ACCUWELL 17-ALPHA-HYDROXYPROESTERONE ENZYME IMMUNOASSAY WITH MODELS 6015XX-ECAH

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k060452 B. Purpose for Submission: New Device C. Measurand: 17-alpha-Hydroxyprogesterone D. Type of Test: Quantitative, Enzyme Immunoassay E. Applicant: Neo-Genesis F. Proprietary and Established Names: Accuwell 17-Alpha-Hyderoxyprogesterone Enzyme Immunoassay with Models 6015XX-ECAH G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | JLX- Radioimmunoassay, 17-Hydroxyprogesterone | Class I, meets the limitation of exemptions, 21 CFR 862.9 (c), (2) | 21 CFR 862.1395, 17-Hydroxy-progesterone test system. 21 CFR 862.9 (c), (2) | 75 Clinical chemistry (CH) | H. Intended Use: 1. Intended use(s): {1} See indications for use below. 2. Indication(s) for use: 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). 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: Microplate reader capable of reading a wavelength of 650 nm I. Device Description: The Accuwell 17α-Hydroxyprogesterone EIA Kit contains the following: - Microplates coated with rabbit anti-17-OHP. - Enzyme conjugate which contains 17-OHP derivative conjugated to horseradish peroxidase. - Conjugate diluent which contains tris-buffer, bovine serum and preservatives. - Wash buffer concentrate which is a concentrated solution of phosphate buffered saline containing a surfactant. - Color developer which contains 3,3',5,5'-Tetramethylbenzidine in a diluted organic solvent with citrate buffer and hydrogen peroxide. - Stopping reagent which is a dilute solution of sodium fluoride and a red dye. - Multi-Analyte standards (0, 10, 25, 50, 100 and 250 ng/mL) serum equivalents. Each card of standards is made from human whole blood, adjusted to a hematocrit of 55% and each level is then spotted onto Whatman 903 specimen collection paper. - Controls (15, 40 and 90 ng/mL) serum equivalent. Each control card is made from human whole blood, adjusted to a hematocrit of 55% and each level is then spotted onto Whatman specimen collection paper. Human source material was tested and found negative for HIV 1 and 2, HBV and HCV using FDA approved methods. J. Substantial Equivalence Information: 1. Predicate device name(s): {2} Wallac Oy AutoDelfia Neonatal 17α-Hydroxyprogesterone Time-resolved fluoroimmunoassay 2. Predicate 510(k) number(s): k042425 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | Device | Predicate | | Indications for use | Screening for increased levels of 17-OHP in newborns | Screening for increased levels of 17-OHP in newborns | | Sample Requirements | Neonatal blood collected on Whatman 903 filter paper | Neonatal blood collected on Whatman 903 filter paper | | Specimen | 1/8 inch spot punched from a standard collection card of Whatman 903 paper | 1/8 inch spot punched from a standard collection card of Whatman 903 paper | | Differences | | | | --- | --- | --- | | Item | Device | Predicate | | Assay Type | Enzyme-immunoassay | Time-resolved fluoroimmunoassay | | Detection Method | Peroxidase which remains bound to micro-wells reacts with peroxide and TMB subsequently converting the TMB from colorless to a blue color which is measured | Europium ions dissociated from the labeled antiserum form highly fluorescent chelates with components of an enhancement solution. Fluorescence in each well is then measured | | Standard Range | 3.8 to250 ng/mL approximate serum equivalent | Up to 190 ng/mL approximate serum equivalent | | Controls | Two levels concentration 14 and 40 ng/mL | Three levels concentration 15, 40 and 90 ng/mL | K. Standard/Guidance Document Referenced (if applicable): {3} 4 | Area of Study | Reference Procedure | Reference Title | | --- | --- | --- | | Stability Testing | CEN | Stability of In Vitro Diagnostic reagents (13640) | | Interference | CLSI/NCCLS EP7-A | Interference Testing in Clinical Chemistry | | Method Comparison | CLSI/NCCLS EP9-A | Method Comparison and Bias Estimation using Patient Samples | | Precision | CLSI/NCCLS EP5-A | Evaluation of Precision Performance of Quantitative Measurement Methods | L. Test Principle: The samples can be processed with two different methods; (1) a 1/8 inch disc from each blood spot is placed directly into designated wells of the coated microplate with an overnight incubation, (2) An elution is done using an 1/8 inch disc from each blood spot which is placed in the designated well of an uncoated microplate and saline is added to elute the blood from the dried blood spot. A portion of the eluate is then transferred into a designated well of the coated microplate with an overnight incubation or for 3 hours. The Accuwell 17α-hydroxyprogesterone immunoassay (EIA) is based on the competition principle. An unknown amount of antigen present in the sample and a fixed amount of enzyme labeled antigen compete for the binding sites of the antibodies coated onto the wells. After incubation the wells are washed to stop the competition reaction and the color developer is added. The color development is terminated by the addition of the stopping reagent and the absorbance measured at 650 nm and is inversely proportional to the amount of the antigen in the sample. Results of samples can be determined directly using the standard curve. M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: Precision was assessed using three neonatal dried bloodspots prepared by the Centers for Disease Control and Prevention Neonatal Screening Quality Control Program. Samples were prepared using three different assay procedures: Over night Eluate (ON Eluate), Three hour Eluate (3 Hr Eluate) and Over Night Direct (ON Direct). Each of these samples was assayed in duplicate once a day for 20 days. Results are presented in the table below: | | ON Eluate | | | 3 Hr Eluate | | | ON Direct | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Sample ID | CDC 451 | CDC 452 | CDC 453 | CDC 451 | CDC 452 | CDC 453 | CDC 451 | CDC 452 | CDC 453 | {4} | Number | 40 | 40 | 40 | 40 | 40 | 40 | 40 | 40 | 40 | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Mean | 28.2 | 56.1 | 108.1 | 26.2 | 53.0 | 104.6 | 29.5 | 57.9 | 108.6 | | SD | 3.0 | 4.9 | 12.3 | 3.4 | 4.1 | 11.9 | 4.2 | 5.8 | 13.2 | | %CV | 10.7 | 8.8 | 11.4 | 12.9 | 7.7 | 11.3 | 14.1 | 10.0 | 12.2 | | Within-Run SD | Sr | 1.0 | 2.1 | 2.9 | 1.9 | 2.2 | 5.4 | 1.5 | 4.4 | | Between-Day SD | Sdd | 2.9 | 4.5 | 12.1 | 2.9 | 3.4 | 10.7 | 3.9 | 3.9 | | Within Device SD | ST | 3.1 | 5.0 | 12.5 | 3.4 | 4.1 | 12.0 | 4.2 | 5.8 | | Daily Mean Standard Error | B | 3.0 | 4.8 | 12.3 | 3.1 | 3.8 | 11.4 | 4.1 | 5.0 | b. Linearity/assay reportable range: The linearity of the Accuwell 17-OHP measurement was demonstrated by spiking four serum samples with purified analyte to concentrations of 3.8, 10, 25 and 300 ng/mL. Samples were inter-diluted to make a total of twelve levels with a range between 3.8 - 300 ng/mL. Each sample was combined with lysed, commercially obtained, human, red blood cells to obtain an equivalent hematocrit of 55%. The samples were the spotted onto filter paper and dried. Each sample was tested in quadruplicate for each assay procedure and the average recovery was calculated and plotted against the targeted recoveries. Linear regression of comparison data yielded the following relationship: $$ \text{ON Eluate - } y = 1.0083x - 1.3865 \ r = 0.9998 $$ $$ 3 \text{ Hr Eluate - } y = 1.1015x - 4.7189 \ r = 0.997 $$ $$ \text{ON Direct - } y = 1.1146x - 4.9283 \ r = 0.997 $$ The reportable range for the Accuwell 17-OHP measurements is 3.8-250 ng/mL c. Traceability, Stability, Expected values (controls, calibrators, or methods): The standards and controls are prepared using commercially available human serum stripped of the analyte then purified 17-OHP analyte is added gravimetrically to various concentrations. Each serum concentration is then mixed with lysed red blood cells to a hematocrit concentration equivalent to 55%. The blood mixtures are then spotted onto filter paper in ½ inch circles. The spotted cards are randomly selected throughout the lot and the {5} concentrations tested. Each new lot of cards is tested against a standard curve prepared from the previously approved lots. The stability of the standards and controls were determined by studies that support the expiration date of 12 months. d. Detection limit: The limit of the blank was determined by calculating 2 SD from the mean of n=26 sample aliquots (zero standard) in one run on a single day using each of the three assay procedures. The analytical sensitivity was estimated to be 2.2 ng/mL for the Over night Eluate, 1.5 ng/mL 3 Hour Eluate and 2.3 ng/mL Over Night Direct. e. Analytical specificity: Studies were performed to assess common or known substances that could interfere with the method. Hemoglobin up to 1200 mg/dL, bilirubin, conjugated and unconjugated up to 300 mg/dL, and lipids up to 1875 mg/dL caused no detectable interference. Cross-reactivity was assessed by testing compounds whose structures could potentially cause interference. The lists of compounds tested are listed below: 6 {6} | Substance | Cross Reactivity | Substance | Cross Reactivity | | --- | --- | --- | --- | | 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 3 sulfate | 0.2 % | Cortisone 21-Sulphate Sodium Salt | <0.01 % | | Cortisol | 0.06 % | Dehydroisoandrosterone | <0.01 % | | Desoxycorticosterone | 0.06 % | Dehydroepiandrosterone Sodium Sulphate | <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-Hydroxypregnenolone | <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 % | p e f. Assay cut-off: See expected values below. 2. Comparison studies: a. Method comparison with predicate device: 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 {7} 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 as positive for CAH.) 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 Table 9: Predicate vs. Accuwell O/N Direct | 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 | <43.8 N=130 | 114 | 16 | 130 | Normal | <43.8 N=130 | 112 | 18 | 130 | Normal | <43.8 N=130 | 100 | 30 | 130 | | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=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 Values at the 99th, 97.5th, and 95th, Percentiles for a Table 10: Predicate vs. Accuwell O/N Eluate | 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 | <43.8 N=130 | 106 | 24 | 130 | Normal | <43.8 N=130 | 103 | 27 | 130 | Normal | <43.8 N=130 | 101 | 29 | 130 | | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | | Totals: | | 106 | 27 | 133 | Totals: | | 103 | 30 | 133 | Totals: | | 101 | 32 | 133 | Population of Babies ≥2500 gm and 0-1 Days of Age at Sample Collection The Accuwell 17-OHP EIA (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 = {8} 0.9257 Values at the $99^{\text{th}}$ , $97.5^{\text{th}}$ , and $95^{\text{th}}$ Percentiles for a Population of Babies $\geq 2500$ gm and 0-1 Days of Age at Sample Collection Table 11: Predicate vs. Accuwell 3 Hour Eluate | 99th Percentile | | | | | 97.5th Percentile | | | | | 95th Percentile | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | Predicate | | <23.0 | ≥23.0 | Totals: | Predicate | | <21.7 | ≥21.7 | Totals: | Predicate | | <19.8 | ≥19.8 | | Normal | <43.8 N=130 | 104 | 26 | 130 | Normal | <43.8 N=130 | 103 | 27 | 130 | Normal | <43.8 N=130 | 99 | 31 | | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=3 | 0 | 3 | 3 | Follow-Up | ≥43.8 N=3 | 0 | 3 | | Totals: | | 104 | 29 | 133 | Totals: | | 103 | 30 | 133 | 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 Values at the $95^{\text{th}}$ Percentile for a Population of Babies $\geq 2500$ gm and 2-3 Days of Age at Sample Collection Table 12: Predicate vs. Accuwell O/N Direct | 95th Percentile | | | | | | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Predicate | | < 18.5 | ≥ 18.5 | Totals: | | Normal | < 29.2 N=118 | 113 | 5 | 118 | | Follow-Up | ≥ 29.2 N = 15 | 0 | 15 | 15 | | Totals: | | 113 | 20 | 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 {9} Values at the $95^{\text{th}}$ Percentile for a Population of Babies $\geq 2500$ gm and 2-3 Days of Age at Sample Collection Table 13: Predicate vs. Accuwell O/N Eluate | 95th Percentile | | | | | | --- | --- | --- | --- | --- | | | Accuwell | Normal | 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 Values at the $95^{\text{th}}$ Percentile for a Population of Babies $\geq 2500$ gm and 2-3 Days of Age at Sample Collection Table 14: Predicate vs. Accuwell 3 Hour Eluate | 95th Percentile | | | | | | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Predicate | | < 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 {10} Values at the $97.5^{\text{th}}$ and $95^{\text{th}}$ Percentiles for a Population of Babies $\geq 2500$ gm and $\geq 4$ Days of Age at Sample Collection Table 15: Predicate vs. Accuwell O/N Direct | 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 | < 21.9 N=192 | 181 | 11 | 192 | Normal | < 21.9 N=192 | 170 | 22 | 192 | | Follow-Up | ≥ 21.9 N = 5 | 0 | 5 | 5 | Follow-Up | ≥ 21.9 N = 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 $= 7.3 \, \mathrm{ng/mL}$ , with a range of 2.2 to $22.6 \, \mathrm{ng/mL}$ ; while the predicate kit yielded: mean $= 9.5 \, \mathrm{ng/mL}$ , with a range of 2.7 to $31.0 \, \mathrm{ng/mL}$ . The correlation was found to be: $y$ (Predicate) = 1.090 (Accuwell ON Direct) + 1.565, $R = 0.9538$ Values at the $97.5^{\text{th}}$ and $95^{\text{th}}$ Percentiles for a Population of Babies $\geq 2500$ gm and $\geq 4$ Days of Age at Sample Collection Table 16: Predicate vs. Accuwell O/N Eluate | 97.5th Percentile | | | | | 95th Percentile | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Predicate | | < 15.2 | ≥ 15.2 | Totals: | Predicate | | < 13.6 | ≥ 13.6 | Totals: | | Normal | < 21.9 N=192 | 181 | 11 | 192 | Normal | < 21.9 N=192 | 178 | 14 | 192 | | Follow-Up | ≥ 21.9 N = 5 | 1 | 4 | 5 | Follow-Up | ≥ 21.9 N = 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 $= 7.8 \, \mathrm{ng/mL}$ , with a range of 1.4 to $26.5 \, \mathrm{ng/mL}$ ; while the predicate kit yielded: mean $= 9.5 \, \mathrm{ng/mL}$ , with a range of 2.7 to $31.0 \, \mathrm{ng/mL}$ . The correlation was found to be: $y$ (Predicate) = 1.070 (Accuwell ON Eluate) + 1.172, $R = 0.9142$ {11} Values at the $97.5^{\text{th}}$ and $95^{\text{th}}$ Percentiles for a Population of Babies $\geq 2500$ gm and $\geq 4$ Days of Age at Sample Collection Table 17: Predicate vs. Accuwell 3 Hour Eluate | 97.5th Percentile | | | | | 95th Percentile | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Predicate | | < 15.6 | ≥ 15.6 | Totals: | Predicate | | < 14.1 | ≥ 14.1 | Totals: | | Normal | < 21.9N=192 | 176 | 16 | 192 | Normal | < 21.9N=192 | 174 | 18 | 192 | | Follow-Up | ≥ 21.9N = 5 | 0 | 5 | 5 | Follow-Up | ≥ 21.9N = 5 | 0 | 5 | 5 | | Totals: | | 176 | 21 | 197 | Totals: | | 174 | 23 | 197 | The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean $= 8.3$ ng/mL, with a range of 2.2 to $28.4\mathrm{ng / mL}$ ; while the predicate kit yielded: mean $= 9.5$ ng/mL, with a range of 2.7 to $31.0\mathrm{ng / mL}$ . The correlation was found to be: $y$ (Predicate) = 1.024 (Accuwell 3Hr Eluate) + 1.081, $R = 0.9350$ Values at the $99^{\text{th}}$ and $97.5^{\text{th}}$ Percentiles for a Population of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection Table 18: Predicate vs. Accuwell O/N Direct | 99th (100th) Percentile | | | | | 97.5th Percentile | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Predicate | | < 46.0 | ≥ 46.0 | Totals: | Predicate | | < 43.9 | ≥ 43.9 | Totals: | | Normal | < 45.1N=28 | 28 | 0 | 28 | Normal | < 40.3N=26 | 26 | 0 | 26 | | Follow-Up | ≥ 45.1N = 2 | 1 | 1 | 2 | Follow-Up | ≥ 40.3N = 4 | 3 | 1 | 4 | | 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 $47.8\mathrm{ng / mL}$ ; while the predicate kit yielded: mean $= 25.5$ ng/mL, with a range of 9.0 to $65.0\mathrm{ng / mL}$ . The correlation was found to be: $y$ (Predicate) = 1.22 (Accuwell ON Direct) + 1.971, $R = 0.9556$ {12} Values at the $99^{\text{th}}$ and $97.5^{\text{th}}$ Percentiles for a Population of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection Table 19: Predicate vs. Accuwell O/N Eluate | 99th (100th) Percentile | | | | | 97.5th Percentile | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Predicate | | <31.7 | ≥31.7 | Totals: | Predicate | | <28.4 | ≥28.4 | Totals: | | 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 $40.2\mathrm{ng / mL}$ ; while the predicate kit yielded: mean $= 25.5$ ng/mL, with a range of 9.0 to $65.0\mathrm{ng / mL}$ . The correlation was found to be: $y$ (Predicate) = 1.234 (Accuwell ON Direct) + 1.812, $R = 0.8770$ Values at the $99^{\text{th}}$ and $97.5^{\text{th}}$ Percentiles for a Population of Babies 1400 - 2500 gm and 1-6 Days of Age at Sample Collection Table 20: Predicate vs. Accuwell 3Hr Eluate | 99th (100th) Percentile | | | | | 97.5th Percentile | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Accuwell | Normal | Follow-up | | | Accuwell | Normal | Follow-up | | | Predicate | | <33.3 | ≥33.3 | Totals: | Predicate | | <27.7 | ≥27.7 | Totals: | | Normal | <45.1N=28 | 26 | 2 | 28 | Normal | <40.3N=26 | 23 | 3 | 26 | | Follow-Up | ≥45.1N=2 | 1 | 1 | 2 | Follow-Up | ≥40.3N=4 | 2 | 2 | 4 | | Totals: | | 27 | 3 | 30 | Totals: | | 25 | 5 | 30 | The Accuwell 17-OHP EIA (3Hr Eluate) results for this population were: mean $= 19.3$ ng/mL, with a range of 6.6 to $39.2\mathrm{ng / mL}$ ; while the predicate kit yielded: mean $= 25.5$ ng/mL, with a range of 9.0 to $65.0\mathrm{ng / mL}$ . The correlation was found to be: $y$ (Predicate) = 1.174 (Accuwell 3Hr Eluate) + 2.795, R = 0.8421 b. Matrix comparison: Not applicable. {13} 14 3. Clinical studies: a. Clinical Sensitivity: Not applicable. b. Clinical specificity: Not applicable. c. Other clinical supportive data (when a. and b. are not applicable): See the method comparison study above and the expected values below. 4. Clinical cut-off: See expected values below. 5. Expected values/Reference range: In a separate study of 444 infants whose samples were collected for testing at 1-3 days of age and whose birth weights were ≥ 2500 gm, 17-OHP concentrations were measured using each of the Accuwell 17-OHP methods. Samples were collected on S&amp;S filter paper no. 903 and stored for up to 7 months prior to Accuwell testing. Based on the results obtained, the sponsor provides the following guidelines in the labeling for samples taken at 1-3 days of age from babies with birth weights ≥ 2500 gm. The data show the cut-off values at which re-testing or other follow-up should be considered. Accuwell Expected Values at the 95th, 97.5th and 99th Percentiles for a Population of Babies ≥2500 gm and 1-3 Days of Age at Sample Collection | Accuwell O/N Direct | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | 1-3 | 444 | 20.8 | 25.0 | 27.8 | | Accuwell O/N Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | 1-3 | 444 | 19.1 | 21.7 | 23.9 | {14} 15 | Accuwell 3 hr Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | 1-3 | 444 | 19.3 | 21.5 | 24.3 | In the same study, a small sample (n = 91) of infants with birth weights &lt; 2500 gm were tested using the Accuwell 17-OHP assay methods. The study showed the following: Accuwell Expected Values at the 95th, 97.5th and 99th Percentiles for a Population of Babies &lt;2500 gm and 1-3 Days of Age at Sample Collection | Accuwell O/N Direct | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | N | 95th | 97.5th | 99th | | 1-3 | 91 | 33.9 | 43.9 | 46.4 | | Accuwell O/N Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | 1-3 | 91 | 25.7 | 31.7 | 40.9 | | Accuwell 3 Hr Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | 1-3 | 91 | 26.3 | 32.5 | 40.5 | In a study of 391 infants whose samples were collected for testing at ≥ 4 days of age and whose birth weights were ≥ 2500 gm, 17-OHP concentrations were measured using each of the Accuwell 17-OHP methods. Samples were collected on S&amp;S filter paper no. 903 and stored for up to 7 months prior to Accuwell testing. Based on the results obtained, the following guidelines can be given for samples taken at ≥ 4 days of age from babies with birth weights ≥ 2500 gm. The data show the cut-off values at which re-testing or other follow-up should be considered. Accuwell Expected Values at the 95th, 97.5th and 99th Percentiles for a Population of Babies ≥2500 gm and 1-3 Days of Age at Sample Collection | Accuwell O/N Direct | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | ≥ 4 | 391 | 11.5 | 14.8 | 21.1 | {15} | Accuwell O/N Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | ≥4 | 391 | 13.6 | 15.2 | 19.5 | | Accuwell 3 hr Eluate | | 17-OHP (ng/mL serum) | | | | --- | --- | --- | --- | --- | | | | Percentiles | | | | Age (days) | n | 95th | 97.5th | 99th | | ≥4 | 391 | 14.1 | 15.6 | 19.4 | The values shown here should be used only as a guideline, and were calculated based on a specific population of samples as defined by the manufacturer. The sponsor recommends in the labeling that each laboratory should establish its own cut-off values for 17-OHP in its own population. ## N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. ## O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.