ABX MICROS ES 60 CT; (OPEN TUBE MODEL), (CLOSE TUBE MODEL)

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION MEMORANDUM A. 510(k) Number: k141161 B. Purpose for Submission: Clearance of a new device. C. Manufacturer and Instrument Name: Horiba ABX SAS, ABX Micros ES 60 OT and ABX Micros ES 60 CT D. Type of Test or Tests Performed: Quantitative test for white blood count (WBC), red blood count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLT), lymphocyte (LYM)%/#, monocyte (MON)%/#, granulocytes (GRA)%/#, and red cell distribution width (RDW). E. System Descriptions: 1. Device Description: The ABX MICROS ES 60 is a quantitative, automated hematology analyzer and leukocyte differential counter for in vitro diagnostic use in clinical laboratories. The instrument system is comprised of the analyzer and a suite of analytical reagents that allow for simultaneous quantitative determination of hemoglobin measurement, cell counting, quality control, calibration, and cleaning. The system is a microprocessor controlled hematology analyzer used for the in vitro diagnostic testing of whole blood specimens. It operates in complete blood count (CBC) and Differential (DIFF) mode using a combination of focused flow impedance and light transmission technologies. It is available in Closed (CT) or Open Tube (OT) sampling versions. 2. Principles of Operation: The ABX Micros ES 60 principle of automated cell counting and sizing is used in the analysis of the whole blood. Each cell suspended in a conductive liquid (diluent) acts as an insulator. As each cell goes through the aperture, it momentarily increases the resistance of the electrical path between two submerged electrodes on either side of the aperture. This causes a measurable electronic pulse. While the number of pulses indicates particle count, the amplitude of the electrical pulse is proportional to the cell volume. These pulses are sent to the Signal Conditioner for analog to digital conversion. Pulse counts and digitized pulse measurements are sent to the System Manager for processing by the algorithms where the reported parameter values, flags and histograms are generated. {1} The lysing reagent breaks down the RBC cell membrane and releases the hemoglobin contained by the cell. The hemoglobin, released by the lysing reagent, combines with the potassium cyanide from the lysing reagent to form a chromogenous cyanmethemoglobin compound. This compound is then measured through the optical part of the WBC/HGB chamber by spectrophotometry at a wavelength of 550 nm. 3. Modes of Operation: Does the applicant’s device contain the ability to transmit data to a computer, webserver, or mobile device? Yes ☐ X ☐ or No ☐ Does the applicant’s device transmit data to a computer, webserver, or mobile device using wireless transmission? Yes ☐ or No ☐ X ☐ 4. Specimen Identification: Specimen identification is by manual sample identification or with the use of a hand held barcode scanner. 5. Specimen Sampling and Handling: Two models of ABX Micros ES 60 are available. - The Closed Tube (CT) instrument: Cap-piercing mechanism; the blood collection tube is placed directly in the analyzer without removing the cap. - The Open Tube (OT) instrument: The cap from the blood collection tube must be removed before analyzing the sample. 6. Calibration: ABX Minocal calibrator (k955925) is used for the ABX Micros ES 60 calibration procedure. Assigned assay values are traceable to reference methods. Calibration is performed by a HORIBA ABX SAS representative during specific situations such as installation, maintenance or service intervention. 7. Quality Control: ABX Minotrol 16 Control (high, normal and low) (k850755) is used to monitor system performance for all directly measured and calculated CBC and Diff parameters. Assigned assay values are determined on validated instruments using the appropriate reagents. 8. Software: {2} FDA has reviewed applicant’s Hazard Analysis and Software Development processes for this line of product types: Yes ☐ X ☐ or No ☐ ## F. Regulatory Information: 1. Regulation section: CFR 864.5220, Automated differential cell counter 2. Classification: Class II 3. Product code: GKZ, Counter, Differential Cell 4. Panel: Hematology (81) ## G. Intended Use: 1. Indication(s) for Use: The ABX MICROS ES 60 (OT and CT models) is a quantitative multi-parameter, automated hematology analyzer for in vitro diagnostic use in clinical laboratories to identify and enumerate the following parameters: WBC, RBC, HGB, HCT, MCV, MCH, MCHC, RDW, PLT, MPV, LYM%, LYM#, MON%, MON#, GRA%, GRA#, in K₂EDTA and K₃EDTA anticoagulated venous whole blood samples of adult patient population. It is not intended to be used for pediatric subjects. 2. Special Conditions for Use Statement(s): For prescription use only ## H. Substantial Equivalence Information: 1. Predicate Device Name(s) and 510(k) numbers: Horiba ABX Micros 60, k030799 {3} # 2. Comparison with Predicate Device: | Similarities | | | | --- | --- | --- | | Item | Device (ABX MICROS ES 60) | Predicate (ABX MICROS 60 - k030799) | | Intended Use | The ABX MICROS ES 60 (OT and CT models) is a quantitative multi-parameter, automated hematology analyzer for in vitro diagnostic use in clinical laboratories to identify and enumerate the following parameters: WBC, RBC, HGB, HCT, MCV, MCH, MCHC, RDW, PLT, MPV, LYM%, LYM#, MON%, MON#, GRA%, GRA#, in K2EDTA and K3EDTA anticoagulated venous whole blood samples of adult patient population. It is not intended to be used for pediatric subjects. | The ABX MICROS 60 Hematology Analyzer is a fully automated (microprocessor controlled) hematology analyzer used for the in vitro diagnostic testing of whole blood specimens or blood cell concentrates. It operates in complete blood count (CBC) mode. | | Principles of Measurement | | | | RBC, PLT, HCT, MPV | Impedance | Same | | HGB | Spectrophotometry | Same | | WBC, WBC Differential (LYM, MON, GRA)) | Impedance | Same | | RDW, MCV, MCH, MCHC | Calculation | Same | | Reagents | | | | Diluent | ABX Minidil LMG | Same | | Lyse | ABX Minilyse LMG or ABX Alphalyse 360 | Same | | Cleaner | ABX Miniclean | Same | | Reagent Pack | ABX Minipack LMG | Same | | Concentrated cleaning reagent | ABX Minoclair | Same | | Quality Controls | ABX Minotrol 16 (3 levels) | Same | | Calibrator | ABX Minocal | Same | | System configuration | Bench top Handheld barcode reader (optional) Integrated barcode reader (CT version only) Printer | Same | | Sampling mechanism | Single tube presentation – open and closed vial sampling | Same | | Aspiration pathway | Single sampling probe and common aspiration pathway used for all sample presentation modes | Same | | Minimum sample volume Specimen sample volume | 50 μL 10 μL | Same | | Counting aperture diameters RBC/PLT WBC | 50 μm 80 μm | Same | {4} | Similarities | | | | --- | --- | --- | | Item | Device (ABX MICROS ES 60) | Predicate (ABX MICROS 60 - k030799) | | Dilution ratios: RBC/PLT chamber | 1/15000 | Same | | Differences | | | | --- | --- | --- | | Item | Device (ABX MICROS ES 60) | Predicate (ABX MICROS 60 - k030799) | | User Interface Display | Automated instrument with 8” LCD touch screen display | Automated instrument with 3” LCD display | | Software application | Linux-based software application | Internally developed software application | | Analytical cycle | 1. Draining sequence done by vacuum 2. No air bubble. | 1. Draining sequence movement 2. Presence of air bubble at the end of all cycles | | Sample types | Whole blood samples only | Whole blood samples and blood cell concentrates | | Dilution ratio: WBC chamber | 1/260 | 1/250 | | Throughput | OT/CT models: 60/50 samples per hour | OT/CT model: approx. 60/55 samples per hour | # I. Special Control/Guidance Document Referenced (if applicable): CLSI EP05-A2: Evaluation of Precision Performance of Quantitative Measurement Methods - 2004 CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedure: A Statistical Approach - 2003 CLSI EP07-A2: Interference Testing in Clinical Chemistry - 2005 CLSI EP09-A3: Measurement Procedure Comparison and Bias Estimation Using Patient Samples - 2013 CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures - 2012 CLSI EP28-A3c: Defining, Establishing and Verifying Reference Intervals in the Clinical Laboratory - 2008 CLSI H20-A2: Reference Leukocyte (WBC) Differential Count (Proportional) and Evaluation of Instrumental Methods - 2007 CLSI H26-A2: Validation, Verification and Quality Assurance of Automated Hematology Analyzers - 2010 # J. Performance Characteristics: {5} # 1. Analytical Performance: # a. Method Comparison # 1. Comparability between ABX Micros ES 60 CT Vs. ABX Micros ES 60 OT A total of 237 whole blood specimens from adult patients were analyzed at one test site in France. Each sample was analyzed in duplicate on the ABX Micros ES 60 CT (Close Tube model) and on the ABX Micros ES 60 OT (Open Tube model). Bias was estimated at three points for each parameter: the low end of the distribution of observations, the mid-point, and the high end of the distribution. Bias was estimated separately for each replicate. Acceptance criteria were met for all measurands at all levels. These findings support the claim that both Open Tube and Close Tube models of ABX Micros ES60 device yield comparable performances across the analytical range for all parameters. Passing Bablok regression analyses demonstrate comparable performance between Micros ES 60 OT and Micros ES 60 CT models across the analytical range for all the parameters. Bias data for the first replicate of samples tested in the comparison study are provided below. First replicate: | | Passing Bablock Regression | | | Bias Claim | %Bias confidence interval 95% | | | | --- | --- | --- | --- | --- | --- | --- | --- | | | r2 | Slope | Intercept | | Level 1 | Level 2 | Level 3 | | WBC | 0.998 | 1.00 | 0.08 | 9.80% | [-3.4%; 6.5%] | [0.1%; 1.7%] | [0%; 1.6%] | | RBC | 0.996 | 1.00 | -0.02 | 3.40% | [-2.8%; 2.7%] | [-1.1%; 0.8%] | [-1%; 0.6%] | | HGB | 0.997 | 0.99 | 0.32 | 3.10% | [-1.4%; 7%] | [-0.7%; 2.3%] | [-1.1%; 1.5%] | | HCT | 0.996 | 0.98 | 0.59 | 1.70% | [-1.5%; 1.7%] | [-1.4%; 0.6%] | [-1.8%; 0.2%] | | MCV | 0.975 | 1.00 | 0 | 1.20% | [-5.2%; 0%] | [-3.3%; 0%] | [-2.7%; 0.6%] | | MCH | 0.948 | 1.07 | -1.69 | | [-4.6%; 5.6%] | [-1.9%; 5.4%] | [-1%; 6.4%] | | MCH | 0.706 | 1.33 | -10.57 | | [-24.8%; 9.8%] | [-11.8%; 13%] | [-8.3%; 17.2%] | | RDW | 0.896 | 1.00 | 0.20 | 4.00% | [-8.6%; 7.9%] | [-6.1%; 6.1%] | [-4.6%; 5.3%] | | PLT | 0.987 | 0.98 | 5.63 | 14.7% | [-12.8%; 28.3%] | [-3.2%; 3.7%] | [-3.6%; 0.8%] | | MPV | 0.955 | 1.00 | -0.20 | 2.30% | [-3.3%; 3.0%] | [-2.4%; 1.00%] | [-2.7%; 0.7%] | | LYM | 0.987 | 1.00 | 0.30 | 12.0% | [-15.5%; 43.7%] | [-0.2%; 2.1%] | [-0.6%; 1.7%] | | LYM | 0.999 | 1.00 | 0.10 | 12.0% | [0.6%; 10%] | [0.9%; 4.6%] | [0.7%; 3.9%] | | MON | 0.960 | 1.03 | 0.10 | 26.0% | [-6.7%; 16%] | [0.4%; 6.8%] | [0.2%; 6.6%] | | MON | 0.979 | 1.00 | 0.00 | 13.2% | [0%; 0%] | [0%; 0%] | [0%; 0%] | | GRA | 0.989 | 1.00 | -0.73 | 9.10% | [-10.9%; 3.0%] | [-3.4%; 0.8%] | [-2.0%; 0.7%] | | GRA | 0.999 | 1.00 | 0.00 | 9.10% | [0%; 0%] | [0%; 0%] | [0%; 0%] | # 2. ABX Micros ES (CT) and ABX Micros ES (OT) vs. Manual Microscopy 100 normal and 100 pathological samples preserved in $\mathrm{K}_2\mathrm{EDTA}$ covering the full analytic range of the ABX Micros ES60 were analyzed in duplicate on the ABX Micros ES 60 OT and ABX Micros ES 60 CT. Two slides for each sample were prepared using May Grunwald Giemsa staining. Samples were classified into Normal or Abnormal samples for the ABX Micros ES 60 OT, ABX Micros ES 60 {6} CT, and the reference method (manual slide microscopy for differential count). Method comparison studies were designed using the CLSI H20-A2 procedure by the construction of the "envelope" (or Confidence Interval CI) that takes into account the imprecision of both the test and the manual method (95% and 99% confidence interval, respectively). Based on a Gaussian distribution, 5% of the individual results may fall outside the 95% envelope. No result should exceed the 99% limits without biological explanation and a flag. Samples exceeding the 99% limits with a biological explanation and a flag are considered as outliers. Evaluation of clinical sensitivity/specificity was based on comparison results between the slide microscopy and the ABX Micros ES 60 analyzer using 200 samples as follows: | a. Parameter | ABX Micros ES 60 OT | | ABX Micros ES 60 CT | | | --- | --- | --- | --- | --- | | | Number / Percentage of samples outside the 95% CI | Number / Percentage of samples outside the 99% CI | Number / Percentage of samples outside the 95% CI | Number / Percentage of samples outside the 99% CI | | LYM% | 3#/ 1.5% | 0# 0% | 7# / 3.5% | 0# 0% | | MON% | 5#/ 2.5% | 0# 0% | 5# / 2.5% | 0# 0% | | GRA% | 2#/ 1% | 0# 0% | 9# / 4.5% | 0# 0% | Samples with a quantitative or a morphological abnormality observed on the reference method (slide count) or on the predicate Micros 60 were compared with the quantitative flag on the Micros ES60 OT and CT. | | Micros ES 60 OT | | Micros ES 60 CT | | Micros 60 | | | | --- | --- | --- | --- | --- | --- | --- | --- | | | | Positive (Abnormal) | Negative (Normal) | Positive (Abnormal) | Negative (Normal) | Positive (Abnormal) | Negative (Normal) | | Reference Method | Positive (Abnormal) | 97 | 3 | 96 | 4 | 99 | 1 | | | Negative (Normal) | 43 | 57 | 51 | 49 | 66 | 34 | | | Micros ES 60 OT | Micros ES 60 CT | Micros 60 | | --- | --- | --- | --- | | % Efficiency | 77.0% | 72.5% | 66.5 | | % Sensitivity | 97.0% | 96.0% | 99.0 | | 95% CI Sensitivity | 91.5% to 99.0% | 90.2% to 98.4% | 94.6 to 99.8% | | % Specificity | 57.0% | 49.0% | 34.0 | | 95% CI Specificity | 47.2% to 66.3% | 39.4 to 58.7 % | 25.5 to 43.7% | | % | 69.3% | 65.3% | 60.0 | | % | 95.0% | 92.5% | 97.1 | PPV = Positive Predictive Value NPV = Negative Predictive Value {7} 3. Comparability between ABX Micros ES 60 and ABX Micros 60 A total of 179 whole blood specimens from adult patients were analyzed at four test sites in the US. Each of the samples was analyzed in duplicate on the ABX Micros ES 60 (CT model) and on the predicate ABX Micros 60. Bias was estimated at three points for each parameter: the low end of the distribution of observations, the mid-point, and the high end of the distribution. Bias was estimated separately for each replicate. Acceptance criteria were met for all measurands at all levels. These findings support the claim that the ABX Micros ES60 candidate device and the ABX Micros 60 predicate device are substantially equivalent, and demonstrate acceptable levels of bias. Method Comparison (ABX Micros 60 vs. ABX Micros ES 60 CT), Estimated bias for combined sites. | Parameter | N | R² | Slope | Lower Limit | Higher Limit | Intercept | Lower Limit | Higher Limit | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | WBC (10³/mm³) | 174 | 0.999 | 1.00 | 0.99 | 1.00 | 0.00 | 0.00 | 0.07 | | RBC (10⁶/mm³) | 174 | 0.989 | 0.97 | 0.96 | 0.99 | 0.065 | 0.009 | 0.120 | | HGB (g/dL) | 179 | 0.991 | 0.98 | 0.97 | 1.00 | 0.15 | 0.00 | 0.33 | | HCT (%) | 172 | 0.988 | 0.98 | 0.96 | 0.99 | 0.48 | -0.02 | 1.06 | | MCV (μm³) | 179 | 0.982 | 1.00 | 1.00 | 1.00 | 0.00 | 0.00 | 0.00 | | MCH | 179 | 0.924 | 1.03 | 1.00 | 1.08 | -0.77 | -2.23 | 0.30 | | MCHC | 179 | 0.59 | 1.11 | 1.00 | 1.25 | -3.36 | -7.93 | 0.30 | | RDW (%) | 179 | 0.944 | 0.91 | 0.87 | 0.94 | 1.65 | 1.15 | 2.15 | | PLT (10³/mm³) | 171 | 0.995 | 0.98 | 0.97 | 0.99 | -2.97 | -7.90 | 1.79 | | MPV (μm³) | 179 | 0.808 | 1.05 | 1.00 | 1.10 | -0.39 | -0.82 | 0.00 | | LYM (%) | 179 | 0.917 | 0.97 | 0.95 | 0.99 | 0.24 | -0.18 | 0.71 | | LYM # (10³/mm³) | 179 | 0.996 | 0.99 | 0.95 | 1.00 | 0.01 | 0.00 | 0.06 | | MON (%) | 179 | 0.778 | 1.11 | 1.07 | 1.15 | -0.09 | -0.35 | 0.26 | | MON # (10³/mm³) | 179 | 0.961 | 1.00 | 1.00 | 1.03 | 0.10 | 0.07 | 0.10 | | GRA (%) | 179 | 0.913 | 1.01 | 0.99 | 1.03 | -1.35 | -2.65 | -0.14 | | GRA # (10³/mm³) | 179 | 0.998 | 1.00 | 0.99 | 1.00 | 0.00 | 0.00 | 0.02 | Correlation coefficient; R² Criteria &gt;0.95 for WBC, RBC, HGB, HCT, PLT {8} To establish that the candidate device (ABX Micros ES 60 CT) is comparable with the predicate device (ABX Micros 60) following bias and $r^2$ acceptance criteria was established for Rep 1 and Rep 2. Bias Acceptance Criteria | Parameter | +/- % Bias Criteria | R2Criteria | | --- | --- | --- | | WBC | 9.8 | >0.95 | | RBC | 3.4 | >0.95 | | HGB | 3.1 | >0.95 | | HCT | 1.7 | >0.95 | | MCV | 1.2 | - | | RDW | 4.0 | - | | PLT | 14.7 | >0.95 | | MPV | 2.3 | - | | LYM% | 12 | - | | LYM# | 12 | - | | MON% | 26 | - | | MON# | 13.2 | - | | GRA% * | 9.1 | - | | GRA# * | 9.1 | - | * Specific to Neutrophils # b. Precision/Reproducibility: # Repeatability Study Repeatability was performed in-house using 12 normal and 10 abnormal fresh whole blood samples collected into tubes containing $\mathsf{K}_2\mathsf{EDTA}$ anticoagulant. Each sample was run 10 consecutive times on two models of the Micros ES 60 (ES 60 OT and ES 60 CT), in a single day and all runs were completed within 8 hours of sample collection. The standard deviation was compared with the mean value of each parameter and the CV% was calculated. Results were within the specifications shown below. Precision (Repeatability) Acceptance Criteria | | WBC | RBC | HGB | HCT | PLT | LYM% | MON% | GRA% | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | %CV | <2.5 | <2 | <1.5 | <2 | <5 | <10 | <20 | <4 | # Reproducibility Reproducibility was assessed on three Micros ES 60 instruments at three sites, one operator per site. At each site; High, Normal, and Low levels of one single lot of {9} Minotrol control material were run in duplicate, twice each day, for a total of 82 days among the three sites. Total standard deviation and CV% were calculated for each measurand and results obtained were within specifications shown below. The Micros ES 60 met the acceptance criteria. Reproducibility Study – Combined Sites | All Sites Combined | | | | Within-Run | Between-Day | Between-Run | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Parameter | Control Level | N | Mean | CV% | CV% | CV% | Total CV% | CV% Claim | | WBC (x 10³/mm³) | Low | 332 | 2.05 | 2.78 | 1.84 | 1.76 | 3.77 | 7.00 | | | Normal | 340 | 7.47 | 1.78 | 1.24 | 0.00 | 2.17 | 5.00 | | | High | 328 | 20.14 | 1.11 | 1.38 | 0.56 | 1.86 | 4.00 | | RBC (x 10⁶/mm³) | Low | 332 | 2.38 | 1.30 | 1.20 | 0.41 | 1.82 | 4.00 | | | Normal | 340 | 4.67 | 1.11 | 1.16 | 0.36 | 1.65 | 3.00 | | | High | 328 | 5.71 | 1.01 | 1.33 | 0.55 | 1.76 | 3.00 | | HGB (g/dL) | Low | 332 | 6.01 | 1.13 | 0.96 | 0.62 | 1.60 | 5.00 | | | Normal | 340 | 13.52 | 0.91 | 0.94 | 0.37 | 1.36 | 4.00 | | | High | 328 | 17.90 | 0.74 | 1.09 | 0.49 | 1.40 | 3.00 | | HCT (%) | Low | 332 | 16.62 | 1.35 | 2.21 | 0.55 | 2.65 | 5.00 | | | Normal | 340 | 38.11 | 1.15 | 1.62 | 0.59 | 2.07 | 4.50 | | | High | 328 | 51.21 | 1.07 | 1.93 | 0.58 | 2.28 | 4.00 | | MCV (μm³) | Low | 332 | 69.90 | 0.49 | 2.02 | 0.42 | 2.12 | 4.00 | | | Normal | 340 | 81.70 | 0.35 | 1.51 | 0.35 | 1.59 | 3.00 | | | High | 328 | 89.68 | 0.38 | 1.72 | 0.34 | 1.79 | 2.50 | | PLT (10³/mm³) | Low | 332 | 75.64 | 5.99 | 5.15 | 3.29 | 8.56 | 15.00 | | | Normal | 340 | 264.63 | 2.96 | 1.29 | 0.46 | 3.26 | 10.00 | | | High | 328 | 521.83 | 2.13 | 1.72 | 0.53 | 2.78 | 7.00 | | RDW (%) | Low | 332 | 13.42 | 1.86 | 2.94 | 0.74 | 3.56 | 5.0 | | | Normal | 340 | 12.96 | 1.31 | 3.18 | 0.84 | 3.55 | 5.0 | | | High | 328 | 12.30 | 1.26 | 2.92 | 0.91 | 3.31 | 5.0 | | MCH (10³/mm³) | Low | 332 | 25.28 | 1.00 | 1.25 | 0.44 | 1.66 | 7.0 | | | Normal | 340 | 28.97 | 0.74 | 1.25 | 0.32 | 1.49 | 4.5 | | | High | 328 | 31.35 | 0.72 | 1.22 | 0.27 | 1.44 | 4.0 | | MCHC (%) | Low | 332 | 36.16 | 1.04 | 2.14 | 0.23 | 2.39 | 7.0 | | | Normal | 340 | 35.48 | 0.78 | 1.91 | 0.36 | 2.09 | 4.5 | | | High | 328 | 34.97 | 0.80 | 2.12 | 0.00 | 2.26 | 4.0 | | LYM (%) | Low | 332 | 60.80 | 1.73 | 0.00 | 1.05 | 2.02 | 8.00 | | | Normal | 340 | 32.34 | 1.66 | 1.80 | 0.19 | 2.45 | 8.00 | | | High | 328 | 15.50 | 1.70 | 5.25 | 1.43 | 5.70 | 8.00 | {10} | All Sites Combined | | | | Within-Run | Between-Day | Between-Run | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Parameter | Control Level | N | Mean | CV% | CV% | CV% | Total CV% | CV% Claim | | MON (%) | Low | 332 | 11.18 | 6.30 | 3.95 | 0.00 | 7.43 | 15.00 | | | Normal | 340 | 7.64 | 4.11 | 2.77 | 1.27 | 5.11 | 15.00 | | | High | 328 | 6.56 | 4.06 | 9.28 | 6.55 | 12.06¹ | 11.00 | | GRA % | Low | 332 | 27.99 | 3.45 | 1.65 | 2.48 | 4.56 | 12.00 | | | Normal | 340 | 60.02 | 0.98 | 1.05 | 0.00 | 1.44 | 4.00 | | | High | 328 | 77.94 | 0.47 | 0.90 | 0.45 | 1.11 | 3.00 | | LYM# | Low | 332 | 1.20 | 3.61 | 2.21 | 2.00 | 4.68 | 8.00 | | | Normal | 340 | 2.37 | 2.57 | 3.02 | 0.23 | 3.97 | 8.00 | | | High | 328 | 3.07 | 2.27 | 6.68 | 0.84 | 7.29 | 8.00 | | MON# | Low | 332 | 0.20 | 10.09 | 5.97 | 0.00 | 11.73 | 15 | | | Normal | 340 | 0.52 | 6.85 | 3.60 | 2.56 | 8.15 | 14 | | | High | 328 | 1.27 | 4.85 | 9.45 | 7.04 | 12.74¹ | 11 | | GRA# | Low | 332 | 0.66 | 8.23 | 0.74 | 3.13 | 8.83 | 12.0 | | | Normal | 340 | 4.59 | 2.15 | 0.28 | 0.29 | 2.19 | 4.0 | | | High | 328 | 15.79 | 1.24 | 0.86 | 0.63 | 1.63 | 3.0 | ¹: Exceeds acceptance criteria limits; however, these results support the claim of reproducibility for low, normal and high levels of controls with the overall reproducibility claim being met in 97% of the tests. ## c. Linearity: Linearity was assessed using R&amp;D Systems CBC-LINE high and low range linearity kits according to the manufacturer's instructions. The expected values of the kit samples were considered the "true values". Each level was run in replicates of four (n=4) as recommended by the kit supplier. For each level, the 4 replicate results were plotted versus the theoretical value. The findings of the polynomial regression analysis indicate that the ABX Micros ES 60 exhibits linearity across the claimed range. The Micros ES 60 met the acceptance criteria. Analytical Measuring Range | Parameter | AMR on Micros ES60 | | --- | --- | | WBC (10³/mm³) | 0.8 – 100 | | RBC (10⁶/mm³) | 0.7 – 8 | | PLT (10³/mm³) | 42 – 2200 | | HGB (g/dL) | 0.6 - 24.0 | | HCT (%) | 8.0 - 70.0 | ## d. Carryover: The potential for sample carryover was tested in duplicate on the ABX Micros ES 60 OT and CT instruments using alternating high and low sample concentrations. Carry-over effects were evaluated by assaying a diluted sample 3 times, low (L1-3) {11} then a sample with high cell concentrations 3 times (H1-3), followed immediately by testing a diluted sample consecutively 3 times (L4-6). Clinical samples were either concentrated or diluted to obtain the following target high and low sample values: | | High Samples | Low Samples | | --- | --- | --- | | WBC | 40 to 80 x10^3/mm^3 | 0.8 to 1.2 x 10^3/mm^3 | | RBC | 6 to 9 x 10^6/mm^3 | 1 to 2 x10^6/mm^3 | | HGB | 15 to 20 g/dL | 5 to 7 g/dL | | PLT | 500 to 1000 x10^3/mm^3 | 15 to 50 x10^3/mm^3 | All carry-over results were within specifications, &lt;1% for the ABX Micros ES OT and CT Systems for the following parameters: WBC, RBC, HGB, and PLT. e. Interfering Substances: The study to evaluate interference effect followed two methodologies: By addition: The evaluation of the effect of potentially interfering substances added to the sample of interest. A potential interfering substance was added to a sample and the bias relative to a control portion of the sample was evaluated ("paired-difference testing"). This bias was compared to the acceptance criteria. For all tests performed with potential interferents, the bias remained below the acceptable limit. Therefore, no significant interference was observed for urea, bilirubin, lipemia, and hemolysis. Interference of WBC counts has been observed in presence of yeast in the sample. By comparison: Evaluation of the bias of individual specimen. Representative patient specimens and a control sample (without interferent) were run in duplicate using the Micors ES 60 versus the reference device, Micros 60, to obtain a comparative measurement. The bias versus comparative measurement values was plotted for each specimen group. Both measurement procedures had 10 to 20 samples in each group to demonstrate sufficient precision. A comparable effect was observed on the ABX Micros ES 60 and the reference device when interference was tested for the following: WBC fragments, myelocytes, nucleated RBC, RBC inclusion, RBC agglutinins / cold agglutinins / RBC rouleaux, dual RBC population, RBC fragments, target cells, platelet aggregates, platelet satellitism, macrothrombocytes / large platelets, small RBC leukocytosis. Interference of megakaryocytes with WBC and parasites with monocyte counts could not be tested, but that are well described in literature. 2. Other Supportive Instrument Performance Data Not Covered Above: 1. Specimen Stability Studies: Eleven (11) whole venous blood specimens (collected in K₂EDTA and K₃EDTA) were analyzed on the ABX Micros ES 60 (OT model) at one site in France in order to cover the AMR for the majority of measurands. Following the collection (T0), each specimen was divided in half, with one sample stored at ambient temperature (20- {12} $24^{\circ}\mathrm{C})$ and the other stored under refrigerated conditions $(2 - 8^{\circ}\mathrm{C})$ . Testing for stability was performed at 2, 6, 8, 10, 24, 36, 48, 60 and 72 hours after T0. The first aliquot of each specimen was run sequentially in any order. The second aliquot was run in reverse order to minimize the effects of carryover and drift. The acceptance criteria for sample stability is given as an acceptable maximum bias of the value at T with the value at T0. All data met specifications. | Parameters | Sample stability when stored refrigerated (2-8°C) | Sample stability when stored at room temperature (20-24°C) | | --- | --- | --- | | WBC, RBC, HGB, HCT, MCV, MCH, MCHC, PLT | 48 hours | 36 hours | | RDW | 36 hours | 10 hours | | MPV | 24 hours | 24 hours | | LYM%, LYM#, MON%, MON#, GRA%, GRA# | 24 hours | 8 hours | # 2. Comparability between Anticoagulants A total of 52 normal and pathological blood specimens were analyzed on the ABX Micros ES 60 (CT model) at three sites in the US. Testing was performed using a different instrument and operator at each site. The specimens used in this study were venous blood specimens that were prospectively specifically collected for this study. Each subject provided blood collected in both $\mathrm{K}_2\mathrm{EDTA}$ and $\mathrm{K}_3\mathrm{EDTA}$ . Each of the samples was analyzed in duplicate on the ABX Micros ES 60 CT. Bias was estimated at three points for each parameter: the low end of the distribution of observations, the mid-point, and the high end of the distribution. Bias was estimated separately for each replicate. Acceptance criteria were met for all measurands at all levels. These findings support the claim that $\mathrm{K}_2\mathrm{EDTA}$ and $\mathrm{K}_3\mathrm{EDTA}$ specimens give comparable results as measured on the ABX Micros ES 60 hematology analyzer. # 3. Determination of limit of blank, lower limits of detection and quantitation: # Verification of Limit of Blank (LoB) Plasma samples, obtained by centrifugation of normal samples, were used as blank samples, in order to be as close as possible as the blood sample matrix. To estimate the LoB, a total of 60 repeated measurements of different plasma were run in the same series (5 different samples run 12 times). This test is performed on 2 instruments using two reagents lots. Results on Micros ES 60 OT and Micros ES60 CT were similar and met specifications. LoB obtained from 60 repeated measurements of 5 different plasma samples: | Measurand | LoB | | --- | --- | | WBC | 0.1x103/mm3 | | RBC | 0.01x106/mm3 | | HCT | 0.1 % | | PLT | 1x103/mm3 | {13} # Verification of Limit of Detection (LoD) A set of six samples with very low parameter concentration (i.e. in the range of LoB and $4 \times$ LoB) were run 10 times over several days. LoD was estimated from the calculation of the pooled standard deviation of 60 results. Testing was performed on two (2) instruments with two reagents lots. Results on Micros ES 60 OT and Micros ES 60 CT were similar and met specifications. Testing from 10 runs of 6 low samples on each instrument resulted in the following results for LoD: | Measurand | LoD | | --- | --- | | WBC | 0.2x103/mm3 | | RBC | 0.01x106/mm3 | | HGB | 0.5 g/dL | | HCT | 0.2 % | | PLT | 4x103/mm3 | # Verification of Limit of Quantitaion (LoQ) To estimate the limit of quantitation, several ranges of linearity in low concentrations were prepared. Between 3 to 6 samples by concentratin level were prepared and run at least 5 times each using 40 replicates per level. The LoQ data are considered acceptable when the %Total-error is smaller than the desired total error for each measurand. This test is performed on two (2) instruments with two reagents lots. Results on Micros ES 60 OT and Micros ES 60 CT were similar and passed acceptance limits. Testing from 40 runs of 4 samples on each instrument resulted in the following results for LoQ: | Measurand | LoQ | | --- | --- | | WBC | 0.8x103/mm3 | | RBC | 0.7x106/mm3 | | HGB | 0.6 g/dL | | HCT | 8% | | PLT | 42x103/mm3 | # 4. Reference Intervals: # Verification of Adult Reference Intervals Adult reference intervals were assessed on a total of 275 (135 male and 140 female) normal adult samples (whole blood samples collected in $\mathrm{K}_2\mathrm{EDTA}$ ). Samples were analyzed in duplicate on the ABX Micros ES 60 (OT model) and ABX Micros ES 60 (CT model) at one test site in the US. The nonparametric data analysis method was used, depending only on the ranks of the reference data arranged in order of increasing size. Per EP28-A3, the reference interval is determined to be between and including the lower and upper reference limits, which enclose $95\%$ of the values from the reference population subjects. Confidence intervals for the reference limit were calculated using a $90\%$ probability ( $90\%$ CI). {14} Defined reference values are not significantly different between the ABX Micros ES 60 OT and CT models. For each gender, a single reference interval applicable (Shared Interval) on both versions of the analyzers has been defined as described in the following table. | MICROS ES60 Reference Interval | MALES (N=135) | | FEMALES (N=140) | | | --- | --- | --- | --- | --- | | | LOW | HIGH | LOW | HIGH | | WBC (103/mm3) | 4.3 | 9.6 | 4.2 | 10.3 | | RBC (106/mm3) | 4.1 | 5.7 | 4.0 | 5.1 | | HGB (g/dl) | 12.6 | 16.7 | 11.6 | 15.1 | | HCT (%) | 38.3 | 50.8 | 35.8 | 46.4 | | MCV (μm3) | 83 | 97 | 83 | 98 | | MCH (pg) | 26.7 | 32.3 | 26.8 | 32.5 | | MCHC (g/dl) | 31.7 | 34 | 31.8 | 34.0 | | RDW (%) | 11.1 | 14.4 | 11.3 | 13.9 | | PLT (103/mm3) | 156 | 370 | 181 | 393 | | MPV (μm3) | 6.3 | 9.1 | 6.5 | 9.0 | | LYM (103/mm3) | 1.1 | 3.1 | 1.2 | 3.4 | | MON (103/mm3) | 0.1 | 0.6 | 0.1 | 0.6 | | GRA (103/mm3) | 2.6 | 7.0 | 2.7 | 7.4 | | LYM (%) | 16.5 | 44.7 | 17.7 | 45.2 | | MON (%) | 3.1 | 8.3 | 3.3 | 8.1 | | GRA (%) | 49.1 | 76.9 | 49.2 | 77.7 | # K. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. # L. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.