIA
current (K060641) |
|-------------------------------------|---------------------------------------------|------------------------------------------------|
| Intended Use | Galactomannan antigen
detection. | Galactomannan antigen
detection. |

3

.

| Differences in
Function and Use | Platelia™ Aspergillus EIA,
Catalog 62793 | Platelia™ Aspergillus EIA
(K060641) |
|--------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------|
| Intended Use | Detection of Aspergillus
galactomannan antigen in
adult and pediatric serum and
Bronchoalveolar Lavage
(BAL) Fluid samples. | Detection of Aspergillus
galactomannan antigen in adult
and pediatric serum samples. |
| Matrix | Serum and Bronchoalveolar
Lavage (BAL) Fluid samples | Serum |
| Intended Use
/Indications for Use | Both the Intended Use and
Indications for Use are same. | The Intended Use and
Indications for Use are
different. |

Table 1(b) Differences between intended use

Table 2 Similarities between reagents and materials

| Similarities in
Components /
Materials | Platelia™ Aspergillus EIA,
Catalog 62793 | Platelia™ Aspergillus EIA
(K060641) |
|----------------------------------------------|-------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------|
| Microplate | 96 well microplate -
antibody coated microwells | 96 well microplate - antibody
coated microwells |
| Reagents | Conjugate, Wash Buffer,
Substrate, TMB Chromogen,
Sample Diluent, Positive
Control, Stop Solution. | Conjugate, Wash Buffer,
Substrate, TMB Chromogen,
Sample Diluent, Positive
Control, Stop Solution. |

Table 3 Similarities between assay procedures.

| Similarities in
Assay Procedures | Platelia™ Aspergillus EIA
Catalog 62793 | Platelia™ Aspergillus EIA
(K060641) |
|---------------------------------------------------------------------------------------------------------------|--------------------------------------------|----------------------------------------|
| Incubation
temperature of the
microplate after
addition of the
conjugate and the
treated sera. | Incubation temperature: 37°C | Incubation temperature: 37°C |
| Incubation time of
the microplate after
addition of the
conjugate and the
treated sera. | Incubation time: 90 ± 5
minutes | Incubation time: 90 ± 5
minutes |

4

| Differences in
Limitations of the
Procedure (Section
13) | Platelia™ Aspergillus EIA,
Catalog 62793 | Platelia™ Aspergillus EIA
(K060641) |
|-------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------|
| Limitation: Point 11 | Addition of Histoplasma and
Geotrichum to the list of
fungi causing cross-reactivity | |
| Limitation: Point 12 | Addition of a limitation that
cross-reactivity of BAL fluid
with Mycoplasma and
anaesthetic drugs used to
numb the neck/throat area
has not been evaluated. | |
| Limitation: Point 13 | Addition of limitation
regarding positive reactions
observed in patients
receiving products
containing sodium gluconate,
galactofuranose or
galactomannan. | |
| Limitation: Point 14 | Addition of limitation
regarding positive reactions
observed in patients
receiving PLASMA-
LYTE™ solution. | |
| Limitation: Point 15 | Addition of limitation about
assay results in BAL fluid
samples from
immunocompromised
patients to be interpreted
with caution. | |

Table 4 Differences between Limitations of the Procedure

5

Limitation: Point 16	Results of the Platelia™
	Aspergillus EIA in
	Bronchoalveolar Lavage
	(BAL) fluid samples between
	0.5-1.0 index have a lower
	predictive value than BAL
	sample results > 1.0 index
	values, therefore the results
	between 0.5-1.0 index values
	should be reviewed and
	supported by other clinical,
	radiological or laboratory
	evidence of invasive
	aspergillosis8, 17

PERFORMANCE EVALUATION SUMMARY

A. EXPECTED VALUES

I. SERUM

The expected prevalence of Invasive Aspergillosis varies with the patient population: rates from 5-20% have been reported.

The following results have been obtained from clinical studies conducted on pediatric (age 1.5. The y-axis is labeled "Number of Sera" and ranges from 0 to 900. The bar graph shows that the highest number of sera is in the 0.2-0.3 range, with a value of 805.

*Note: 80 samples, from 4 control patients with positive galactomannan antigen results coinciding with piperacillin/tazobactam (Zosyn®) therapy were excluded.

Pediatric Patients diagnosed with Invasive Aspergillosis

The scatter plot depicts galactomannan assay results for the 249 serum samples from 17 patients in this study diagnosed with proven or probable Invasive Aspergillosis as defined by EORTC/NIAID definitions. Not every serum sample from each patient is expected to be positive. The expected prevalence of Invasive Aspergillosis varies with the patient population; rates from 5-20% have been reported 10.23. The prevalence rate of this study was 13.6%.

Figure 2

Image /page/6/Figure/8 description: The image is a scatter plot titled "Pediatric Proven/ Probable Aspergillosis: Distribution of Index/ Pediatric Patient (N=17)". The x-axis is labeled "Patient Number" and ranges from 0 to 18. The y-axis is labeled "Index" and ranges from 0.0 to >2.0. The scatter plot shows the distribution of index values for each patient, with a horizontal line at 0.5.

7

B. Adults

A clinical study was conducted on a total of 1724 serum samples from 172 bone marrow transplant (BMT) and leukemic patients diagnosed with and without Invasive Aspergillosis, at three testing centers in North America to determine the performance characteristics of the Platelia™ Aspergillus EIA. The distribution of index values for these populations is represented in the following charts.

Adult Patients diagnosed without Invasive Aspergillosis (control population)

A total of 1262 serum samples obtained from 143 bone marrow transplant (BMT) and leukemic patients at three testing centers in North America were tested with the Platelia™ Aspergillus EIA test. The distribution of index values is shown in the following chart:

Image /page/7/Figure/6 description: The image shows the text "Figure 3" in a bold, serif font. The text is black against a white background. The number 3 is slightly smaller than the word "Figure".

Image /page/7/Figure/7 description: The image is a bar graph titled "Distribution of the Serum Index Value from the Adult Control Population (N=1262)". The x-axis is labeled "Index" and shows index ranges from 0-0.1 to >1.5. The y-axis is labeled "Number of Sera" and ranges from 0 to 800. The bar graph shows the number of sera for each index range, with the highest number of sera in the 0.2-0.3 range (662).

Adult Patients diagnosed with Invasive Aspergillosis

This scatter plot depicts galactomannan assay results for the 462 serum samples from 29 patients in this study diagnosed with proven or probable Invasive Aspergillosis as defined by EORTC/NIAID definitions. Not every serum sample from each patient is expected to be positive. The expected prevalence of Invasive Aspergillosis varies with the patient population; rates from 5-20% have been reported 10,23. The prevalence rate for this study was 16.9%.

8

Image /page/8/Figure/2 description: The image shows the words "Figure 4" in a serif font. The words are in bold and are black. There is a period after the number 4.

Image /page/8/Figure/3 description: The image is a scatter plot titled "Adult Proven/Probable Aspergillosis: Distribution of Index/Adult Patient (N=29)". The x-axis is labeled "Patient Number" and ranges from 0 to 30. The y-axis is labeled "Index" and ranges from 0.0 to greater than or equal to 2.0. The scatter plot shows the distribution of index values for each patient, with a horizontal line at 0.5.

The following graphs represent examples of a patient without clinical signs or symptoms óf Invasive Aspergillosis (negative for Aspergillus) and a patient with proven or probable Invasive Aspergillosis (positive for Aspergillus) respectively.

Image /page/8/Figure/5 description: This image contains the text "Figure 5 Negative patient". The text is in bold font. The figure number is at the top, and the description of the figure is below it.

Image /page/8/Figure/6 description: The image is a graph titled "CONTROL PATIENT". The x-axis is labeled "Days" and ranges from 0 to 60. The y-axis is labeled "Index" and ranges from 0 to 1.6. The graph shows a line with diamond markers that fluctuates between 0 and 0.3, with a few peaks around 0.2, and a horizontal line at 0.5.

9

Figure 6 Positive patient

Image /page/9/Figure/3 description: This image is a line graph titled "PROVEN INVASIVE ASPERGILLOSIS PATIENT". The x-axis is labeled "Days" and ranges from 0 to 60. The y-axis is labeled "Index" and ranges from 0 to 1.6. The line on the graph starts at approximately 0.2, rises sharply around day 25, peaks around day 30 at approximately 1.5, and then gradually declines to around 0.4 by day 55.

II. BAL FLUID

Two studies were conducted on a total of 449 BAL samples from 178 Solid Organ transplant (SOT) and lung transplant recipients with and without invasive aspergillosis in the United States to determine the performance characteristics of the Platelia™ Aspergillus EIA kit with Bronchoalveolar Lavage Fluid samples.

Of these, there were 403 BAL samples from 167 solid organ and lung transplant recipients without invasive aspergillosis.

In addition, a retrospective analysis was performed on BAL samples from 99 evaluable high risk haematology patients in a study outside the United States which included 58 patients with proven or probable invasive aspergillosis.

Expected values in BAL samples from the combined SOT and lung transplant recipients without Invasive Aspergillosis are presented in the table below. Results are presented by samples from transplant recipients with and without mold colonization.

Table 1

Expected Values by Sample Combined SOT and Lung Transplant Recipients without Invasive Aspergillosis N =403 BAL Fluids

Diagnosis	N	Positive (%)	Negative (%)
Controls without
colonization	341	11/341(3.2%)	330/341(96.8%)
Controls with colonization	62	12/62
(19.4%)	50/62 (80.6%)
Control Total	403	23/403(5.7%)	380/403(94.3%)

Expected values in BAL samples from the combined SOT and lung transplant recipients without Invasive Aspergillosis are presented by transplant type in the table below.

10

Table 2

Expected Values by Sample Combined SOT and Lung Transplant Recipients without Invasive Aspergillosis By Transplant Type N =403 RAL Fluids

N - 403 BAL Fluids
Transplant Type	N	Positive (%)	Negative (%)
Heart	28	3/28 (10.7%)	25/28 (89.3%)
Kidney	25	3/25 (12.0%)	22/25 (88.0%)
Liver	23	1/23 (4.3%)	22/23 (95.7%)
Lung	327	16/327 (4.9%)	311/327
(95.1%)
Control Total	403	23/403
(5.7%)	380/403
(94.3%)

Expected values were also evaluated in a total of 41 BAL fluid samples from 41 hematological disease patients without Invasive Aspergillosis and are presented in the Table below

Table 3

Expected Values by Sample Hematologic disease patients without Invasive Aspergillosis N =41 BAL Fluids

| Alleria Million Art Program And any Art All All All All All All All All All All All All All All All All All All All All All All All All All All All All All All All All All Al
AND AND A B A A Libray on Street States
Diagnosis | ALL House of the American | "Whileser deberg for working with Address of the Address of the Artistics of Articles of Articles of Articles of Articles of Articles of Articles of Articles of Articles of A
Positive (%) | Negative (%) |
|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------|
| Controls | | 8/41 (19.5%) | 33/41 (80.5%) |

B. REPRODUCIBILITY STUDIES

a) Reproducibility Studies In Serum

Inter-assay and Intra-assay variability for the Platelia™ Aspergillus EIA were determined in a study using a panel of 6 pooled patient serum samples (one negative, one low positive, two positive, and two high positive) obtained at three clinical trial sites in North America. Each of the 6 panel members were tested in triplicate (x3) on 3 different days, on one lot, at two sites (total number of replicates at each site = 9). Each of the 6 panel members was tested in duplicate (x2) on 3 different days, on 1 lot, at a third site (total number of replicates at the third site = 6). One (1) operator performed all precision testing at each site. The data were analyzed according to the Clinical Laboratory Standards Institute (CLSI) (formerly National Committee for Clinical Laboratory Standards (NCCLS)). The mean optical density (OD) and mean index value, standard deviation (SD), percent coefficient of variation (%CV), within run precision (intraassay) and within site (inter-assay) precision for each panel member at each site are illustrated below in the following tables.

11

Table 4

C:
1

Site 1	Panel Member	Neg		Low Pos		Pos #1		Pos #2		High Pos#1		High Pos #2		Neg Control		CO Control		Pos Control
		OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index
	N	9	9	9	9	9	9	9	9	9	9	9	9	3	3	6	6	3	3
	Mean	0.052	0.09	0.445	0.74	0.702	1.17	0.931	1.563	1.227	2.06	2.887	4.83	0.046	0.08	0.606	1.00	2.216	3.67
	Within Run
(intra-assay)1 SD	0.002	0.00	0.022	0.03	0.059	0.09	0.044	0.08	0.051	0.09	0.089	0.17	N/A	N/A	0.02	0.03	N/A	N/A
	%CV	N/A	N/A	4.8%	4.4%	8.4%	7.6%	4.7%	5.1%	4.2%	4.4%	3.1%	3.6%	N/A	N/A	3.7%	3.4%	N/A	N/A
	Total
(inter-assay)2 SD	0.036	0.04	0.051	0.08	0.070	0.14	0.044	0.25	0.058	0.29	0.169	0.58	N/A	N/A	0.102	0.03	0.317	0.12
	%CV	N/A	N/A	11.5%	10.4%	10.0%	11.6%	4.7%	15.7%	4.7%	14.3%	5.9%	11.9%	N/A	N/A	16.9%	2.8%	14.3%	3.3%
Site 2	Panel Member	Neg		Low Pos		Pos #1		Pos #2		High Pos#1		High Pos #2		Neg Control		CO Control		Pos Control
		OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index
	N	9	9	9	9	9	9	9	9	9	9	9	9	3	3	6	6	3	3
	Mean	0.040	0.10	0.280	0.70	0.364	0.89	0.602	1.49	0.801	2.01	1.381	3.43	0.074	0.18	0.415	1.00	1.197	2.97
	Within Run
(intra-assay)1 SD	0.006	0.01	0.041	0.09	0.023	0.07	0.045	0.11	0.046	0.10	0.047	0.11	N/A	N/A	0.00	0.01	N/A	N/A
	%CV	N/A	N/A	14.5%	13.0%	6.4%	7.6%	7.5%	7.1%	5.7%	4.8%	3.5%	3.2%	N/A	N/A	1.1%	1.1%	N/A	N/A
	Total
(inter-assay)2 SD	0.006	0.03	0.058	0.19	0.083	0.18	0.057	0.28	0.042	0.53	0.079	1.00	N/A	N/A	0.094	0.01	0.068	0.54
	%CV	N/A	N/A	20.8%	27.0%	22.7%	19.8%	9.5%	18.7%	5.3%	26.5%	5.8%	29.2%	N/A	N/A	22.7%	0.9%	5.7%	18.2%
Site 3	Panel Member	Neg		Low Pos		Pos #1		Pos #2		High Pos#1		High Pos #2		Neg Control		CO Control		Pos Control
		OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index	OD	Index
	N	6	6	6	6	6	6	6	6	6	6	6	6	3	3	6	6	3	3
	Mean	0.049	0.10	0.388	0.81	0.652	1.36	0.830	1.73	1.158	2.41	2.378	4.96	0.059	0.12	0.480	1.00	1.652	3.45
	Within Run
(intra-assay)1 SD	0.003	0.01	0.009	0.02	0.082	0.17	0.068	0.14	0.094	0.20	0.126	0.25	N/A	N/A	0.028	0.06	N/A	N/A
	%CV	N/A	N/A	2.4%	2.4%	12.5%	12.2%	8.2%	8.2%	8.1%	8.2%	5.3%	5.1%	N/A	N/A	5.8%	5.8%	N/A	N/A
	Total
(inter-assay)2 SD	0.012	0.03	0.078	0.13	0.068	0.15	0.104	0.25	0.082	0.15	0.111	0.34	N/A	N/A	0.028	0.04	0.056	0.23
	%CV	N/A	N/A	20.0%	15.8%	10.5%	11.1%	12.5%	14.3%	7.1%	6.2%	4.7%	6.8%	N/A	N/A	5.8%	4.1%	3.4%	6.6%

N/A = not applicable

'NCCLS EP5-A, Vol. 19, No. 2, Page 24, Equation (C2) 2 NCCLS EP5-A, Vol. 19, No. 2, Page 25, Equation (C3) and Equation (C4)

b) Reproducibillity Studies in BAL

Inter-assay and Intra-assay variability for the Platelia™ Aspergillus EIA were determined in a study using a panel of 4 pooled patient BAL samples spiked with purified galactomannan (one negative, one high negative, one low positive and one medium positive) at 3 testing sites (Two US clinical testing sites and one internal site). Each of the 4 panel members and the controls were tested in duplicate (x2) in 2 runs per day on 5 different days on one lot (Total number of replicates at each site = 120). Two (2) operators performed all precision testing at each site. The data was analyzed according to the Clinical Laboratory Standards Institute (CLSI) (formerly National Committee for Clinical Laboratory Standards (NCCLS)). The mean optical density (OD) and mean index value, standard deviation (SD), percent coefficient of variation (%CV), within run precision (intra-assay) and between site, between day, between operator and between run (inter-assay) precision for each panel member are illustrated below in the following table:

12

Table 5

Combined Sites Summary

| Summary | | Negative
N= 60 | | High Negative
N=60 | | Low
Positive
N=60 | | Medium
Positive
N=60 | | Positive
Control
N=60 | | Negative
Control
N=60 | |
|------------------|-----|-------------------|-------|-----------------------|-------|-------------------------|-------|----------------------------|-------|-----------------------------|-------|-----------------------------|-------|
| | | OD | Index | OD | Index | OD | Index | OD | Index | OD | Index | OD | Index |
| Mean | | 0.121 | 0.29 | 0.214 | 0.50 | 0.375 | 0.88 | 0.575 | 1.35 | 1.580 | 3.72 | 0.047 | 0.11 |
| Within
Run | SD | N/A | N/A | 0.037 | 0.103 | 0.035 | 0.078 | 0.029 | 0.067 | 0.111 | 0.265 | N/A | N/A |
| (Intra
Assay) | %CV | N/A | N/A | 17.4% | 20.5% | 9.3% | 8.9% | 5.0% | 5.0% | 7.0% | 7.1% | N/A | N/A |
| Total | SD | N/A | N/A | 0.042 | 0.095 | 0.061 | 0.122 | 0.070 | 0.138 | 0.190 | 0.438 | N/A | N/A |
| (Inter
Assay) | %CV | N /A | N/A | 19.6% | 18.9% | 16.2% | 13.9% | 12.2% | 10.2% | 12.0% | 11.8% | N/A | N/A |

C. PERFORMANCE EVALUATION STUDIES

I. SERUM SAMPLES

Clinical testing to evaluate the sensitivity, specificity, and predictive value of the Platelia™ Aspergillus EIA was conducted on pediatric (age ≤ 21 years) patients at three sites located in the United States and on adult patients at three sites located in North America. The studies were conducted using a total of 1954 serum samples collected from 129 pediatric patients and a total of 1724 serum samples collected from 172 adult patients from the following populations*:

· patients without signs of Invasive Aspergillosis (control patients)

· patients with Probable Invasive Aspergillosis

· patients with Proven Invasive Aspergillosis

The Invasive Fungal Infection Cooperative Group (IFICG) of the European Organization for Research and Treatment of Cancer (EORTC) and the Mycosis Study Group (MSG) of the National Institute of Allergy and Infectious Diseases (NIAID) in 2002 have defined criteria for diagnosis of Invasive Aspergillosis (IA) in patients with hematologic malignancy or hematopoetic stem cell transplant. 4

SENSITIVITY

A. Pediatrics

Results from this study have been analyzed in terms of patient sensitivity. Sensitivity testing was conducted using the Platelia™ Aspergillus EIA at three sites on a combined total of 17 immunocompromised pediatric patients diagnosed with Proven or Probable Invasive Aspergillosis.

13

Table 6

| Diagnosis | Number of
patients | Sensitivity | 95% Confidence
Interval |
|-----------------------------------------------|-----------------------|--------------|----------------------------|
| Proven Aspergillosis | 9 | 44.4% (4/9) | 18.9-73.3% |
| Probable Aspergillosis | 8 | 62.5% (5/8) | 30.6-86.3% |
| Combined Proven and
Probable Aspergillosis | 17* | 52.9% (9/17) | 31.0-73.8% |

*Note: 8 of the 17 patients gave negative Aspergillus galactomannan antigen results. All of the 8 patients with negative Aspergillus galactomannan antigen results received therapy with multiple antifungal agents. The concomitant use of mold-active anti-fungal therapy in some patients with Invasive Aspergillosis may result in reduced sensitivity 1.

B. Adults

Sensitivity testing was conducted using the Platelia™ Aspergillus ElA at three sites on a combined total of 29 Bone Marrow Transplant (BMT) and Leukemia adult patients diagnosed with Proven or Probable Invasive Aspergillosis.

Table 7

| Diagnosis | Number of
patients | Sensitivity | 95% Confidence
Interval |
|-----------------------------------------------|-----------------------|---------------|----------------------------|
| Proven Aspergillosis | 11 | 81.8% (9/11) | 52.3-94.9% |
| Probable Aspergillosis | 18 | 77.8% (14/18) | 54.8-91.0% |
| Combined Proven and
Probable Aspergillosis | 29 | 79.3% (23/29) | 61.6-90.2% |

SPECIFICITY

A. Pediatrics

Specificity by pediatric patients

Specificity testing was conducted using the Platelia™ Aspergillus EIA at three sites on a combined total of 108* immunocrompomised pediatric patients without signs of Invasive Aspergillosis (control patients).

Table 8

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| Site | Number of patients | Specificity | 95% Confidence
Interval |
|----------------|--------------------|----------------|----------------------------|
| 1 | 44 | 86.4 % (38/44) | 73.3-93.6% |
| 2 | 59 | 86.4 % (51/59) | 75.5-93.0% |
| 3 | 5 | 100% (5/5) | 56.6-100% |
| Combined Sites | 108 | 87.0% (94/108) | 79.4-92.1% |

*Note: 4 patients with positive galactomannan antigen results coinciding with piperacillin / tazobactam therapy were excluded.

Specificity by pediatric samples

Specificity testing was conducted using the Platelia™ Aspergillus EIA at three sites on a combined total of 1625* samples obtained from 108 immunocrompomised pediatric patients without signs of Invasive Aspergillosis (control patients).

| Site | Number of samples | Specificity | 95% Confidence
Interval |
|----------------|-------------------|-------------------|----------------------------|
| 1 | 794 | 98.9% (785/794) | 97.9-99.4% |
| 2 | 731 | 97.8% (715/731) | 96.5-98.6% |
| 3 | 100 | 100% (100/100) | 96.3-100% |
| Combined Sites | 1625 | 98.5% (1600/1625) | 97.7-99.0% |

Table 9

*Note: 80 samples from 4 patients with positive galactomannan antigen results coinciding with piperacillin / tazobactam therapy were excluded.

B. Adults

Specificity by adult patients

Specificity testing was conducted using the Platelia™ Aspergillus EIA at three sites on a combined total of 143 Bone Marrow Transplant (BMT) and Leukemia adult patients without signs of Invasive Aspergillosis (control patients).

15

| Site | Number of patients | Specificity | 95% Confidence
Interval |
|----------------|--------------------|-----------------|----------------------------|
| 1 | 28 | 78.6% (22/28) | 60.5-89.8% |
| 2 | 77 | 93.4% (71/77) | 84.0-96.4% |
| 3 | 38 | 89.5% (34/38) | 75.9-95.8% |
| Combined Sites | 143 | 88.8% (127/143) | 82.6-93.0% |

Specificity by adult samples

Specificity testing was conducted using the Platelia™ Aspergillus EIA at three sites on a combined total of 1262 samples obtained from 143 Bone Marrow Transplant (BMT) and Leukemia adult patients without signs of Invasive Aspergillosis (control patients).

| 'able
1
I

T
----------------------

| Site | Number of samples | Specificity | 95% Confidence
Interval |
|----------------|-------------------|-------------------|----------------------------|
| 1 | 349 | 98.0% (342/349) | 95.9-99.0% |
| 2 | 560 | 98.6% (552/560) | 97.2-99.3% |
| 3 | 353 | 98.9% (349/353) | 97.1-99.6% |
| Combined Sites | 1262 | 98.5% (1243/1262) | 97.7-99.0% |

PREDICTIVE VALUE

Positive and negative predictive values have been analyzed for the patient population in this study. Based on the actual average of 13.6% prevalence rate in pediatrics and 16.9% prevalence rate in adults observed in this study, positive and negative predictive values have been calculated as below:

A. Pediatrics

Study Prevalence 13.6%
PPV: 39.1%	95% Confidence Interval : 22.2-59.2%
NPV: 92.2%	95% Confidence Interval : 85.3-96.0%

B. Adults

Study Prevalence 16.9%
PPV: 59.0%	95% Confidence Interval:43.4-72.9%
NPV: 95.5%	95% Confidence Interval: 90.5-97.9%

The expected prevalence of Invasive Aspergillosis varies with the patient population; rates from 5-20% have been reported 19,23. For patient populations on the lower end of the

16

510k Summary K093678

published prevalence, the positive and negative predictive values have been re-calculated using a 5% prevalence rate.

A. Pediatrics

Calculated Prevalence 5%
PPV : 17.6%	95% Confidence Interval : 6.5-39.8%
NPV : 97.2%	95% Confidence Interval : 92.1-99.1%

B. Adults

Calculated Prevalence 5%
PPV: 27.2%	95% Confidence interval: 13.7-46.7%
NPV: 98.8%	95% Confidence Interval: 95.4-99.7%

CROSS REACTIVITY

a) Cross Reactivity - Serum

A study to evaluate the effect of potentially interfering medical conditions unrelated to Invasive Aspergillosis was performed with one lot of the Platelia™ Aspergillus EIA kit. The following serum samples were tested for cross-reactivity with the PlateliaTM Aspergillus EIA. A total of 151 sera were tested.

Table 12

. . . . . .

Pathology	# Samples Tested	# Positives
Rheumatoid Factor	10	0
ANA Positive	10	0
IgG Hypergammaglobulinemia	10	0
IgM Hypergammaglobulinemia	10	0
Cancer*	11	0
Non-Viral Cirrhosis (primary biliary;
alcohol induced; drug induced)	10	0
Multiple Transfusions	10	0
Multiparous Females	10	0
HAV	10	0
HCV	10	0
Rubella	10	0
CMV	10	0
Syphilis (RPR+)	10	0
Toxoplasmosis	10	0
Mycoplasma	10	0

One each of bladder, breast(2), colon, endometrial, lung, prostate, renal, and squamous(3).

II. BAL FLUID SAMPLES- Performance Characteristics

Sensitivity and specificity of the Platelia™ Aspergillus EIA with BAL fluid samples were evaluated in two studies in the United States on 116 samples from 62 solid organ transplant recipients and 333 samples from 116 lung transplant recipients and one study

17

outside the United States on 99 samples from 99 high risk hematology patients with and without invasive aspergillosis.

A. SENSITIVITY

Sensitivity was evaluated in Solid Organ Transplant and Lung Transplant recipients diagnosed with invasive aspergillosis as well as hematologic disease patients diagnosed with invasive aspergillosis according to the EORTC/MSG criteria.

I. Solid Organ Transplant recipients with Invasive Aspergillosis

Of the total of 116 samples from 62 Solid Organ Transplant recipients in one study, sensitivity was evaluated in 5 recipients diagnosed with invasive aspergillosis as shown in the table below.

Table 13

Proven or Probable Invasive Aspergillosis in Solid Organ Transplant Recipients

| Diagnosis | N | Index ≥ 0.5 | Sensitivity | 95% Confidence
Interval |
|--------------------------------------------------|---|-------------|-------------|----------------------------|
| Proven Aspergillosis | 2 | 2 | 2/2 (100%) | 34.2 - 100% |
| Probable
Aspergillosis | 3 | 3 | 3/3 (100%) | 43.8 - 100% |
| Combined Proven
and Probable
Aspergillosis | 5 | 5 | 5/5 (100%) | 56.5 - 100% |

By Patient

Table 14

Proven or Probable Invasive Aspergillosis in Solid Organ Transplant Recipients By Transplant Type

| Transplant
Type | N | Index ≥
0.5 | Sensitivity | 95%
Confidence
Interval |
|--------------------|---|----------------|-------------|-------------------------------|
| Heart | 1 | 1 | 1/1 (100%) | 20.6 - 100% |
| Kidney | 3 | 3 | 3/3 (100%) | 43.8 - 100% |
| Liver | 1 | 1 | 1/1 (100%) | 20.6 - 100% |
| Total | 5 | 5 | 5/5 (100%) | 56.5 - 100% |

18

II. Lung Transplant recipients with invasive aspergillosis

Of the total of 333 samples from 116 Lung Transplant recipients in another study, sensitivity was evaluated in 6 recipients diagnosed with invasive aspergillosis as shown in the table below.

Table 15

Proven or Probable Invasive Aspergillosis in Lung Transplant Recipients By patient

| Diagnosis | N | Index ≥ 0.5 | Sensitivity | 95%
Confidence
Interval |
|-----------------------------------------------|---|-------------|-------------|-------------------------------|
| Proven Aspergillosis | 2 | 1 | 1/2 (50.0%) | 9.4 - 90.6% |
| Probable Aspergillosis | 4 | 3 | 3/4 (75.0%) | 30.0 - 95.4% |
| Combined Proven and
Probable Aspergillosis | 6 | 4 | 4/6 (66.7%) | 30.0 - 90.3% |

III. Hematologic disease patients with invasive aspergillosis

Sensitivity was also evaluated in a third study in 58 samples from 58 hematologic disease patients diagnosed with invasive aspergillosis as shown in the table below. In the study a retrospective analysis was performed on BAL samples from high risk hematology patients using the Platelia™ Aspergillus EIA. The data from this published study below was evaluated to establish the performance characteristics of the Platelia™ Aspergillus EIA on BAL fluid.

Maertens et al. 2009 Bronchoalveolar Lavage Fluid Galactomannan for the Diagnosis of Invasive Pulmonary Aspergillosis in Patients with Hematologic Diseases. Clin. Infec. Diseas.49:1688-93

Table 16

| Diagnosis | N | Index ≥ 0.5 | Sensitivity | 95%
Confidence
Interval |
|-----------------------------------------------|----|-------------|------------------|-------------------------------|
| Proven Aspergillosis | 31 | 31 | 31/31 (100%) | 89.0 - 100% |
| Probable Aspergillosis | 27 | 26 | 26/27 (96.3%) | 81.7 - 99.3% |
| Combined Proven and
Probable Aspergillosis | 58 | 57 | 57/58
(98.3%) | 90.8 - 99.7% |

Proven or Probable Invasive Aspergillosis in Hematologic Disease Patients

19

B. SPECIFICITY

Specificity was evaluated in a total of 98 BAL samples from 57 SOT recipients and 305 BAL samples from 110 Lung Transplant recipients without invasive aspergillosis and is shown in the table below. Results are presented by samples from transplant recipients with and without mold colonization.

Table 17

Specificity by Sample Combined SOT and Lung Transplant Recipients without Invasive Aspergillosis N = 403 BAL Fluids

| Diagnosis | N | Index