The Accuwell" 17α-Hydroxyprogesterone EIA Kit is designed for the quantitative measurement of 17α-Hydroxyprogesterone (17-OHP) concentrations in neonatal blood samples that have been collected onto Whatman 903® specimen collection paper. The results are used to screen newborns for classical congenital adrenal hyperplasia (CAH).

Device Description

The Accuwell" 17a-Hydroxyprogesterone EIA Kit is a competitive solid phase enzyme immunoassay (ElA) for the quantitative measurement of 17«-Hydroxyprogesterone (17-OHP) concentrations in neonatal blood samples that have been collected onto Whatman S&S 903" specimen collection paper. Standard control, and unknown dried blood spot sample discs are added to specified wells in a 90-well microplate coated with antiserum specific for 17-OHP.

AI/ML Overview

Here's an analysis of the acceptance criteria and supporting study for the Accuwell 17-alpha-Hydroxyprogesterone EIA, based on the provided text:

Device: Accuwell 17-alpha-Hydroxyprogesterone (OHP) enzyme immunoassay (EIA) for neonatal screening.

Acceptance Criteria and Reported Device Performance

The acceptance criteria for this device are primarily demonstrated through comparison to a legally marketed predicate device (AutoDELFIA Neonatal 17a-OH-progesterone) and adherence to established internal precision and analytical sensitivity targets. While explicit "acceptance criteria" in terms of pass/fail thresholds for each metric are not always numerically stated as such, the document presents comparative data and targets that would have been used to demonstrate substantial equivalence.

Acceptance Criteria Category	Specific Metric	Acceptance Criteria (Implicit/Explicit)	Reported Device Performance (Accuwell 17-OHP EIA)
Intended Use Equivalence	Intended Use	To measure 17α-hydroxyprogesterone (17-OHP) levels in neonatal dried blood spots for screening newborns for classical congenital adrenal hyperplasia (CAH). (Identical to predicate)	Measures 17α-hydroxyprogesterone (17-OHP) concentrations in neonatal blood samples collected on Whatman 903® paper, used to screen newborns for classical CAH. (Meets)
Analytical Sensitivity	Limit of Detection	"Should be confirmed by each laboratory and appropriate concentrations defined." (Implies a low detection limit is desirable and comparable to predicate's 1.5 ng/ml)	Overnight Eluate: 2.2 ng/ml3 Hour Eluate: 1.5 ng/mlOvernight Direct: 2.4 ng/ml
Precision	Within-Run Precision (%CV)	Predicate: 8.8-11.3 (low), 8.9 (medium), 8.9 (high) %CVAccuwell: 3.5% (low), 3.7% (medium), 2.7% (high) %CV for overall 'Proposed Device' table. More detailed data below from dedicated studies.	From Dedicated Precision Study:ON Eluate: 10.7-11.4 %CV (total), within-run Sr provided3 HR Eluate: 11.3-12.9 %CV (total), within-run Sr providedON Direct DBS: 10.0-14.1 %CV (total), within-run Sr provided
	Between-Run Precision (%CV)	Predicate: 4.0-3.3 (medium) %CVAccuwell: 10.3-8.0 (medium), 11.2 (high) %CV for overall 'Proposed Device' table. More detailed data below from dedicated studies.	From Dedicated Precision Study:ON Eluate: Between-day Sdd provided3 HR Eluate: Between-day Sdd providedON Direct DBS: Between-day Sdd provided
	Total Precision (%CV)	Predicate: 12.0-9.5 (medium), 9.2 (high) %CVAccuwell: 11.0-8.9 (medium), 11.6 (high) %CV for overall 'Proposed Device' table. More detailed data below from dedicated studies.	From Dedicated Precision Study:ON Eluate: 10.7 (28.2 ng/ml), 8.8 (56.1 ng/ml), 11.4 (108.1 ng/ml) %CV3 HR Eluate: 12.9 (26.2 ng/ml), 7.7 (53.0 ng/ml), 11.3 (104.6 ng/ml) %CVON Direct DBS: 14.1 (29.5 ng/ml), 10.0 (57.9 ng/ml), 12.2 (108.6 ng/ml) %CV
Linearity & Recovery	Range	Approximately 2.0 ng/ml to 250 ng/ml.	Linear range and measuring range is approximately 2.0 ng/ml to 250 ng/ml. Measured % Recovery between 85.4% and 119.6% across 12 concentration levels (10-300 ng/ml). Regression R^2 values: 0.9994 (ON Eluate), 0.992 (3 Hr Eluate), 0.99 (ON Direct DBS).
Method Comparison	Correlation with Predicate (R-value, Slope, Intercept)	No explicit threshold given. Implied to be strong correlation (e.g., R > 0.9, slope near 1, intercept near 0) demonstrating substantial equivalence.	R-values consistently > 0.90 across all populations and methods (range 0.8421 to 0.9785). Slopes generally between 0.970 and 1.234. Intercepts between +1.081 and +6.569. (Specific values vary by population, method, percentile).
Specificity	Cross-Reactivity	List of compounds with cross-reactivity < 0.01% (many listed) and a few with higher (e.g., 21-Desoxycortisol 2.4%, 16a-Hydroxyprogestrone 1.2%, Progesterone 0.5%). This is an inherent characteristic, not typically a pass/fail criterion unless exceeding clinical acceptability.	Presented in table (e.g., 21-Desoxycortisol: 2.4%, Progesterone: 0.5%, Cholesterol: < 0.01%). Meets.
	Interfering Substances	"No detectable interference" for hemoglobin and bilirubin (conjugated/unconjugated). "Significant interference" from lipid solutions. (This indicates awareness of limitations and informs labeling).	Hemoglobin and bilirubin caused no detectable interference. Significant interference from lipid solutions. (Meets, with transparency on limitations).
Expected Values	Normal Ranges	Establish own normal range and cutoff levels due to population differences. Provided ranges are "example only" and "guidance."	Provided example "normal" and "follow-up" ranges for various populations (term/premature, age ranges) for each Accuwell method (ON Direct, ON Eluate, 3Hr Eluate). (Meets requirement to provide illustrative data).

Study Details

Sample size used for the test set and the data provenance:
- Method Comparison Study (Test Set):
  - Population >2500g b.w., 0-1 days old: 133 infant blood spot specimens.
  - Population >2500g b.w., 2-3 days old: 133 infant blood spot specimens.
  - Population >2500g b.w., >4 days old: 197 infant blood spot specimens.
  - Population 1400-2500g b.w., 1-6 days old: 30 infant blood spot specimens.
  - Total Samples for Method Comparison: 133 + 133 + 197 + 30 = 493 infant blood spot specimens.
  - Data Provenance: Retrospective study. Test samples were blinded neonatal dried blood spots collected in sequence under routine screening conditions from a U.S. department of public health laboratory. Original screening results for each sample using the predicate test kit were also obtained from the submitting laboratory.
- Precision Study:
  - Test samples: CDC Control Lots 451, 452, and 453 (enriched with 25, 50, and 100 ng/ml serum equivalents, respectively). These are laboratory control materials, not patient samples.
  - Number of measurements: 40 measurements for each control level across each of the three assay procedures (ON Eluate, 3 HR Eluate, ON Direct DBS).
- Analytical Sensitivity Study:
  - Test Samples: Zero standard (N=26).
  - Number of measurements: 26 measurements in one assay for each of the two procedures (eluate and direct, spanning elution times).
- Linearity and Recovery Study:
  - Test Samples: Dried blood spots representing 12 concentrations (10, 24.5, 39, 68, 97, 126, 155, 184, 213, 242, 271, 300 ng/ml).
  - Number of measurements per sample: Four aliquots of each of the 12 samples were tested in each of two runs using each of the three assay procedures. This implies 8 measurements per sample for each procedure.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- Not applicable (N/A) in the AI sense. This is an in vitro diagnostic device measuring a biomarker. "Ground truth" for the test set in the method comparison study was established by the measurement of these same samples using the predicate device (AutoDELFIA Neonatal 17a-OH-progesterone kit) in a reference lab setting, combined with the fact they were routine screening samples from a public health lab. The device's performance is gauged against another validated assay, not against expert human interpretations of images or clinical cases.
Adjudication method (e.g., 2+1, 3+1, none) for the test set:
- None. This is a quantitative assay, comparing the numerical output of the new device against the numerical output of a predicate device. There is no human diagnostic interpretation to adjudicate.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No. This is an in vitro diagnostic device for laboratory measurement, not an AI-assisted diagnostic imaging or interpretation tool for human readers. Therefore, an MRMC study is not applicable.
If a standalone (i.e. algorithm only without human-in-the loop performance) was done:
- Yes, effectively. The Accuwell 17-OHP EIA kit itself is a standalone diagnostic tool. Its performance (accuracy, precision, linearity, sensitivity, specificity) is evaluated independently, and then its results are compared to those of another "standalone" diagnostic kit (the predicate). While a human technician operates the EIA and reads the microplate spectrophotometer, the assay itself generates the quantitative results without human cognitive interpretation of subjective data. The "human-in-the-loop" here is the lab professional following the procedural steps.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Comparison to a legally marketed predicate device. In the context of 510(k) submissions for in vitro diagnostics, the predicate device's performance often serves as the "ground truth" for demonstrating substantial equivalence. The predicate device itself would have been validated against some form of clinical or analytical ground truth (e.g., confirmed CAH diagnoses, reference methods like GC-MS, or clinical outcomes), but for this submission, the predicate is the primary comparison.
The sample size for the training set:
- Not applicable (N/A) in the AI/machine learning sense. This is a chemical immunoassay kit, not a machine learning model that undergoes "training" on a dataset. The development of the assay would have involved various experimental iterations and optimizations, but there isn't a "training set" like in AI.
How the ground truth for the training set was established:
- Not applicable (N/A). As stated above, this is an immunoassay kit, not an AI model requiring a training set with established ground truth. The "training" of the device is through its manufacturing process and quality control, ensuring it produces accurate biochemical measurements based on validated reagents and procedures.

Summary

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Accuwell 17-alpha-Hydroxyprogesterone EIA 510(k) Summary

1) Submitter Name:	Genesis Northwest (dba) Neo-Genesis
Address:	15140 S.E. 82 Drive, Suite 270 Clackamas, OR 97015 USA
Telephone Number:	503-657-8000
Contact Person:	Jannet Baldwin, President
Date Prepared:	02/22/2007

K060452

MAR 13 2007

2) Device Trade or Proprietary Name:	Accuwell 17-a-hydroxyprogesterone (OHP) enzyme immunoassay (EIA)
Device Common or Usual Name:	17-a-hydroxyprogesterone (OHP) neonatal screening enzyme immunoassay (EIA)
Device Classification Name:	Radioimmunoassay, 17-hydroxyprogesterone (JLX)
3) Legally Marketed Predicate Device:	AutoDELFIA Neonatal 17a-OH-progesterone, Model B048-112

4 ) Device Description:

Summary and Explanation of the Assay

Congenital Adrenal Hyperplasia (CAH) is a family of genetic disorders caused by reduced enzyme activities, which are required in the biosynthesis pathway leading to the production of cortisol in the adrenal cortex. The most common of these disorders is the 21-hydroxylase deficiency and accounts for more than 90% of CAH cases. 1, 2, 3

The incidence of classical 21-hydroxylase deficiencies widely by population and efhnic group. In Japan the incidence is thought to be about 1 in 21,000, while in Europe and North America incidence ranges between 1 in 10,000 to 1 in 16,000. The Yupik Eskimos in Alaska have an extremely high incidence of around 1 in 300 4,5

Clinical symptoms of the 21-hydroxylase deficiency occur primarily due to the over production of precursors to the blocked enzymatic step. These precursors are shunted into the androgen biosynthesis pathway, which produces virilization in the female fetus, rapid postnatal growth with accelerated skeletal maturation, precocious puberty and short stature in both males and females. About 75% of the patients with classical CAH also have a defect in their ability to synthesize aldosterone, which can lead to a salt wasting crisis.

In classical 21-hydroxylase deficiencies the 17 α-hydroxyprogesterone (17-OHP) levels are markedly increased. However, the 17-OHP levels in normal infants are elevated in the first two to three days after birth. These high concentrations in normal infants decline rapidly to adult levels within the first seven to ten days.' Low birth-weight, premature and sick babies may continue to have higher than normal 17-OHP levels for an extended time frame. For this reason it has been recommended that a multi-tiered approach

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be employed to setting the normal cut-off. Laboratories must be notified of either dexamethasone therapy or transfusion status. Due to the population differences it is also strongly recommended that each laboratory establish their own normal range and cutoff levels. Periodic review and possible adjustment if necessary to reduce the false positive rate is also recommended. Non-classical forms of CAH are not
reliably detected by newborn screening." 9.10.11

The Working Group on Neonatal Screening of the European Society of Pediatric Endocrinology (ESPE) has recommended that after a positive screening test for CAH, it is imperative that the diagnosis be validated with confirmatory testing. Confirmation methods vary depending on availability but include urine steroid analysis, ACTH stimulation and CY21 gene analysis.12, 13, 1

Principle of the Assay

Two procedures are presented.

The first procedure includes an elution step where blood is eluted from the dried blood spot sample. A 1/8 inch disc from standards, controls, and unknown dried blood spot samples are added to specified wells in an uncoated microplate. Saline is added to all wells and the plate is allowed to incubate until the blood has eluted from the paper. Immediately following a transfer of a portion of this eluate into the antibodycoated microplate a horseradish peroxidase enzyme conjugated to 17-OHP (17-OHP: HRP) is added to the wells. A competition begins between the endogenous 17-OHP from the unknown samples, standards and controls and the 17-OHP: HRP for the limited number of antibody binding sites in the wells. An inverse relationship develops between the concentration of endogenous 17-0HP and the amount of 17-OHP: HRP, which will bind to the coated microplate. After a period of incubation the excess 17-OHP: HRP is removed by washing. A color developer, containing a colorless 3, 3', 5, 5'tetramethylbenzidine (TMB) and peroxide (H2O2), is added to all wells. The peroxidase react and subsequently the TMB is converted from color. The color development is terminated through the addition of a stopping reagent and the absorbance is measured at 650 nm using a microplate absorbance reader. A standard curve is constructed using the known concentration of each standard plotted against the corresponding absorbance reading of that standard. The concentrations of unknown controls and samples are determined by comparison to this standard curve.

An alternate procedure is described where a 1/8 inch disc from each blood spot is placed directly into designated wells of the coated microplate. The enzyme conjugated 17-OHP is added and the competition proceeds concurrent with the elution step. Upon completion period, the excess 17-OHP:HRP and the discs are removed by decanting, or aspirating the discs from each well. The plate is then washed and processed using the same procedure as described above.

Kit Contents

Materials Supplied

	Quantity per Kit
Wells	96	384	1920
Catalog No.	601501	601502	601503
Coated Microplates(Rabbit Anti-17-OHP)	1 Strip plate	4 plates	20 plates (5 bagsof 4 plates each)
Enzyme Conjugate Concentrate(17-OHP:HRP)*	2 mls	5 mls	25 mls
Conjugate Diluent*	20 mls	50 mls	250 mls

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Wash Buffer Concentrate (20X)	25 mls	50 mls	250 mls
Color Developer *	20 mls	55 mls	250 mls
Stopping Reagent	20 mls	55 mls	250 mls
Multi-Analyte Standards andControlsTSH/T4/17-OHP	1 set	1 set	4 sets

*These reagents are light sensitive. Avoid prolonged exposure to light.

NOTE: Do not use reagents or solutions that have become cloudy or discolored as these conditions indicate deterioration. The reagents should be stored in their original containers.

Do not exchange reagents from one kit with those of another kit unless the lot number and expiration of the reagent are the same.

Reagent Description

Coated Microplates (Rabbit Anti-17-OHP)

Microplates are coated with an antiserum produced in rabbits immunized with a 17-OHP hapten. The microplates are packaged in zipper-lock foil bags containing a desiccant. Store the unused microplates and/or strips from the strip plate in the zipper-lock bags with desiccant. Each microplate is labeled with a unique bar-code label.

Storage: Drv at 2-25° C

Expiration: Refer to the expiration date printed on the label

Enzyme Conjugate Concentrate (17-OHP: HRP)*

A 17-OHP derivative conjugated to horseradish peroxidase, danazol with an enzyme stabilizer.

*This reagent is light sensitive, plate store in the original brown confainer.

Enzyme Conjugate Concentrate must be diluted with Conjugate Diluent before use.

See Reagent Preparation Instructions section on page 11.

Storage: 2-8° C Protected From Light.

Expiration: Refer to the expiration date printed on the label.

Coniugate Diluent*

Tris-buffer, bovine serum and preservatives. * This reagent is light sensitive, store in the original brown container. Conjugate Diluent is used to dilute the conjugate concentrate before use. See Reagent Preparation Instructions section on page 11.

Storage: 2-8° C Protected from Light.

Expiration: Refer to the expiration date printed on the label.

Wash Buffer Concentrate (20X)

A concentrated solution of phosphate buffered saline containing a surfactant.

Wash Buffer Concentrate must be diluted with deionized or distilled water before use.

See Reagent Preparation Instructions section on page 11.

2-25° C Storage:

Expiration: Concentrate: Refer to the expiration date printed on the label.

Diluted wash buffer is stable for 1 month when stored at 2-25° C.

Color Developer*

Bottle contains a colorless solution of 3, 3', 5'-Tetramethylbenzidine in a diluted organic solvent with citrate buffer and hydrogen peroxide. *This reagent is light sensitive, store in the original brown container. This reagent should remain colorless; if it has discolored, discard it.

Storage: 2-8° C. Protected from light.

Expiration: Refer to the expiration date printed on the label.

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Stopping Reagent

Bottle contains a dilute solution of sodium fluoride (NaF) and a red dye. Storage: 2-8° C. Expiration: Refer to the expiration date printed on the label.

Multi-Analyte Standards and Controls

TSH, T4, 17-OHP

Multi-Analyte standards have been prepared from human whole blood, adjusted to a hematocrit of 55%. Each card of standards contains two circles each of six concentrations of 17-OHP at approximately zero. 10. 25. 50. 100 and 250 ng/ml serum equivalents. Control cards contain four circles each of three different concentrations of 17-OHP at approximately 15, 40, and 90 ng/ml serum equivalent. The standards and controls are spotted onto Whatman (previously known as Schleicher and Schuell (S&S®) 903") specimen collection paper. Refer to the information on the labels for the exact concentrations of the standards and acceptable ranges for the controls. Each set of standards and controls consists of two standard cards and one control card.

Drv at 2-8° C or below. Storage: Expiration: Refer to the expiration date printed on the label or Certificate of Analysis received with the kit.

Standard and control units are expressed in ng/ml. The values may be converted using the following formula.

1 nmol/l blood= 0.33 ng/ml blood

1 nmol/l blood= 0.73 ng/ml serum (at 55% hematocrit)

	Conversion Table
	Concentration in	Concentration in
	Serum ng/ml	Blood nmol/l
Standard Zero	0	0
Standard A	10	13.7
Standard B	25	34.2
Standard C	50	68.5
Standard D	100	137
Standard E	250	342
Control 1	15	20.6
Control 2	40	54.8
Control 3	90	123

This is an example only. Consult standard and control label or Certificate of Analysis received with the kit for exact concentrations.

Materials Required But Not Supplied for Both the Elution and Direct Procedures

1/8 inch (3 mm) diameter hole punch 1.
Forceps or fine tweezers to pick up the punched sample discs 2.
1. Pipettes to accurately dispense 100 ul volumes
1. Multi-channel pipettes to dispense 300 ul volumes or an automated plate washer
1. Microplate reader capable of reading at a wavelength of 650 nm
Plate rotator capable of 100-120 RPM or plate shaker. ి.
1. Microplate covers; plate sealers or low evaporation solid plastic microplate lids (e.g. NUNC lids with condensation rings and evaporation barriers)
1. Graduated cylinders
Deionized or distilled water တဲ

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Additional Materials Required for the Elution Procedure Only

Un-coated round bottom 96 well microplates

1. Saline (0.85% NaCl).
1. Multi-channel pipet with disposable tips to accurately transfer 15 µl of eluate.

Additional Materials Required for the Direct Procedure Only

Aspiration device capable of removing discs from coated plates.

1. Intended Use:
  The Accuwell" 17α-Hydroxyprogesterone EIA Kit is designed for the quantitative measurement of 17cc-Hydroxyprogesterone (17-OHP) concentrations in neonatal blood samples that have been collected onto Whatman [previously known as Schleicher and Schuell (S&S®) 903"] specimen collection paper. The results are used to screen newborns for classical congenital adrenal hyperplasia (CAH).

Characteristic:	Predicate Device:	Proposed Device:
	K 042425; Model B048-112Wallac Oy AutoDelfia Neonatal17α-HydroxyprogesteroneTime-resolvedflouoroimmunoassay	Neo-Genesis Accuwell17α-Hydroxyprogesterone EIA
Intended User	Clinical Laboratory Professionals	Clinical Laboratory Professionals
Intended Use	Measure 17α-Hydroxyprogesterone (17-OHP) levels in neonatal driedblood spots	Measure 17α-Hydroxyprogesterone (17-OHP) levels in neonatal driedblood spots
Indications for Use	Screening for increased levels of17-OHP in newborns	Screening for increased levels of17-OHP in newborns
Chemical Principle	Competitive reaction betweeneuropium-labeled 17-OHPand sample 17-OHP for alimited number of bindingsites on 17-OHP specificpolyclonal antibodies derivedfrom rabbit	Competitive reaction betweenhorseradish peroxidase(HRP) labeled 17-OHP andsample 17-OHP for a limitednumber of binding sites on17-OHP specific polyclonalantibody derived from rabbit
Assay Type	Time-resolvedfluoroimmunoassay	Enzyme-immunoassay
Detection Method	Europium ions dissociated fromthe labeled antiserum formhighly fluorescent chelateswith components of anenhancement solution.Fluorescence in each well isthen measured	Peroxidase which remains boundto micro-wells reacts withperoxide and TMBsubsequently converting theTMB from colorless to a bluecolor which is measured
Assay Processing Method	Automated	Manual
Detection Equipment	Autodelfia - time resolvedfluorometer	Microplate Spectrophotometer
Sample Requirements	Newborn blood collected onS&S 903 filter paper orequivalent collection cards	Newborn blood collected onS&S 903 filter paper

1. Summary of Technological Characteristics 1,2:

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. .

Specimen Rejection Criteria	Sample spot not uniformlysaturated with blood; samplespots punched to close to theedge of the blood spot;poorly collected andimproperly dried specimens;non-eluting blood spot;contamination of blood spotfilter paper with foreignmaterial		Sample spot not uniformlysaturated with blood; samplespots punched to close to theedge of the blood spot;poorly collected andimproperly dried specimens
Specimen	1/8 inch spot (3.2mm) punchedfrom a standard collectioncard		1/8 inch spot (3.2mm) punchedfrom a standard collectioncard of S&S 903 paper
Standard Configuration	Human blood with a hematocritof 50-55% and spotted onS&S 903 filter paper.Calibrated using gravimetricmethods. Seven levels		Human whole blood adjusted to55% hematocrit and spottedon S&S 903 filter paper.Calibrated using gravimetricmethods. Six levels
Standard Range	0 - 190 ng/ml approximateserum equivalent		0 - 250 ng/ml approximateserum equivalent
Control Configuration	Human blood with a hematocritof 50-55% and spotted onS&S 903 filter paper.Calibrated using gravimetricmethods. Two levels		Human whole blood adjusted to55% hematocrit and spottedon S&S 903 filter paper.Calibrated using gravimetricmethods. Three levels
Control Levels	14 and 40 ng/ml approximateserum equivalent		15, 40 and 90 ng/ml approximateserum equivalent
Calculation of Results-Recommendations	Spline Smoothed regressionanalysis is used to calibrateeach plate with the LOG ofthe concentration on the X-axis and B/Bmax on the Y-axis (response).		Five parameter logistic plot ofabsorbance versusconcentration
Analytical Sensitivity	1.5 ng/ml		2.2 ng/ml
Within Run Precision 3,4	Mean (ng/ml)% cvMean (ng/ml)% cvMean (ng/ml)% CV	8.811.329.38.962.18.9	Mean (ng/ml)% CVMean (ng/ml)% CVMean (ng/ml)% CV	28.23.556.13.7108.12.7
Between Run Precision 3,4	Mean (ng/ml)% CVMean (ng/ml)% CVMean (ng/ml)% CV	8.84.029.33.362.12.4	Mean (ng/ml)% CVMean (ng/ml)% CVMean (ng/ml)% CV	28.210.356.28.0108.111.2
Total Precision 3,4	Mean (ng/ml)% CVMean (ng/ml)% CVMean (ng/ml)% CV	8.812.029.39.562.19.2	Mean (ng/ml)% CVMean (ng/ml)% CVMean (ng/ml)% CV	28.111.056.18.9108.111.6

11 - 11

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...

Interfering SubstancesCross Reacting SubstancesGreater than 0.01 %:	EDTA or citrate blood	Lipemia
Progesterone	3.60 %	0.5 %
21-Desoxycortisol	1.75 %	2.42 %
16a-Hydroxyprogesterone	0.3 %	1.2 %
11-Desoxycortisol	0.4 %	0.6 %
11-Deoxy-17-hydroxycorticosterone	0.33 %	< 0.01 %
17a-Hydroxypregnenolone	0.13 %	< 0.01 %
Deoxicorticosterone	0.3 %	0.06 %
5-Pregnen-3β,17-Diol-20-one 3sulfate	< 0.01 %	0.2 %
Cortisol	< 0.01 %	0.06 %
Tetrahydrocortisone	0.02 %	< 0.01 %
Expected Values-Term (normal birth weight)	3-5 days old, >2000g birthweight(unknown percentile):	1-3 days old, ≥ 2500g birthweight(99th percentile):
	normal < 22 ng/ml	ON Direct:normal < 27.8 ng/ml
	request 2nd sample 22-44 ng/ml	Follow-up ≥ 27.8 ng/ml
	inform physician > 44 ng/ml	ON Eluate:normal < 23.9 ng/ml
		Follow-up ≥ 23.9 ng/ml
		3Hr Eluate:normal < 24.3 ng/ml
		Follow-up ≥ 24.3 ng/ml
		> 4 days old, ≥ 2500g birthweight(99th percentile):
		ON Direct:normal < 21.1 ng/ml
		Follow-up ≥ 21.1 ng/ml
		ON Eluate:normal < 19.5 ng/ml
		Follow-up ≥ 19.5ng/ml
		3Hr Eluate:normal < 19.4 ng/ml
		Follow-up ≥ 19.4 ng/ml
Expected Values-Premature (low birth weight)	3-5 days old, <2000g birthweight(unknown percentile):	1-3 days old, < 2500g birthweight(99th percentile):
	normal < 44 ng/ml	ON Direct:normal < 46.4 ng/ml
	request 2nd sample 44-73 ng/ml	Follow-up ≥ 46.4 ng/ml
	inform physician > 73 ng/ml	ON Eluate:normal < 40.9 ng/ml
		Follow-up ≥ 40.9 ng/ml
		3Hr Eluate:normal < 40.5 ng/ml
		Follow-up ≥ 40.5 ng/ml

: -

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. .

Method Comparison
	The AutoDelfia Neonatal17-OHP kit was compared withthe DELFIA Neonatal 17-OHP kit using 1579 infantblood spot specimens.Results showed a meanvalue of 13.3 ng/ml serumfor the AutoDelfia and 12.6ng/ml serum for thepredicate DELFIA kit.	The Accuwell 17-OHP wascompared to the Autodelfia 17-OHP kit using whole blood spotspecimens from the followinginfant populations:>2500g b.w., 0-1 days old (n=133)-AutoDelfia Mean (ng/ml) =22.6Accuwell Means (ng/ml):ON Direct = 16.5ON Eluate = 16.73Hr Eluate = 17.3>2500g b.w., 2-3 days old(n=133)-AutoDelfia Mean (ng/ml) =16.2Accuwell Means (ng/ml):ON Direct = 11.3ON Eluate = 12.23Hr Eluate = 12.8>2500g b.w., >4 days old (n=197)-AutoDelfia Mean (ng/ml) =9.5Accuwell Means (ng/ml):ON Direct = 7.3ON Eluate = 7.83Hr Eluate = 8.31400-2500g b.w., 1-6 days (n=30)-AutoDelfia Mean (ng/ml) =25.5Accuwell Means (ng/ml):ON Direct = 19.3ON Eluate = 19.23Hr Eluate = 19.3

.. .

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CDC Control Results
CDC Lot 451 - Enriched with25 ng/ml serum	Mean (ng/ml)% cv	30.011.3	Mean (ng/ml)% cv	28.111.0
CDC Lot 452 - Enriched with50 ng/ml serum	Mean (ng/ml)% cv	59.310.6	Mean (ng/ml)% cv	56.18.9
CDC Lot 453 - Enriched with100 ng/ml serum	Mean (ng/ml)% cv	120.210.1	Mean (ng/ml)% cv	108.111.6
Intercept =Slope =		- 0.451.2049		+ 2.11.0629

1 Data displayed for the predicate device derived from the directions for use unless otherwise noted.

2 Data displayed for the Accuwell 17-OHP kit was derived using only the overnight eluate method unless otherwise noted.

Results from the predicate device were obtained from their published directions. The method used or definitions were not 3 declared, therefore they may or may not use the same calculation method.
4 Accuwell 17-OHP precision studies were conducted in accordance with NCCLS EP5-A2, "Evaluation of Precision Performance of Quantitative Measurement Methods"; Approved Guideline-Second Edition. All precision testing was r of orned by Neo-Genesis at Neo-Genesis' Portland, OR manufacturing facility. Sources of variability included the use of different; operators, days and reagent lot numbers.
5 Results from an in house study of randomly selected samples from newborns with birth weight >2200 grams. These results are presented as an example only and used strictly as guidance and should not be used to establish reference ranges.
6 Results from an in house study of randomly selected samples from newborns with birth weight ≤2200 grams. These results roodito for an in housed strictly as guidance and should not be used to establish reference ranges.
Predicate kit results for 17-OHP were derived from CDC publication "Newborn Screening Quality Assurance Program 2005 7 Annual Summary Report, Vol. 23, Jan 2006. % CV was calculated using the reported mean and the average within laboratory standard deviation. Intercept and slope are as reported.
1. Clinical and Non-Clinical Data:

Precision

Precision studies were conducted in accordance with NCCLS EP5-A2, "Evaluation of Precision Performance of Quantitative Measurement Methods"; Approved Guideline-Second Edition.

Testing included one run per day, with two sample aliquots per run, performed on 20 different days using each of three different assay procedures. The three procedures were designated as: 1) "ON Eluate", 2) "3 Hr Eluate" and 3) "ON Direct DBS", respectively. The resulting data were used to estimate repeatability, between-day and within-device precision as described by the standard, for each procedure.

All precision testing was performed by Neo-Genesis' Portland, OR manufacturing facility. Sources of variability included the use of different: operators, days and reagent lot numbers.

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

	Sample ID:
		CDC 451*(25+ ng/ml)	CDC 452(50+ ng/ml)	CDC 453(100+ ng/ml)
	Count	40	40	40
	Mean	28.2	56.1	108.1
ON Eluate	SD	3.0	4.9	12.3
	%CV	10.7	8.8	11.4
Within-Run Standard Deviation:	Sr	1.0	2.1	2.9
Between-Day Standard Deviation:	Sdd	2.9	4.5	12.1
Within Device Standard Deviation:	ST	3.1	5.0	12.5
	Count	40	40	40
	Mean	26.2	53.0	104.6
3 HR Eluate	SD	3.4	4.1	11.9
	%CV	12.9	7.7	11.3
Within-Run Standard Deviation:	Sr	1.9	2.2	5.4
Between-Day Standard Deviation:	Sdd	2.9	3.4	10.7
Within Device Standard Deviation:	ST	3.4	4.1	12.0
ON Direct DBS	Count	40	40	40
	Mean	29.5	57.9	108.6
	SD	4.2	5.8	13.2
	%CV	14.1	10.0	12.2
Within-Run Standard Deviation:	Sr	1.5	4.4	11.7
Between-Day Standard Deviation:	Sdd	3.9	3.9	6.2
Within Device Standard Deviation:	ST	4.2	5.8	13.3

3 ummary Results and Estimates of Precision for Accuwell 17-OHP EIA Kit
-------------------------------------------------------------------------

*Enriched as reported by Centers for Disease Control and Prevention. Newborn Screening Quality Assurance Program Annual Report 2005

Analytical Sensitivity

The analytical sensitivity was determined for each procedure including Overnight Eluate, 3 hour Eluate, and the Overnight Direct. Sensitivity was defined by the calculated concentration that corresponds to the mean of the absorbance of the zero standard (N=26) minus two times the standard deviation of those same absorbance measurements.

The zero standard was tested multiple time (N=26) in one assay. Testing was performed in each of the two procedures, eluate and direct, and spanning the range of allowable elution times, three hours and overnight for the eluate procedure.

Extrapolation beyond the upper and lower limits of the standard curve or beyond the limits of detection is not an acceptable laboratory practice. Therefore any sample result falling below these calculated concentrations should be reported as less than the limit of detection.

This data is provided for example only and should be confirmed by each laboratory and appropriate concentrations defined.

	Overnight Eluate	3 Hour Eluate	Overnight Direct
Absorbance Mean	2.506	1.648	2.604
Standard Deviation	0.04	0.02	0.06
Absorbance -2 SD	2.426	1.608	2.484
Calculated Concentration (ng/ml)	2.2	1.5	2.4

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

Linearity and Recovery

The Accuwell 17-OHP Kit linear range and the measuring (reportable) range is approximately 2.0 ng/ml to 250 ng/ml based on study data described below, the sensitivity study described elsewhere and limited by the value assigned to the highest standard of the calibration curve.

Assay results obtained outside the measuring (reportable) range should be reported as either "less than" or "greater than" the established low or high limit, respectively, as applicable to the individual result.

Dried blood spots representing a range of sample concentrations were prepared for the study. The sample analyte levels in ng/ml were: 10, 24.5, 39, 68, 97, 126, 155, 184, 213, 242, 271 and 300 (n = 12 concentrations).

Four aliquots of each sample of the range of samples prepared, were tested in each of two runs using each of three different assay procedures. The three procedures were designated as:1) "ON Eluate", 2) "3 Hr Eluate" and 3) "ON Direct DBS", respectively. The mean of the total number of values obtained for each sample (n=8) within each procedure, was compared to the samples expected value. Results of the comparisons are described below.

			ON Eluate		3 Hr Eluate	ON Direct DBS
Linear Regression Analysis			$y = 1.0036x - 0.3475$$R^2 = 0.9994$			$y = 1.1236x - 9.3625$$R^2 = 0.992$		$y = 1.1362x - 9.5154$$R^2 = 0.99$
Sample #	Expected(ng/ml)	Actual(ng/ml)	Recovery(%)	Actual(ng/ml)	Recovery(%)	Actual(ng/ml)	Recovery(%)
1	300	298.1	99.4	354.4	118.1	319.9	106.6
2	271	274.4	101.3	284.4	104.9	324.1	119.6
3	242	241.9	100.0	257.9	106.6	261.6	108.1
4	213	212.3	99.7	221.7	104.1	244.0	114.6
5	184	182.9	99.4	194.1	105.5	185.9	101.0
6	155	157.2	101.4	159.8	103.1	157.5	101.6
7	126	131.7	104.5	127.7	101.3	128.5	102.0
8	97	95.2	98.1	94.2	97.1	92.6	95.5
9	68	66.2	97.4	66.4	97.6	69.9	102.8
10	39	38.2	97.9	37.0	94.9	33.3	85.4
11	24.5	23.0	93.9	23.0	93.9	22.8	93.1
12	10	10.4	104.0	10.3	103.0	10.7	107.0
Average Recovery-Overall			99.7		102.5		103.1
Minimum			93.9		93.9		85.4
Maximum			104.5		118.1		119.6

Results of Assay Linearity and % Recovery Study for Accuwell 17-OHP EIA Kit

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

Specificity

Cross Reactivity

The cross reactivity percentage is determined by dividing the concentration of analyte (17-OHP) at 50% displacement by the concentration of the interferant at 50% displacement from the absorbance corresponding to the zero standard.

Trivial Name	CrossReactivity	Trivial Name	CrossReactivity
21-Desoxycortisol	2.4 %	Cholesterol	< 0.01 %
16α-Hydroxyprogestrone	1.2 %	Corticosterone	< 0.01 %
11-Desoxycortisol	0.6%	Cortisol Glucuronide	< 0.01 %
Progesterone	0.5 %	Cortisone	< 0.01 %
5-Pregnen-3β,17-Diol-20-one 3sulfate	0.2 %	Cortisone 21-Sulphate Sodium Salt	< 0.01 %
Cortisol	0.06 %	Dehydroisoandrosterone	< 0.01 %
Desoxycorticosterone	0.06 %	Dehydroepiandrosterone SodiumSulphate	< 0.01 %
1-Dehydrotestosterone	< 0.01 %	Dexamethasone	< 0.01 %
5β-Dihydrocortisol	< 0.01 %	Estriol	< 0.01 %
5β-Dihydrocortisone	< 0.01 %	Estrone	< 0.01 %
6β-Hydroxycortisol	< 0.01 %	Prednisolone	< 0.01 %
11-Dehydrocorticosterone	< 0.01 %	Prednisone	< 0.01 %
16α-Hydroxypregnenolone	< 0.01 %	Pregnenolone	< 0.01 %
17α-Estradiol	< 0.01 %	Pregnenolone Sulphate, Sodium Salt	< 0.01 %
17α-Hydroxypregnanolone	< 0.01 %	Spironolactone	< 0.01 %
17β-Estradiol	< 0.01 %	Testosterone	< 0.01 %
20α-Hydroxyprogesterone	< 0.01 %	Tetrahydrocortisol	< 0.01 %
Aldosterone	< 0.01 %	Tetrahydrocortisone	< 0.01 %

Interfering Substances

Interferences due to hemoglobin, conjugated and unconjugated bilirubin, and lipids were studied using methods described in NCCLS (document EP7-A.) as guidance. 22 Hemoglobin and bilirubin (conjugated and unconjugated) caused no detectable interference. Significant interference was seen from the lipid solutions added to the Accuwell 17-OHP test.

Method Comparison

A retrospective study was conducted to compare the results obtained with the Accuwell 17-OHP EIA to those obtained by a currently marketed neonatal 17-OHP screening device. Test samples were submitted to the study as blinded neonatal dried blood spots collected in sequence under routine screening conditions from a U.S. department of public health laboratory. Original screening results for each sample using the predicate test kit were also obtained from the submitting laboratory for method comparison.

A summary of results of the method comparisons are provided in Tables 9 -- 20, below. (Samples interpreted below as "Follow-Up" were not confirmed.)

{12}------------------------------------------------

Table 9: Predicate vs Accuwell O/N Direct

Values at the 99th, 97.5", and 95th Percentiles for a Population of Babies >2500 gm and 0-1 Days of Age at Sample Collection

99th Percentile			97.5th Percentile			95th Percentile
	Accuwell Normal	Follow-up		Accuwell Normal	Follow-up		Accuwell Normal	Follow-up
Predicate	< 27.5	≥ 27.5 Totals:	Predicate	< 25.8	≥ 25.8 Totals:	Predicate	< 21.8	≥ 21.8 Totals
Normal	< 43.8N=130	114 16 130	Normal	< 43.8N=130	112 18 130	Normal	< 43.8N=130	100 30 130
Follow-Up	≥ 43.8N = 3	0 3 3	Follow-Up	≥ 43.8N = 3	0 3 3	Follow-Up	≥ 43.8N = 3	0 3 3
Totals:	114	19 133	Totals:	112	21 133	Totals:	100	33 133

The Accuwell 17-OHP EIA (ON Direct) results for this population were: mean =16.5 ng/ml, with a range of 3.7 to 85.6 ng/ml; while the predicate kit yielded: mean =22.6 ng/ml, with a range of 10.1 to 86.0 ng/ml.

The correlation was found to be: y (Predicate) = 0.970 (Accuwell ON Direct) + 6.569, R = 0.9570

Table 10: Predicate vs Accuwell O/N Eluate

Values at the 99th, 97.5th, and 95th, Percentiles for a Population of Babies ≥2500 gm and 0-1 Days of Age at Sample Collection

99th Percentile				97.5th Percentile				95th Percentile
	Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up
Predicate		< 23.9	≥ 23.9 Totals:	Predicate		< 22.3	≥ 22.3 Totals:	Predicate		< 20.2	≥ 20.2 Totals
Normal	< 43.8N=130	106	24 130	Normal	< 43.8N=130	103	27 130	Normal	< 43.8N=130	101	29 130
Follow-Up	≥ 43.8N = 3	0	3 3	Follow-Up	≥ 43.8N=3	0	3 3	Follow-Up	≥ 43.8N = 3	0	3 3
Totals:		106	27 133	Totals:		103	30 133	Totals:		101	32 133

The Accuwell 17-OHP ElA (ON Eluate) results for this population were: mean =16.7 ng/ml, with a range of 5.3 to 65.6 ng/ml; while the predicate kit yielded: mean =22.6 ng/ml, with a range of 10.1 to 86.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.049 (Accuwell ON Direct) + 5.066, R = 0.9257

Table 11: Predicate vs Accuwell 3 Hour Eluate

Values at the 99th, 97.5th, and 95th Percentiles for a Population of Babies ≥2500 gm and 0-1 Days of Age at Sample Collection

99th Percentile				97.5th Percentile				95th Percentile
	Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up
Predicate		< 23.0	≥ 23.0	Predicate		< 21.7	≥ 21.7	Predicate		< 19.8	≥ 19.8
Normal	< 43.8N=130	104	26	Normal	< 43.8N=130	103	27	Normal	< 43.8N=130	99	31
Follow-Up	≥ 43.8N = 3	0	3	Follow-Up	≥ 43.8N = 3	0	3	Follow-Up	≥ 43.8N = 3	0	3
Totals:		104	29	Totals:		103	30	Totals:		99	34

The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean =17.3 ng/ml, with a range of 5.1 to 71.1 ng/ml; while the predicate kit yielded: mean =22.6 ng/ml, with a range of 10.1 to 86.0 ng/ml..

The correlation was found to be: y (Predicate) = 0.998 (Accuwell ON Direct) + 5.410, R = 0.9291

{13}------------------------------------------------

Table 12: Predicate vs Accuwell O/N Direct

Values at the 95th Percentile for a

Population of Babies >2500 gm and 2-3 Days of Age at Sample Collection

95th Percentile
	Accuwell Normal	Follow-up
Predicate
	< 18.5	≥ 18.5
Normal	< 29.2N=118
	113	5
Follow-Up	≥ 29.2N = 15
	0	15
	Totals:
	113	20
	Totals:
	118	15
		133

The Accuwell 17-OHP EIA (ON Direct) results for this population were: mean =11.3 ng/ml, with a range of 3.4 to 81.1 ng/ml; while the predicate kit yielded: mean =16.2 ng/ml, with a range of 6.9 to 81.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.045 (Accuwell ON Direct) + 4.392, R = 0.9785

Table 13: Predicate vs Accuwell O/N Eluate Values at the 95th Percentile for a

Population of Babies ≥2500 gm and 2-3 Days of Age at Sample Collection

95th Percentile
	AccuwellNormal	Follow-up
Predicate	< 18.7	≥ 18.7	Totals:
Normal	< 29.2N=118	114	4	118
Follow-Up	≥ 29.2N = 15	0	15	15
	Totals:	114	19	133

The Accuwell 17-OHP EIA (ON Eluate) results for this population were: mean =12.2 ng/ml, with a range of 3.4 to 67.5 ng/ml; while the predicate kit yielded: mean =16.2 ng/ml; with a range of 6.9 to 81.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.156 (Accuwell ON Eluate) + 2.030, R = 0.9729

Table 14: Predicate vs Accuwell 3 Hour Eluate

Values at the 95t" Percentile for a

Population of Babies >2500 gm and 2-3 Days of Age at Sample Collection

95th Percentile
	Accuwell	Follow-up
Predicate	Normal
	< 17.7	≥ 17.7	Totals:
Normal	< 29.2N=118	115	3	118
Follow-Up	≥ 29.2N = 15	0	15	15
	Totals:	115	18	133

The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean =12.8 ng/ml, with a range of 4.5 to 69.6 ng/ml; while the predicate kit yielded: mean =16.2 ng/ml, with a range of 6.9 to 81.0 ng/ml.

The correlation was found to be: y (Predicate) = 1.150 (Accuwell ON Direct) + 1.495, R = 0.9691

{14}------------------------------------------------

Table 15: Predicate vs Accuwell O/N Direct
Values at the 97.5" and 95th Percentiles for a
Population of Babies ≥2500 gm and ≥ 4 Days of Age at Sample Collection

97.5th Percentile			95th Percentile
	Accuwell Normal	Follow-up		Accuwell Normal	Follow-up
Predicate	< 14.8 ≥ 14.8 Totals:	Predicate		< 11.5 ≥ 11.5 Totals:
Normal	< 21.9N=192	181 11 192	Normal	< 21.9N=192	170 22 192
Follow-Up	≥ 21.9N = 5	0 5 5	Follow-Up	≥ 21.9N = 5	0 5 5
	Totals:	181 16 197		Totals:	170 27 197

The Accuwell 17-OHP EIA (ON Direct) results for this population were: mean =7.3 ng/ml, with a range of 2.2 to 22.6 ng/ml; while the predicate kit yielded: mean =9.5 ng/ml, with a range of 2.7 to 31.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.090 (Accuwell ON Direct) + 1.565, R = 0.9538

Table 16: Predicate vs Accuwell O/N Eluate

Values at the 97.5" and 95th Percentiles for a Population of Babies >2500 gm and > 4 Days of Age at Sample Collection

97.5th Percentile			95th Percentile
	Accuwell Normal	Follow-up		Accuwell Normal	Follow-up
Predicate	< 15.2 $\ge$ 15.2 Totals:		Predicate	< 13.6 $\ge$ 13.6 Totals:
Normal	< 21.9N=192 181 11 192		Normal	< 21.9N=192 178 14 192
Follow-Up	$\ge$ 21.9N = 5 1 4 5		Follow-Up	$\ge$ 21.9N = 5 0 5 5
	Totals: 182 15 197			Totals: 178 19 197

The Accuwell 17-OHP EIA (ON Eluate) results for this population were: mean =7.8 ng/ml, with a range of 1.4 to 26.5 ng/ml; while the predicate kit yielded: mean =9.5 ng/ml, with a range of 2.7 to 31.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.070 (Accuwell ON Eluate) + 1.172, R = 0.9142

Population of Babies ≥2500 gm and ≥ 4 Days of Age at Sample Collection
97.5th Percentile		95th Percentile
Predicate	Accuwell Normal	Follow-up	Accuwell Normal	Follow-up
	< 15.6	≥ 15.6	< 14.1	≥ 14.1
Normal	< 21.9N=192176	16	< 21.9N=192174	18
Follow-Up	≥ 21.9N = 50	5	≥ 21.9N = 50	5
Totals:	176	21	174	23
Totals:	192		192
	197		197

Table 17: Predicate vs Accuwell 3 Hour Eluate Values at the 97.5th and 95th Percentiles for a

ﺍﻟﻤﻌﺎﻟﻢ Collecti t

The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean =8.3 ng/ml, with a range of 2.2 to 28.4 ng/ml; while the predicate kit yielded: mean =9.5 ng/ml, with a range of 2.7 to 31.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.024 (Accuwell 3Hr Eluate) + 1.081, R = 0.9350

{15}------------------------------------------------

Predicate vs Accuwell O/N Direct Table 18: Values at the 99th and 97.5th Percentiles for a Population of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection

99th (100th) Percentile				97.5th Percentile
	Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up
AutoDelFIA	< 46.0	≥ 46.0	Totals:	AutoDelFIA	< 43.9	≥ 43.9	Totals:
Normal	< 45.1 N=28	28	0	Normal	< 40.3 N=26	26	0
Follow-Up	≥ 45.1N = 2	1	1	Follow-Up	≥ 40.3N = 4	3	1
Totals:	29	1	30	Totals:	29	1	30

The Accuwell 17-OHP EIA (ON Direct) results for this population were: mean =19.3 ng/ml, with a range of 6.5 to 47.8 ng/ml; while the predicate kit yielded: mean =25.5 ng/ml, with a range of 9.0 to 65.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.22 (Accuwell ON Direct) + 1.971, R = 0.9556

Table 19: Predicate vs Accuwell O/N Eluate

Values at the 99th and 97.5th Percentiles for aPopulation of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection
99th (100th) Percentile				97.5th Percentile
Accuwell	Normal	Follow-up		Accuwell	Normal	Follow-up
AutoB-IFA	< 31.7	≥ 31.7	Totals:	AutoB-IFA	< 28.4	≥ 28.4	T

		< 31.7	≥ 31.7	Totals:			< 28.4	≥ 28.4	Totals
AutoDelFIA					AutoDelFIA
Normal	< 45.1N=28	24	4	28	Normal	< 40.3N=26	23	3	26
Follow-Up	≥ 45.1N = 2	1	1	2	Follow-Up	≥ 40.3N = 4	1	3	4
	Totals:	25	5	30		Totals:	24	6	30

The Accuwell 17-OHP EIA (ON Eluate) results for this population were: mean =19.2 ng/ml, with a range of 6.7 to 40.2 ng/ml; while the predicate kit yielded: mean =25.5 ng/ml, with a range of 9.0 to 65.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.234 (Accuwell ON Direct) + 1.812, R = 0.8770

Population of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection
99th (100th) Percentile			97.5th Percentile
	Accuwell	Normal	Follow-up	Accuwell	Normal	Follow-up
AutoDelFIA		< 33.3	≥ 33.3	AutoDelFIA		< 27.7	≥ 27.7
Normal	< 45.1N=28	26	2	Totals: 28	Normal	< 40.3N=26	23	3	Totals: 26
Follow-Up	≥ 45.1N = 2	1	1	Totals: 2	Follow-Up	≥ 40.3N = 4	2	2	Totals: 4
	Totals:	27	3	30		Totals:	25	5	30

Predicate vs Accuwell 3Hr Eluate Table 20: Values at the 99th and 97.5th Percentiles for a

nies 1400 - 2500 gm and 1-6 Davs of Age at S

at Ca

The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean =19.3 ng/ml, with a range of 6.6 to 39.2 ng/ml; while the predicate kit yielded: mean =25.5 ng/ml, with a range of 9.0 to 65.0 ng/ml..

The correlation was found to be: y (Predicate) = 1.174 (Accuwell 3Hr Eluate) + 2.795, R = 0.8421

{16}------------------------------------------------

Image /page/16/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three lines forming its body and wings. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Neo-Genesis c/o Jannet Baldwin President, Genesis Northwest, Inc. 15140 SE 82nd Drive Clackamas, OR 97015

MAR 1 3 2007

Re: K060452

Trade Name: Accuwell 17-a-Hydroxyprogesterone Neonatal Screening Enzyme Immunoassay Regulation Number: 21 CFR 862.1395 Regulation Name: 17-Hydroxyprogesterone test system. Regulatory Class: Class I Product Code: JLX Dated: January 31, 2007 Received: February 2, 2007

Dear Ms. Baldwin:

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 either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

{17}------------------------------------------------

Page 2 -

This letter will allow you to begin marketing your device as described in your Section 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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0490. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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 (240) 276-3150 or at its Internet address at http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours.

Jean M. Cooper, M.S., D.V.M.

Jean M. Cooper, M.S., D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

{18}------------------------------------------------

Indications for Use

510(k) Number (if known):

K060452

Device Name:

Accuwell 17 a-Hydroxyprogesterone Neonatal Screening Enzyme Immunoassay

Indications For Use:

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (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)

Carol Benson

office of In Vitro Diagnostic Device valuation and Safety

Page 1 of

K 060452

Regulation Number and Section

§ 862.1395 17-Hydroxyprogesterone test system.

(a)
Identification. A 17-hydroxyprogesterone test system is a device intended to measure 17-hydroxyprogesterone (a steroid) in plasma and serum. Measurements of 17-hydroxyprogesterone are used in the diagnosis and treatment of various disorders of the adrenal glands or the ovaries.(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 § 862.9.