(1) Immunoassay for the in vitro qualitative detection of IgM antibodies to CMV in human serum, lithium-heparin plasma, K2-EDTA plasma, and K3-EDTA plasma. The test is intended as an aid in the diagnosis of recent or current CMV infection in individuals for which a CMV IgM test was ordered, including pregnant women. Performance characteristics have not been evaluated in immunocompromised or immunosuppressed individuals. This test is not intended for use in neonatal screening or for use at point of care facilities. This assay is not intended for use in screening blood and plasma donors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys immunoassay analyzers.

(2) PreciControl CMV IgM is used for quality control of the Elecsys CMV IgM immunoassay on the Elecsys and cobas e immunoassay analyzers.

Device Description

(1) Elecsys CMV IgM is a u-capture immunoassay with streptavidin microparticles, biotinylated recombinant CMV-specific antigen labeled with a ruthenium complex and electrochemiluminescence detection. The results are determined using a calibration curve which is instrument-specifically generated by a 2-point calibration and a master curve provided via the reagent bar code. The test system contains the human serum-based calibrators intended for use with the system.

The CMV IgM assay begins with an automatic 1:20 predilution of sample with Elecsys Diluent Universal and the addition of biotinylated monoclonal anti-h-IgM-specific antibodies.

During the second incubation, CMV-specific recombinant antigen labeled with a ruthenium complex and streptavidin-coated microparticles are added. Anti-CMV IgM antibodies present in the sample react with the ruthenium-labeled CMV-specific recombinant antigen. The complex becomes bound to the solid phase via interaction of biotin and streptavidin.

The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell. Application of a voltage to the electrode induces chemiluminescent emission which is measured by a photomultiplier.

The analyzer automatically calculates the cutoff based on the measurement of Cal 1 and Cal2. The result of the samples is given either as reactive or non-reactive as well as in the form of a cutoff index (signal samples/cutoff). Samples with a cutoff index (COI) < 0.7 are non-reactive. Samples with a cutoff index between 0.7 and < 1.0 are considered indeterminate (border). Samples with a cutoff index ≥ 1.0 are considered reactive.

(2) Elecsys PreciControl CMV IgM contains liquid control serum based on human serum. The controls are used for monitoring the accuracy of the Elecsys CMV IgM immunoassay.

The reagents and calibrators are packaged together in the Elecsys CMV IgM assay kit, while the associated Elecsys PreciControl CMV IgM is packaged separately.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study details for the Elecsys CMV IgM Immunoassay, based on the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The document primarily focuses on demonstrating that the device met its predefined acceptance criteria rather than explicitly listing the criteria in a single table with performance. However, I can construct a table summarizing the performance metrics and their implicit acceptance criteria from the "Analytical Performance Data" and "Clinical Performance Data" sections.

Study/Test Area	Acceptance Criteria (Implicit)	Reported Device Performance
Precision (Repeatability)	CV (%) UCL95% values to be within acceptable limits (specific limits not explicitly stated, but implied by passing results).	All repeatability CVs ranged from 0.9% to 2.0% for controls and 1.0% to 1.8% for human serum samples. All met pre-defined acceptance criteria.
Precision (Intermediate)	CV (%) UCL95% values to be within acceptable limits.	All intermediate precision CVs ranged from 2.9% to 5.3% for controls and 2.0% to 4.4% for human serum samples. All met pre-defined acceptance criteria.
Reagent Stability (Unopened)	Stable for 15 months at 2-8°C.	Stable for up to 18 months at 2-8°C, supporting the 15-month claim.
Reagent Stability (Opened)	Stable for 12 weeks at 2-8°C.	Stable for up to 13 weeks at 2-8°C, supporting the 12-week claim.
On-Board Reagent Stability	Stable for 2 weeks at 20°C ± 3°C.	Stable for 34 days at 20°C ± 3°C, supporting the 2-week claim.
On-Board/Refrigerated R. Stability	Stable for 6 weeks when stored alternately in refrigerator and on analyzer.	Met acceptance criteria for 6 weeks.
Sample Stability (2-8°C)	Stable for 4 weeks at 2-8°C.	Stable for up to 35 days (5 weeks) at 2-8°C, supporting the 4-week claim.
Sample Stability (20-25°C)	Stable for 7 days at 20-25°C.	Stable for up to 10 days at 20-25°C, supporting the 7-day claim.
Sample Stability (-20°C)	Stable for 3 months at -20°C.	Stable for approximately 4 months (119 days) at -20°C, supporting the 3-month claim.
Sample Stability (Freeze/Thaw)	Stable through 5 freeze/thaw cycles.	Stable through 5 freeze/thaw cycles, with samples aliquoted for 6 cycles.
Calibration Stability (Lot)	Stable for 28 days.	Met acceptance criteria for 28 days.
Calibration Stability (On-Board)	Stable for 8 days (on the Elecsys 2010 analyzer).	Met acceptance criteria for 8 days.
Calibrator Stability (2-8°C)	Stable for 8 weeks at 2-8°C.	Met acceptance criteria for 8 weeks (tested for 10 weeks).
Open Vial Calibrator Stability	Stable for 5 hours.	Met acceptance criteria for 5 hours.
Control Stability (2-8°C)	Stable for 8 weeks at 2-8°C.	Met acceptance criteria for 8 weeks.
Control Open Vial Stability	Stable for 5 hours.	Met acceptance criteria for 5 hours.
Control Shelf-Life Stability	Stable for 15 months.	Met acceptance criteria for 15 months (tested during and beyond 18 months).
High Dose Hook Effect	No hook effect up to a specified high concentration.	No high dose hook effect observed up to 32.9 COI.
Endogenous & Drug Interferences	For COI < 0.7, non-reactive; for COI ≥ 0.7, recovery ±15%. Rheumatoid factor agreement > 90% against comparator. No interference from 20 pharmaceutical compounds.	Hemoglobin: < 0.621 mmol/L; Bilirubin: < 342 umol/L; Intralipid: < 1500 mg/dL; Biotin: < 410 nmol/L; Rheumatoid factor: < 2000 IU/mL. All results met acceptance criteria for pharmaceutical compounds.
Sample Matrix Equivalence	Acceptable differences in COI recovery relative to serum: <0.07 or between 0.07-0.1 for non-reactive; <10%, 10-20% difference for reactive.	For non-reactive: Li-Heparin Plasma (93% <0.07, 7% 0.07-0.1), K2-EDTA Plasma (100% <0.07), K3-EDTA Plasma (97% <0.07, 3% 0.07-0.1), Serum Separator Tubes (100% <0.07). For reactive: Li-Heparin Plasma (68% <10%, 32% 10-20%), K2-EDTA Plasma (100% <10%), K3-EDTA Plasma (79% <10%, 21% 10-20%), Serum Separator Tubes (90% <10%, 10% 10-20%). All results support use.
Method Comparison (Platform Equivalence)	Positive and negative agreement between platforms demonstrating equivalence.	Negative Percent Agreement (NPA) = 97.98%; Positive Percent Agreement (PPA) = 99.22% between Elecsys 2010 and MODULAR ANALYTICS E170.
Clinical Performance (Prospective Suspected Infection)	High Negative Percent Agreement (NPA) and Positive Percent Agreement (PPA).	NPA: 97.2% (384/395) with a 95% CI of 95.1 – 98.6%. PPA: 87.0% (20/23) with a 95% CI of 66.4 – 97.2%. (Note: 3 equivocal samples from comparator consensus were treated as "not in favor of Elecsys CMV IgM").
Clinical Performance (Preselected Confirmed CMV IgM Positive)	High Positive Percent Agreement (PPA).	PPA: 100.0% (134/134) with a 95% CI of 97.3 – 100.0%.
IgM Specificity	DTT treatment should eliminate or significantly reduce signal, confirming IgM specificity.	10 of 11 samples rendered non-reactive; 1 sample had an 82% decrease in COI. Confirmed specificity for IgM.

2. Sample Sizes and Data Provenance for the Test Set

Clinical Performance Test Set (Method Comparison):
- Prospective Study (Suspected Infection Population): 617 samples.
  - Data Provenance: US, prospective. Samples were from patients suspected of CMV infection for whom a CMV IgM test was ordered. A subset of 199 were pregnant women.
- Retrospective Study (Preselected Confirmed Positive Population): 134 samples.
  - Data Provenance: Retrospective. Samples were preselected from patients positive for CMV IgM as determined by comparator device consensus.
Analytical Performance Test Sets:
- Precision: Not explicitly stated as a general "test set" size, but individual samples/pools were tested (e.g., Controls 01, 02, HS A-F, HS 01-07).
- Cross-Reactivity/Analytical Specificity: 205 specimens.
- High Dose Hook Effect: 5 highly positive, high-titer CMV IgM human samples were diluted.
- Endogenous and Drug Interferences: 8 CMV IgM samples (negative, near cutoff, positive) for hemoglobin, bilirubin, Intralipid, biotin. 34 samples for rheumatoid factor. 20 pharmaceutical compounds.
- Sample Matrix Equivalence: Samples (negative, near cutoff, positive) were collected into matched serum and plasma collection tubes (number not explicitly stated per matrix type).
- Method Comparison Between Analyzer Platforms: Native serum samples (number not specified).
- Reproducibility (Clinical Reproducibility): 4 serum pools and 2 PreciControl CMV IgM materials (n=90 measurements per sample type across sites).

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications

The ground truth for the clinical performance studies was established using comparator FDA-cleared devices, not human experts.

4. Adjudication Method for the Test Set

For the clinical performance method comparison, a two-out-of-three approach was used when three FDA-cleared devices could be tested to determine the consensus result.
When only two FDA-cleared devices could be tested, results for which the devices disagreed were omitted.
Equivocal results from the Elecsys CMV IgM assay were treated as "not in favor of the Elecsys CMV IgM assay" during analysis for the prospective suspected infection population (i.e., if it was equivocal, and the consensus was negative, it was counted against the device).

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

No multi-reader multi-case (MRMC) comparative effectiveness study was mentioned. The device is an in vitro diagnostic (IVD) immunoassay, not an imaging or diagnostic support tool requiring human reader interpretation in the context of MRMC studies.

6. Standalone Performance

Yes, the studies presented are standalone performance of the algorithm (the Elecsys CMV IgM immunoassay) without human-in-the-loop performance. The results are based on the direct output of the immunoassay compared to a comparator consensus or expected values.

7. Type of Ground Truth Used

The ground truth was established by consensus results from multiple (2 or 3) FDA-cleared comparator CMV IgM assays. This is a form of "reference method" ground truth.

8. Sample Size for the Training Set

The document does not explicitly mention a separate "training set" in the context of machine learning. For traditional IVDs like this immunoassay, calibration and assay optimization would be part of the development process.

Assay Cut-off Establishment:
- Initial cutoff establishment: Samples characterized using several commercially available CMV IgG and CMV IgM assays (number not specified).
- Specificity evaluation: 931 samples from a low prevalence cohort.
- Sensitivity optimization: 152 samples from individuals at different stages of primary CMV infection.
- Validation of assay cutoff: Performed by external clinical studies (the test set described in #2).

9. How the Ground Truth for the Training Set was Established

As noted in #8, there isn't a "training set" in the common machine learning sense. For the development and optimization of the assay:

The "ground truth" for developing and optimizing the assay's cutoff was derived from characterizing samples using several commercially available CMV IgG and CMV IgM assays and by using samples from individuals at different stages of primary CMV infection. These are established laboratory methods and clinical diagnoses typically relying on a combination of existing cleared tests and clinical context.

Summary

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

October 24, 2014

ROCHE DIAGNOSTICS SARAH BAUMANN PROGRAM MANAGER, REGULATORY AFFAIRS 9155 HAGUE RD. INDIANAPOLIS, IN 46250

Re: K142133

Trade/Device Name: Elecsys CMV IgM Immunoassay: Elecsys PreciControl CMV IgM Regulation Number: 21 CFR 866.3175 Regulation Name: Cytomegalovirus serological reagents Regulatory Class: II Product Code: LFZ, JJX Dated: August 1, 2014 Received: August 4, 2014

Dear Ms. Baumann:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours.

Stephen J. Lovell -S for

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K142133

Device Name

(1) Elecsys CMV IgM Immunoassay

(2) Elecsys PreciControl CMV IgM

Indications for Use (Describe)

The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys immunoassay analyzers.

(2) PreciControl CMV IgM is used for quality control of the Elecsys CMV IgM immunoassay on the Elecsys and cobas e immunoassay analyzers.

Type of Use (Select one or both, as applicable)

2 Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

FOR FDA USE ONLY

Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

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1 Section III—510(k) Summary

1.1 Introduction

According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

1.2 Submitter Name, Address, Contact Information

Roche Diagnostics Corporation 9115 Hague Road Indianapolis, IN 46250

	Contact Person:	Sarah Baumann, MS, RAC
	Phone:	317-521-3952
	Fax:	317-521-2324
	Email:	sarah.baumann@roche.com

	Secondary Contact Person:	Dr. Michael Leuther, Ph.D.
	Phone:	317-521-3930
	Fax:	317-521-2324
	Email:	michael.leuther@roche.com

Date Prepared: October 23, 2014

1.3 Device Name

Proprietary Name:	(1) Elecsys CMV IgM Immunoassay(2) Elecsys PreciControl CMV IgM
Common Name:	(1) CMV IgM assay(2) PreciControl CMV IgM
Classification Name:	(1) Enzyme Linked Immunoabsorbent Assay,Cytomegalovirus(2) Single (Specified) Analyte Controls (Assayed andUnassayed)
Product Code:	(1) LFZ(2) JJX
Predicate Device:	(1) Diamedix Is-CMV IgM Capture Test Kit (K001767)(2) Elecsys PreciControl Rubella IgM (K092322)

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Panel	ProductCode	Classification Name	RegulationCitation
		Enzyme Linked
Microbiology	LFZ	Immunoabsorbent Assay,	21 CFR 866.3175
		Cytomegalovirus
	JJX	Single (Specified)
Clinical Chemistry		Analyte Controls	21 CFR 862.1660
		(Assayed and Unassayed)

1.4 Device Description

(1) Elecsys CMV IgM:

Elecsys CMV IgM is a u-capture immunoassay with streptavidin microparticles, biotinylated recombinant CMV-specific antigen labeled with a ruthenium complex and electrochemiluminescence detection. The results are determined using a calibration curve which is instrument-specifically generated by a 2-point calibration and a master curve provided via the reagent bar code. The test system contains the human serum-based calibrators intended for use with the system.

The CMV IgM assay begins with an automatic 1:20 predilution of sample with Elecsys Diluent Universal and the addition of biotinylated monoclonal anti-h-IgM-specific antibodies.

(2) Elecsys PreciControl CMV IgM:

Elecsys PreciControl CMV IgM contains liquid control serum based on human serum. The controls are used for monitoring the accuracy of the Elecsys CMV IgM immunoassay.

The reagents and calibrators are packaged together in the Elecsys CMV IgM assay kit, while the associated Elecsys PreciControl CMV IgM is packaged separately.

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The following reagents are provided in the Elecsys CMV IgM assay kit:

1. The reagent rackpack consists of reagents M, R1, and R2 and is labeled as CMVIGM: -M: Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL: Streptavidin-coated microparticles 0.72 mg/mL; preservative.
  -R1: Anti-h-IgM-Ab~biotin (gray cap), 1 bottle, 9 mL: Biotinylated monoclonal anti-h-IgM antibody (mouse) > 500 µg/L, MES buffer 50 mmol/L, pH 6.5; preservative.

-R2: CMV-Ag~Ru(bpy) (black cap), 1 bottle, 9 mL: CMV-specific antigen (recombinant, E. coli) labeled with ruthenium complex > 50 µg/L; MES buffer 50 mmol/L, pH 5.5; preservative.

1. CMVIGM Call: Negative calibrator 1 (white cap), 2 bottles of 1.0 mL each: Human serum, non-reactive for anti-CMV IgM; preservative.
1. CMVIGM Cal2: Positive calibrator 2 (black cap), 2 bottles of 1.0 mL each: Human serum reactive for anti-CMV IgM; buffer; bovine albumin; preservative.

The following are the materials that are required but not provided:

1. PreciControl CMV IgM, 8 x 1.0 mL each of PreciControl CMV IgM 1 and 2
1. Diluent Universal, 2 x 36 mL sample diluent
1. CalSet Vials, 2 x 56 empty snap-cap bottles
1. General laboratory equipment
1. Elecsys 2010 analyzer
1. Accessories for Elecsys 2010 analyzer

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1.5 Intended Use

(1) Elecsys CMV IgM:

Immunoassay for the in vitro qualitative detection of IgM antibodies to CMV in human serum, lithium-heparin plasma, K2-EDTA plasma, and K2-EDTA plasma. The test is intended as an aid in the diagnosis of recent or current CMV infection in indiviuals for which a CMV IgM test was ordered, including pregnant women.

Performance characteristics have not been evaluated in immunocompromised or immunosuppressed individuals. This test is not intended for use in neonatal screening or for use at point of care facilities. This assay is not intended for use in screening blood and plasma donors.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys immunoassay analyzers.

(2) Elecsys PreciControl CMV IgM:

PreciControl CMV IgM is used for quality control of the Elecsys CMV IgM immunoassay on the Elecsys and cobas e immunoassay analyzers.

1.6 Indication for Use

Refer to Section 1.5 (Intended Use) above.

1.7 Substantial Equivalence Information

1.7.1 Predicate Device

The Elecsys CMV IgM Assay is substantially equivalent to other devices legally marketed in the United States and in commercial distribution for the detection of IgM antibodies to cytomegalovirus (CMV) in human serum and plasma.

We claim equivalency to the Diamedix Is-CMV Capture Test Kit (K001767) for the in vitro qualitative detection of IgM antibodies to cytomegalovirus (CMV) in human serum and plasma.

The Elecsys PreciControl CMV IgM quality control materials are substantially equivalent to the Roche Elecsys PreciControl Rubella IgM quality control materials cleared in K092322.

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Comparison with Predicates: Similarities 1.7.2

Table 1 below provides a comparison of the Elecsys CMV IgM assay and the chosen predicate device, the Diamedix Is-CMV IgM Capture Test Kit (K001767).

Feature	Candidate DeviceElecsys CMV IgM Assay	Predicate DeviceDiamedix Is-CMV IgM Capture Test Kit(K001767)
Intended Use/Indications forUse	Immunoassay for the in vitro qualitativedetection of IgM antibodies to CMV in humanserum, lithium-heparin plasma, K2-EDTAplasma, and K3-EDTA plasma. The test isintended as an aid in the diagnosis of recent orcurrent CMV infection in individuals for whicha CMV IgM test was ordered, includingpregnant women.Performance characteristics have not beenevaluated in immunocompromised orimmunosuppressed individuals. This test is notintended for use in neonatal screening or foruse at point of care facilities. This assay is notintended for use in screening blood and plasmadonors.The electrochemiluminescence immunoassay"ECLIA" is intended for use on Elecsysimmunoassay analyzers.	The Diamedix Is-CMV IgM Capture Test Kit is acapture enzyme immunoassay (EIA) for thequalitative detection of IgM antibodies to CMV inhuman serum as an aid in the diagnosis of recentor current infection with CMV. These reagentscan be used either manually or in conjunction withthe MAGO Plus Automated EIA Processor. Thisproduct has not been cleared/approved by theFDA for blood/plasma donor screening.
Assay Protocol	μ-Capture	Solid-phase microtiter
DetectionProtocol	Electrochemiluminescent Immunoassay	Enzyme-linked immuno-adsorbent assay

Table 1. Comparison of Candidate (Elecsys CMV IgM Assay) and Predicate Device

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Analytical Performance Data 1.8

1.8.1 Precision

The following precision results were obtained with the Elecsys CMV IgM assay in human serum samples and quality control materials. Within-run precision (repeatability) and total imprecision (intermediate precision) were determined according to CLSI Guideline EP5-A2. All results met the pre-defined acceptance criteria for repeatability and intermediate precision.

		Repeatability		Intermediate Precision
Sample	Mean (COI)	SD	CV (%)(UCL95%)	SD	CV (%)(UCL95%)
Control 01	0.171	0.003	1.8 (2.9)	0.005	2.9 (4.4)
Control 01	0.220	0.002	0.9 (1.7)	0.007	3.2 (4.6)
Control 02	1.98	0.040	2.0 (2.4)	0.105	5.3 (6.7)
Control 02	2.03	0.032	1.6 (2.1)	0.085	4.2 (5.6)
HS B	0.175	0.003	1.7 (2.8)	0.004	2.3 (3.9)
HS A	0.200	0.002	1.0 (1.7)	0.004	2.0 (3.7)
HS 07	0.249	0.004	1.6 (2.4)	0.007	2.8 (3.7)
HS 04	0.841	0.010	1.2 (1.6)	0.026	3.1 (4.1)
HS 05	0.913	0.013	1.4 (1.8)	0.027	3.0 (4.0)
HS 01	0.957	0.010	1.0 (1.2)	0.031	3.2 (4.4)
HS 06	1.13	0.018	1.6 (2.1)	0.040	3.5 (4.6)
HS 02	1.21	0.016	1.3 (1.7)	0.042	3.5 (4.8)
HS D	1.22	0.022	1.8 (2.2)	0.038	3.1 (3.7)
HS C	1.62	0.024	1.5 (1.8)	0.055	3.4 (4.3)
HS F	4.16	0.067	1.6 (1.9)	0.141	3.4 (4.3)
HS 03	5.07	0.066	1.3 (1.6)	0.157	3.1 (4.1)
HS E	13.9	0.236	1.7 (2.1)	0.445	3.2 (3.9)

Table 2. Summary of Precision Results for Elecsys CMV IgM

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1.8.2 Reagent, Sample, Calibration, Calibrator, and Control Stability

Reagent Stability:

Four studies were conducted to evaluate the stability of the Elecsys CMV IgM assay reagent: reagent stability (unopened) at 2 - 8°C. reagent stability (opened) at 2 - 8°C. onboard reagent stability, and reagent stability stored alternately on-board the analyzer and refrigerated.

1. Reagent Stability (Unopened) at 2 8°C: Shelf-life reagent stability for the Elecsys CMV IgM assay was tested on one Elecsys 2010 immunoassay analyzer. Three lots of reagent were evaluated using PreciControl CMV IgM (Levels 1 and 2). A fresh kit was placed on the analyzer and calibrated. Reference values for the PreciControl CMV IgM Levels 1 and 2 were then determined. After measurement, the kit was closed and stored refrigerated (2-8°C). The on-test reagent was stored at 2-8°C and PC CMV IgM materials tested in duplicate at several time points and the acceptance criteria were met at each time point. The Elecsys CMV IgM reagent was stable unopened at 2 - 8°C up to 18 months, which supports the claimed shelf life of 15 months.
1. Reagent Stability (Opened) at 2 8°C: Reagent stability after first opening for the Elecsys CMV IgM assay was tested on an Elecsys 2010 immunoassay analyzer with duplicate measurements. A fresh kit was placed on the analyzer and calibrated. Reference values for samples on-test were then determined. After measurement, the kit was removed from the analyzer and kept at 2-8°C for up to 13 weeks (92 days). Calibration was repeated and all samples were placed on the analyzer and remeasured. The acceptance criteria were met for reagent stability after opening at 2 - 8℃ for 13 weeks, supporting the claim of 12 weeks.
1. On-Board Reagent Stability: On-board reagent stability for the Elecsys CMV IgM assay was tested on one Elecsys 2010 immunoassay analyzer. A fresh kit was placed on the analyzer and calibrated. Reference values for the samples tested were then determined. After measurement, the kits were closed and stored at 20°C ± 3°C to simulate on-board conditions. At specific time points, the kits were placed on the analyzer, calibrated, and the original test samples re-measured. Samples were measured in duplicate with one reagent lot in one run per day. Acceptance criteria were met at each time point for 34 days, supporting the on-board reagent stability claim of 2 weeks.
1. On-Board/Refrigerated Reagent Stability: On-board/refrigerated reagent stability for the Elecsys CMV IgM assay was tested on one Elecsys 2010 immunoassay analyzer by comparing the stressed samples with the reference samples. A fresh kit was placed on the analyzer and calibrated. Reference values for the samples tested were then determined. After measurement, the kit was closed and kept 5x4 hours on-board (20°C ± 3°C) per week; the rest of the time it was stored refrigerated (2-8°C). The acceptance criteria were met, supporting reagent stability up to 6 weeks when stored alternately in the refrigerator and on the analyzer.

Sample Stability:

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Four studies were conducted to evaluate sample stability: sample stability at 2 - 8°C. sample stability at 20 - 25℃, sample stability at -20℃, and sample stability through multiple freeze/thaw cycles.

1. Sample Stability at 2 8°C: Reference and on-test materials were tested in triplicate on the Elecsys 2010 analyzer. Samples were measured directly after collection (reference) and then stored at 2-8°C for 35 days. These samples were tested after storage at 2-8°C at multiple time points up to 35 days. The on-test recovery was calculated as a percent of the reference value. The acceptance criteria were met, supporting sample stability up to 4 weeks when stored at 2 - 8°C.
1. Sample Stability at 20 - 25°C: Reference and on-test material (human serum and plasma samples) was tested in triplicate and the controls were tested in duplicate on the Elecsys 2010 analyzer. The samples were measured directly after collection (reference) and then stored at 20-25°C for up to 10 days. The on-test recovery was calculated as a percent of or difference from the reference value. The acceptance criteria were met, supporting claimed sample stability up to 7 days at 20 - 25°C,
1. Sample Stability at -20°C: Reference and on-test material was tested in triplicate on the Elecsys 2010 analyzer. The samples were measured immediately after collection (reference) and then stored frozen at -20°C for approximately 4 months (119 days). The samples were re-measured after 119 days of frozen storage at -20°C. The ontest recovery was calculated as a percent of or difference from the reference value. The acceptance criteria were met, supporting claimed sample stability for 3 months at -20°C.
1. Sample Stability Through Multiple Freeze/Thaw Cycles: Reference and on-test material was tested in triplicate on the Elecsys 2010 analyzer. The samples were measured immediately after collection (reference) and aliquots were frozen/thawed 6 times. A sample measurement was taken after each freeze/thaw cycle and the results compared to the reference result. The acceptance criteria were met, supporting claimed sample stability through 5 freeze/thaw cycles,

Calibration Stability:

Two studies were conducted to evaluate calibration stability: lot calibration stability and on-board calibration stability.

1. Lot Calibration Stability: The lot calibration stability was determined by comparing the calibration for two kits of the same lot. On Day 1, the first reagent kit was opened and calibrated, and samples were measured at Days 1 (reference), 29 and 36 on one Elecsys 2010 analyzer. All samples were measured in duplicate with one reagent in one run per day on Days 1, 29, and 36. The acceptance criteria were met for lot calibration stability of 28 days on the Elecsys 2010 analyzer.
1. On-Board Calibration Stability: On-board calibration stability for the Elecsys CMV IgM test system was tested on one Elecsys 2010 analyzer. One reagent kit was

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opened and samples were measured on Day 1 (reference). The same samples were then retested after 15 days with reagent bottles stored at 20 ± 3℃ (on-board condition) using the calibration from Day 1. Recovery was calculated based on the initial (reference) values. All samples were tested in duplicate with one reagent lot on one Elecsys 2010 analyzer in one run per day on two days. The acceptance criteria were met for on-board calibration stability of 8 days on the Elecsys 2010 analyzer.

Calibrator Stability:

Two studies were conducted to evaluate calibrator stability: calibrator stability at 2 - 8°C and open vial calibrator stability.

1. Calibrator Stability at 2 8°C: Reference and on-test materials were tested in duplicate with one reagent lot in one run per day on one Elecsys 2010 analyzer. The on-test material was opened and stored at 2 - 8°C for 8 weeks. It was measured at Day 1 (reference) and again at 8 and 10 weeks. The on-test recovery was calculated as the signal (counts) of the reference value. The acceptance criteria were met for calibrator stability at 2 - 8℃ for 8 weeks.
1. Open Vial Calibrator Stability: Reference and open vial, on-test materials were tested in duplicate with one reagent lot on one Elecsys 2010 analyzer. The on-test material was opened and stored at 25°C for 6 hours to simulate on-board conditions. Open vial (on-test) material was also stored at 32°C for 6 hours on one Elecsys 2010 to simulate on-board conditions. Every hour, each calibrator was tested and the on-test recovery of signal (counts) was calculated as a percent of the reference value. The acceptance criteria were met for open vial calibrator stability for 5 hours on the Elecsys 2010 analyzer.

Preci-Control Value Assignment:

The Elecsys CMV IgM calibrators and Elecsys PreciControl CMV IgM values are assigned through at least six independent series of analysis performed on at least three analyzers. All samples are tested in duplicate. The sample recovery (COI) is calculated as the median of each sample as reference to the target value.

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Stability of PreciControl CMV IgM:

Three studies were conducted to evaluate the stability of the Elecsys PreciControl CMV IgM materials: control stability at 2 - 8°C, open vial control stability, and shelf-life stability.

1. Control Stability at 2 8℃: Stressed and unstressed samples of PreciControl CMV IgM were tested in duplicate with one reagent lot in one run per day on one Elecsys 2010 analyzer. All samples met the acceptance criteria, supporting the claimed PreciControl CMV IgM stability for 8 weeks when stored at 2 - 8°C.
1. Control Open Vial Stability: Reference and open vial, on-test controls were tested in singlicate with one reagent lot on one Elecsys 2010 analyzer. The on-test material was opened, kept at 25℃ on the Elecsys 2010 to simulate on-board conditions, and tested every hour for 6 hours. The on-test recovery of signal (counts) was calculated as a percent of the reference value. The acceptance criteria were met, supporting the claimed PreciControl CMV IgM open vial stability of 5 hours on the Elecsys 2010 analyzer.
1. Control Shelf-Life Stability: The on-test material was stored at 2-8°C and tested in duplicate on the Elecsys 2010 analyzer at several time points during and beyond the shelf life time of 18 months. Stability was measured by comparing the measurements of the stressed controls with the measurements of the unstressed controls (reference). The acceptance criteria were met at each time point, supporting the claimed PreciControl CMV IgM shelf-life stability of 15 months.

1.8.3 High Dose Hook Effect

Testing with the Elecsys CMV IgM assay demonstrated no high dose hook effect. Five highly positive. high-titer CMV IgM human samples were diluted with negative human serum in a dilution series with 11 steps. Each dilution was tested in triplicate with one reagent lot in one run on one Elecsys 2010 analyzer. No high dose hook effect was observed up to 32.9 COI.

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1.8.4 Cross-Reactivity/Analytical Specificity

A study was conducted to evaluate the Elecsys CMV IgM for potential cross-reactivity using samples from individuals with antibodies to various medical conditions. Specimens (n = 205) were tested in duplicate with the Elecsys CMV IgM assay and a comparator CMV IgM assay. The first result was used for the performance evaluation. The presence of the potential cross-reactants in the samples tested was confirmed using FDA-cleared devices or laboratory methods. Potential cross-reactivity with autoimmune markers and antibodies against influenza vaccination could not be ruled out from the study.

PotentialCross-	Comparator CMV IgM Results(Negative)			Comparator CMV IgM Results(Positive)
Reactingantibodiesand diseaseconditions	ElecsysCMVIgMNon-Reactive	Elecsys CMVIgMIndeterminate(Border)	ElecsysCMVIgMReactive	ElecsysCMVIgMNon-Reactive	Elecsys CMVIgMIndeterminate(Border)	ElecsysCMVIgMReactive
Autoimmune	5	0	2	0	0	0
EBV	14	0	0	0	0	0
E. coli	5	0	0	0	0	0
HCV	27	0	0	0	0	0
HSV	4	0	0	1	0	0
HTLV	43	0	0	0	0	0
Rubella	10	0	0	0	0	0
HAV	10	0	0	0	0	0
HBV	18	0	0	1	0	0
HIV	20	0	0	1	0	0
InfluenzaVaccine	9	0	1	0	0	0
Treponemapalladium	4	0	0	0	0	0
Toxoplasmagondii	9	0	0	0	0	0

Table 3. Cross-Reactivity Test Results

1.8.5 Endogenous and Drug Interferences

To evaluate the effect of elevated levels of hemoglobin, bilirubin, Intralipid, and biotin on the CMV IgM assay, eight CMV IgM samples (negative, near cutoff and positive) were spiked with the potential interferents. Each interferent was evaluated at 11 numerical values. All samples were tested in singlicate on the Elecsys 2010 analyzer. Acceptance criteria for hemoglobin, bilirubin, Intralipid, and biotin: samples with COI < 0.7 to be found non-reactive, and samples with COI ≥ 0.7 recovery of ±15%.

To evaluate the effect of rheumatoid factor, 34 samples were spiked with different concentrations of RF and measured in singlicate on the Elecsys 2010 analyzer as well as

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a comparator device. The acceptance criterion for rheumatoid factor was agreement > 90% against the comparator device.

Interferent tested	No interference up to (Concentration)
Hemoglobin	< 0.621 mmol/L or < 1.0 g/dL
Bilirubin	< 342 umol/L or < 20 mg/dL
Intralipid	< 1500 mg/dL
Biotin	< 410 nmol/L or < 100 ng/mL
Rheumatoid factor	< 2000 IU/mL

The results of the interference studies are presented below:

In addition, 20 pharmaceutical compounds were evaluated for interference with the Elecsys CMV IgM assay. All results met the pre-defined acceptance criteria, demonstrating no interference from the drug substances tested.

Studies were conducted to evaluate the suitability of the following types of samples to be used with the Elecsys CMV IgM immunoassay: Li-heparin plasma, K2-EDTA plasma, K3-EDTA plasma, and serum separator tubes. Samples were collected into matched serum and plasma collection tubes and assayed in duplicate. These studies were conducted using negative (non-reactive), near cutoff (indeterminate/border) and positive (reactive) samples. The results support the use of the following sample types: Li-heparin plasma, K2-EDTA plasma, K3-EDTA plasma, and serum separator tubes. Acceptance specifications are tabulated below.

Sample Matrix	Mean COI	Percent of samples showing differences in recovery relative to serum (COI) in non-reactive specimens
		< 0.07 COI	0.07 – 0.1 COI	> 0.1 COI
Li-Heparin Plasma	0.196	93 %	7 %	0 %
K2-EDTA Plasma	0.280	100 %	0 %	0 %
K3-EDTA Plasma	0.195	97 %	3 %	0 %
Serum Separator Tubes	0.265	100 %	0 %	0 %

Sample Matrix	Mean COI	Percent of samples showing differences inrecovery relative to serum (COI) inreactive specimens
		< 10 %	10 - 20 %	> 20 %
Li-Heparin Plasma	1.51	68 %	32 %	0 %
K2-EDTA Plasma	2.12	100 %	0 %	0 %
K3-EDTA Plasma	1.72	79 %	21 %	0 %
Serum Separator Tubes	2.27	90 %	10 %	0 %

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1.8.6 Method Comparison Between Analyzer Platforms

The equivalence of the Elecsys CMV IgM assay on the Elecsys 2010 and MODULAR ANALYTICS E170 immunoassay analyzers was evaluated by a method comparison study. Native serum samples were tested on one Elecsys 2010 and one MODULAR ANALYTICS E170 analyzer. Positive and negative agreement of the results between the two platforms was calculated and demonstrated equivalence between the analyzer platforms for the determination of IgM antibodies to CMV using the Elecsys CMV IgM assay:

Concordance Rates:

Negative Percent Agreement (NPA) = 97.98 % Positive Percent Agreement (PPA) = 99.22 %

1.8.7 Assay Cut-Off

The cut-off for the Elecsys CMV IgM assay was established with in-house studies by characterizing samples using several commercially available CMV IgG and CMV IgM assays. Specificity of the assay was evaluated using 931 samples from a low prevalence cohort. The assay cutoff was optimized to increase sensitivity by measuring 152 samples from individuals at different stages of primary CMV infection. Validation of the assay cutoff was performed by external clinical studies.

1.8.8 Verification of IgM Specificity

To confirm that the Elecsys CMV IgM assay specifically detects IgM-class antibodies, 11 samples with moderate to high levels of CMV IgM antibodies were selected for testing. These samples were treated with dithiothreitol (DTT) to destroy the IgM and then were retested using the Elecsys CMV IgM assay. Ten samples were rendered non-reactive (negative) following treatment with DTT, and one sample had an 82% decrease in COI value, confirming the specificity of the Elecsys CMV IgM assay for detecting IgM-class antibodies. The Elecsys CMV IgM assay is specific for anti-CMV IgM antibodies due to the u-capture design and antigens minimizing interference from CMV IgG antibodies.

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Clinical Performance Data 1.9

Reproducibility 1.9.1

The reproducibility was assessed by testing the Elecsys CMV IgM assay on the Elecsys 2010 analyzer at three sites. Imprecision testing was conducted using three replicates in two runs per day for five days consistent with requirements within CLSI EP5-A2 and EP15-A2 with four serum pools and two PreciControl CMV IgM materials. One reagent lot was used for all of the testing at all three sites. The sample panels evaluated included samples close to the cutoff, reactive and non-reactive. All sample panels demonstrated a CV % less than 10 % and the results are presented in the table below.

Table 4. Elecsys CMV IgM Clinical Reproducibility

		Repeatability		Between-Day		Between-Site		Reproducibility
Sample	n	MeanCOIa	SDb	% CV	SD	% CV	SD	% CV	SD	% CV
HSP 01 Near Cut-Off	90	0.839	0.012	1.4	0.000c	0.0	0.039	4.6	0.043	5.1
HSP 02 Near Cut-Off	90	1.094	0.016	1.5	0.000c	0.0	0.047	4.3	0.052	4.8
HSP 03 Reactive	90	5.156	0.126	2.4	0.027	0.5	0.058	1.1	0.186	3.6
HSP 04 Non-Reactive	90	0.242	0.003	1.3	0.002	0.7	0.023	9.6	0.024	9.8
PC CMV-IgM_1 Negative	90	0.207	0.003	1.6	0.003	1.4	0.019	9.2	0.020	9.5
PC CMV-IgM_2 Positive	90	1.851	0.042	2.3	0.000c	0.0	0.042	2.3	0.079	4.2
a COI - Cutoff indexb SD - Standard deviationc SD of zero due to variance contributed by particular component was below stated significant figure.

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1.9.2 Method Comparison

A multi-center study was conducted in the US to evaluate the performance of the Elecsys CMV IgM immunoassay in comparison to a consensus result derived from 2 or 3 FDA-cleared devices (depending on the availability of sample volume for testing). A two-out-of-three approach was used to determine the consensus result for samples where three FDA-cleared devices could be tested. Where only two FDA-cleared devices could be tested, results for which the FDA-cleared devices disagreed were omitted from analysis. All equivocal results obtained using the Elecsys CMV IgM assay were treated as not in favor of the Elecsys CMV IgM assay.

The main study was a prospective study consisting of 617 samples, all of which were from a population of patients suspected of CMV infection for whom a CMV IgM test was ordered. A subset of this population (n = 199) were pregnant women. In addition, a retrospective study was conducted, consisting of 134 samples that were preselected from a population of patients positive for CMV IgM as determined by the comparator device consensus of 3 FDA-cleared CMV IgM assays. Testing of specimens was done at three clinical sites and one internal site. The results of these studies are presented in the following tables.

		Comparator Device ConsensusCMV IgM Result
		Positive	Equivocal	Negative	Total
ElecsysCMVIgMResult	Reactive	20	0	6	26
	Indeterminate(Border)	0	0	5	5
	Non-Reactive	0	3a	384	387
	Total	20	3	395	418
AgreementClassification	Numerator/Denominator	Percent Agreement (%)		95% ConfidenceInterval
Negative Agreement	384/395	97.2		95.1 – 98.6
Positive Agreement	20/23	87.0		66.4 – 97.2

Table 5. Prospectively Collected Suspected Infection Population

a These samples were Equivocal according to the consensus result and Non-reactive (negative) by the Elecsys CMV IgM assay. As such, they were treated as Elecsys CMV IgM Reactive (positive) in the analysis (i.e., not in favor of the Elecsys CMV IgM assay).

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	Comparator Device ConsensusCMV IgM Result
		Positive	Equivocal	Negative	Total
ElecsysCMVIgMResult	Reactive	0	0	2	2
	Indeterminate(Border)	1	0	1	2
	Non-Reactive	0	5	190	195
	Total	1	5	193	199

Table 6. Prospectively Collected Pregnant Population

Table 7. Preselected Confirmed CMV IgM Positive Population

		Comparator Device Consensus
		CMV IgM Result
		Positive	Equivocal	Negative	Total
ElecsysCMVIgMResult	Reactive	134	0	0	134
	Indeterminate(Border)	0	0	0	0
	Non-Reactive	0	0	0	0
	Total	134	0	0	134

AgreementClassification	Numerator/Denominator	PercentAgreement (%)	95% ConfidenceInterval
Negative Agreement		N/Ab
Positive Agreement	134/134	100.0	97.3 – 100.0

ኮ The number of negative samples in this study was not sufficient (and not required) to make statistically valid conclusions about the negative percent agreement.

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Expected Values/Reference Range 1.9.3

The observed expected values in the prospectively collected US clinical study population using the Elecsys CMV IgM assay were as follows:

Age Group(years)	Sex	ReactiveN (%)	Indeterminate (Border)N (%)	Non-ReactiveN (%)	Total (N)
18 – 19	Female	0 (0.00)	0 (0.00)	2 (100.00)	2
	Male	0 (0.00)	0 (0.00)	5 (100.00)	5
20 - 29	Female	2 (1.96)	1 (0.98)	99 (97.06)	102
	Male	0 (0.00)	2 (4.44)	43 (95.56)	45
30 - 39	Female	2 (1.23)	2 (1.23)	159 (97.55)	163
	Male	1 (1.30)	2 (2.60)	74 (96.10)	77
40 - 49	Female	1 (3.03)	0 (0.00)	32 (96.97)	33
	Male	0 (0.00)	0 (0.00)	70 (100.00)	70
50 - 59	Female	2 (12.50)	0 (0.00)	14 (87.50)	16
	Male	1 (1.67)	0 (0.00)	59 (98.33)	60
60 – 69	Female	0 (0.00)	0 (0.00)	6 (100.00)	6
	Male	1 (5.56)	0 (0.00)	17 (94.44)	18
70 - 79	Male	0 (0.00)	0 (0.00)	2 (100.00)	2
Totals	Female	7 (2.17)	3 (0.93)	312 (96.89)	322
	Male	3 (1.08)	4 (1.44)	270 (97.47)	277
	Overall	10 (1.67)	7 (1.17)	582 (97.16)	599

Table 8. Expected Values—Elecsys CMV IgM Assay

Regulation Number and Section

§ 866.3175 Cytomegalovirus serological reagents.

(a)
Identification. Cytomegalovirus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to cytomegalovirus in serum. The identification aids in the diagnosis of diseases caused by cytomegaloviruses (principally cytomegalic inclusion disease) and provides epidemiological information on these diseases. Cytomegalic inclusion disease is a generalized infection of infants and is caused by intrauterine or early postnatal infection with the virus. The disease may cause severe congenital abnormalities, such as microcephaly (abnormal smallness of the head), motor disability, and mental retardation. Cytomegalovirus infection has also been associated with acquired hemolytic anemia, acute and chronic hepatitis, and an infectious mononucleosis-like syndrome.(b)
Classification. Class II (performance standards).