DIASORIN LIAISON EBNA IGG, LIAISON VCA IGG, LIAISON VCA IGM ASSAYS

K040120 · DiaSorin, Inc. · LLM · Apr 29, 2005 · Microbiology

Device Facts

• Record ID: K040120
• Device Name: DIASORIN LIAISON EBNA IGG, LIAISON VCA IGG, LIAISON VCA IGM ASSAYS
• Applicant: DiaSorin, Inc.
• Product Code: LLM · Microbiology
• Decision Date: Apr 29, 2005
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3235
• Device Class: Class 1

Intended Use

The LIAISON® VCA IgG assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgG antibodies to Epstein-Barr virus (EBV) viral capsid antigen (VCA) p18 synthetic peptide in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis. The LIAISON® EBV IgM assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgM antibodies to Epstein-Barr virus (EBV) viral capsid antigen (VCA) p18 synthetic peptide in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis. The LIAISON® EBNA IgG assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgG antibodies to Epstein-Barr virus (EBV) nuclear antigen synthetic peptide (EBNA-1) in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis.

Device Story

The LIAISON® EBNA IgG, EBV IgM, and VCA IgG assays are chemiluminescence immunoassays (CLIA) performed on the LIAISON® Analyzer. Input: human serum samples. Principle: indirect CLIA using magnetic particles coated with synthetic peptides (EBNA-1 or VCA p-18) and isoluminol-antibody conjugates. The analyzer performs incubation, wash cycles, and starter reagent addition; a photomultiplier measures light signal (RLU) proportional to antibody concentration. Used in clinical laboratories by technicians. Output: qualitative detection of specific IgG or IgM antibodies. Results aid clinicians in diagnosing EBV-associated syndromes and determining immune status. Benefits: provides standardized, automated serological testing for EBV infection.

Clinical Evidence

Clinical trials conducted at two US labs and DiaSorin using prospective and retrospective serum samples. Prospective sample agreement: EBNA IgG (95.3%), EBV IgM (89.4%), VCA IgG (96.1%). Retrospective (VCA IgM-positive) agreement: EBNA IgG (94.3%), EBV IgM (95.7%), VCA IgG (100%). Studies included reproducibility testing across sites and cross-reactivity panels for herpes viruses and other conditions. No conclusive evidence of interference observed.

Technological Characteristics

Indirect chemiluminescence immunoassay (CLIA). Solid phase: magnetic particles coated with synthetic EBV peptides. Conjugate: mouse monoclonal antibody to human IgG/IgM linked to isoluminol derivative. Detection: flash chemiluminescence measured as RLU via photomultiplier. Automated on LIAISON® Analyzer. Qualitative output.

Indications for Use

Indicated for the laboratory diagnosis of EBV-associated infectious mononucleosis in individuals with signs and symptoms consistent with infectious mononucleosis.

Regulatory Classification

Identification

Epstein-Barr virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to Epstein-Barr virus in serum. The identification aids in the diagnosis of Epstein-Barr virus infections and provides epidemiological information on diseases caused by these viruses. Epstein-Barr viruses are thought to cause infectious mononucleosis and have been associated with Burkitt's lymphoma (a tumor of the jaw in African children and young adults) and postnasal carcinoma (cancer).

Predicate Devices

• DiaSorin ETI-VCA-G Kit (K946159)
• DiaSorin ETI-EBV-M reverse Kit (K946157)
• DiaSorin ETI-EBNA-G Kit (K946158)

Submission Summary (Full Text)

{0}

510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
DEVICE ONLY TEMPLATE

A. 510(k) Number:
k040120

B. Purpose for Submission:
New device clearance

C. Measurand:
Epstein Barr Virus IgG to Viral Capsid Antigen, Epstein Barr Virus IgM to Viral
Capsid Antigen, and Epstein Barr Virus IgG to Nuclear Antigen

D. Type of Test:
Qualitative, CLIA

E. Applicant:
DiaSorin Inc.

F. Proprietary and Established Names:
DiaSorin LIAISON® VCA IgG
DiaSorin LIAISON® VCA IgM
DiaSorin LIAISON® EBNA IgG

G. Regulatory Information:
a) Regulation section:
Epstein-Barr Virus, serological reagents (21 CFR 866.3235).
b) Classification:
Class I
Product Code:
LSE
LLM
c) Panel:
83 Microbiology

H. Intended Use:
a) Intended use(s):
The LIAISON® VCA IgG assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgG antibodies to Epstein-Barr virus (EBV) viral capsid antigen (VCA) p18 synthetic peptide in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis.

{1}

Page 2 of 14

The LIAISON® EBV IgM assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgM antibodies to Epstein-Barr virus (EBV) viral capsid antigen (VCA) p18 synthetic peptide in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis.

The LIAISON® EBNA IgG assay uses chemiluminescent immunoassay (CLIA) technology on the LIAISON® Analyzer for the qualitative determination of specific IgG antibodies to Epstein-Barr virus (EBV) nuclear antigen synthetic peptide (EBNA-1) in human serum. When performed in conjunction with other EBV markers, this assay can be used as an aid in the clinical laboratory diagnosis of Epstein-Barr Viral Syndrome in patients with signs and symptoms of EBV infection such as infectious mononucleosis.

b) Indication(s) for use:
The LIAISON® VCA IgG, the LIAISON® VCA IgM, and the LIAISON® EBNA IgG assay is for the laboratory diagnosis of EBV-associated infectious mononucleosis in individuals with signs and symptoms consistent with infectious mononucleosis.

c) Special condition for use statement(s):
The device is for prescription use only

d) Special instrument Requirements:
NA

I. Device Description:
Indirect chemiluminescence immunoassay

J. Substantial Equivalence Information:

a) Predicate device name(s):
DiaSorin ETI-VCA-G Kit
DiaSorin ETI-EBV-M reverse Kit
DiaSorin ETI-EBNA-G Kit

b) Predicate K number(s):
K946159
K946157
K946158

{2}

Page 3 of 14

Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Same target population. | DiaSorin LIAISON® VCA IgG, LIAISON® VCA IgM, and the LIAISON® EBNA IgG assay (K041020) | DiaSorin ETI-VCA-G Kit
DiaSorin ETI EBV-M Reverse Kit, DiaSorin ETI-EBNA-G Kit |
| | Test persons who have symptoms of infectious mononucleosis | Test persons who have symptoms of infectious mononucleosis |
| Differences | | |
| Item | Device | Predicate |
| Different Methodology | Indirect chemiluminescence immunoassay | IgG Indirect ELISA |
| Different Indications for Use | Qualitative | Qualitative/Semiquantitative |
| Capture Reagent | Magnetic particles coated with antigen | Microtiter plate wells coated with antigen |
| Detector antibody Species | Mouse | Goat |
| Equivocal Zone | Yes | No |

K. Standard/Guidance Document Referenced (if applicable):

L. Test Principle:

The method for the qualitative determination of specific IgG or IgM to EBV viral antigens (VCA, EBNA) is an indirect chemiluminescence immunoassay (CLIA). The principal components of the test are magnetic particles (solid phase) coated with EBV synthetic peptides and a conjugate of mouse monoclonal antibody to human IgG or IgM linked to an isoluminol derivative (isoluminol-antibody conjugate). During the first incubation, VCA or EBNA antibodies, if present, bind to the solid phase. During the second incubation, the antibody conjugate reacts with EBV IgG or IgM antibodies that are bound to the solid phase. After each incubation step, unbound material is removed with wash cycles. Subsequently, the starter reagents are added and a flash chemiluminescence reaction is thus induced. The light signal, and hence the amount of

{3}

isoluminol-antibody conjugate, is measured by a photomultiplier as relative light units (RLU) and is indicative of the presence or absence of EBV VCA or EBNA IgG, or EBV VCA IgM antibodies.

# M. Performance Characteristics (if/when applicable):

# Analytical performance:

# a) Precision/Reproducibility:

An assay reproducibility study was conducted at two external US laboratories and at DiaSorin. A coded panel comprised of 9 frozen repository serum samples was prepared by DiaSorin and provided to each site for testing by the LIAISON® VCA IgG, VCA IgM and by the LIAISON® EBNA assay. The panel members were prepared to represent low to mid positive analyte levels. All panel members were divided into aliquots and stored frozen prior to testing. The same coded panel was tested at all three sites, in three replicates per run for ten runs. The results are summarized in the following tables.

LIAISON VCA IgG

| ID# | N | mean U/mL | within run S.D. | within run %CV | between run S.D. | between run %CV | between site S.D. | between site %CV | overall S.D. | overall %CV |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| VGS1 | 90 | 266.6 | 9.86 | 3.68 | 23.56 | 7.23 | 12.98 | 4.87 | 25.19 | 9.45 |
| VGS2 | 90 | 52.9 | 1.89 | 3.83 | 3.75 | 5.43 | 2.81 | 5.32 | 4.35 | 8.22 |
| VGS3 | 90 | 145.5 | 7.55 | 5.78 | 11.69 | 7.41 | 4.71 | 3.23 | 15.37 | 10.56 |
| VG1 | 90 | 31.6 | 0.71 | 2.31 | 2.94 | 6.36 | 2.64 | 8.35 | 2.99 | 9.45 |
| VG2 | 90 | 52.2 | 1.09 | 2.06 | 2.88 | 5.43 | 0.97 | 1.82 | 3.07 | 5.89 |
| VG3 | 90 | 61.0 | 1.17 | 1.92 | 3.75 | 5.64 | 2.05 | 3.35 | 3.86 | 6.32 |
| VG4 | 90 | 69.2 | 1.58 | 2.31 | 4.56 | 5.38 | 3.28 | 4.74 | 4.79 | 6.92 |
| VG5 | 90 | 58.0 | 1.21 | 2.11 | 3.07 | 4.08 | 2.47 | 4.26 | 3.27 | 5.63 |
| VG6 | 90 | 49.6 | 1.23 | 2.46 | 3.05 | 5.76 | 1.46 | 2.94 | 3.25 | 6.55 |

LIAISON VCA IgM

| ID# | N | mean U/mL | within run S.D. | within run %CV | between run S.D. | between run %CV | between site S.D. | between site %CV | overall S.D. | overall %CV |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| EMS1 | 90 | 67.2 | 2.22 | 3.30 | 3.36 | 4.26 | 2.21 | 3.28 | 3.92 | 5.83 |
| EMS2 | 89 | 89.7 | 4.00 | 4.55 | 6.90 | 5.16 | 6.26 | 6.98 | 7.79 | 8.70 |
| EMS3 | 90 | <10.0 | 69.6* | 7.15* | 83.6* | 6.50* | 69.5* | 7.35* | 115.7* | 12.24* |
| EM1 | 90 | 36.4 | 2.79 | 7.88 | 4.93 | 5.50 | 5.43 | 14.92 | 5.52 | 15.16 |

{4}

| EM2 | 90 | 37.2 | 2.23 | 6.09 | 5.43 | 6.17 | 5.91 | 15.90 | 5.77 | 15.52 |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| EM3 | 90 | 79.5 | 6.22 | 7.79 | 12.04 | 5.63 | 13.16 | 16.55 | 13.25 | 16.67 |
| EM4 | 89 | 65.9 | 4.34 | 6.60 | 8.95 | 5.35 | 9.95 | 15.10 | 9.77 | 14.83 |
| EM5 | 89 | 37.1 | 2.65 | 7.27 | 4.25 | 6.92 | 4.14 | 11.16 | 4.88 | 13.18 |
| EM6 | 90 | 64.6 | 4.26 | 6.65 | 6.07 | 6.58 | 5.36 | 8.30 | 7.18 | 11.11 |

*EMS3 dose was below the reading range of the assay. Precision calculations are based on signal (RLU) for this sample.

LIAISON® EBNA IgG

| ID# | N | mean U/mL | within run S.D. | within run %CV | between run S.D. | between run %CV | between site S.D. | between site %CV | overall S.D. | overall %CV |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| EBS1 | 90 | 28.5 | 0.73 | 2.57 | 1.04 | 2.70 | 0.83 | 2.90 | 1.24 | 4.35 |
| EBS2 | 90 | 62.2 | 4.81 | 6.94 | 8.84 | 11.78 | 3.70 | 5.95 | 15.02 | 24.15 |
| EBS3 | 90 | 290.4 | 12.18 | 4.27 | 26.81 | 6.02 | 24.49 | 8.43 | 29.04 | 10.00 |
| EB1 | 90 | 72.2 | 1.81 | 2.48 | 3.32 | 2.82 | 3.12 | 4.32 | 3.72 | 5.15 |
| EB2 | 90 | 71.0 | 2.12 | 3.01 | 5.45 | 3.86 | 5.74 | 8.09 | 5.73 | 8.08 |
| EB3 | 90 | 52.6 | 1.71 | 3.27 | 3.06 | 4.30 | 2.53 | 4.81 | 3.45 | 6.55 |
| EB4 | 90 | 48.6 | 1.51 | 3.17 | 3.30 | 4.72 | 2.93 | 6.04 | 3.57 | 7.34 |
| EB5 | 90 | 59.9 | 1.76 | 2.98 | 3.10 | 4.35 | 2.18 | 3.63 | 3.53 | 5.88 |
| EB6 | 90 | 51.6 | 1.69 | 3.33 | 2.61 | 4.61 | 1.46 | 2.83 | 3.06 | 5.94 |

a. Linearity/assay reportable range: NA
b. Traceability, Stability, Expected values (controls, calibrators, or method): NA
c. Detection limit: NA

# d. Analytical specificity:

The cross-reactivity studies for the LIAISON® EBV assays were designed to evaluate potential interference from IgG immunoglobulins directed against closely-related members of the herpes virus family (HSV-1, HSV-2, VZV, CMV, HHV6), from other organisms that may cause symptoms similar to EBV (Toxoplasma gondii,

{5}

rubella virus) and from other conditions that may result from atypical immune system activity (rheumatoid factor (RF)). Samples for these studies were selected using commercially available devices.

| Organism / condition | Number of Samples | Positive LIAISON® VCA IgG Result |
| --- | --- | --- |
| CMV IgG | 16 | (1/16) |
| VZV IgG | 7 | (0/7) |
| HSV-1 IgG | 18 | (2/18) |
| HSV-2 IgG | 3 | (0/3) |
| HHV6 IgG | 3 | (0/3) |
| Toxoplasma gondii IgG | 8 | (1/8) |
| Rubella virus IgG | 30 | (0/30) |
| RF | 4 | (0/4) |
| Total | 89 | (4/89) |

Four specimens out of 89 total specimens tested from the disease panel were positive. Due to the limited availability of certain samples, the possibility of cross-reactivity cannot be excluded. The user is advised to perform other EBV serology assays to confirm EBV-associated infectious mononucleosis.

| Organism / condition | Number of Samples | Positive LIAISON® EBV IgM Result |
| --- | --- | --- |
| CMV IgM | 29 | (2/29) |
| VZV IgM | 16 | (1/16) |
| HSV-1 IgM | 2 | (0/2) |
| HSV-2 IgM | 7 | (0/7) |
| Hepatitis A virus IgM | 10 | (0/10) |
| Hepatitis B virus (core) IgM | 23 | (0/23) |
| Toxoplasma gondii IgM | 12 | (0/12) |
| Rubella virus IgM | 11 | (0/11) |
| Rubeola virus IgM | 3 | (0/3) |
| Mumps virus IgM | 2 | (1/2) |
| RF | 6 | (0/6) |
| ANA Ig | 10 | (0/10) |
| Total | 131 | (4/131) |

{6}

Four specimens out of 131 total specimens tested from the disease panel were positive. Due to the limited availability of certain samples, the possibility of cross-reactivity cannot be excluded. Other EBV serology assays should be performed to confirm EBV-associated infectious mononucleosis.

| Organism / condition | Number of Samples | Positive LIAISON® EBNA IgG Result |
| --- | --- | --- |
| CMV IgG | 19 | (0/19) |
| VZV IgG | 7 | (0/7) |
| HSV-1 IgG | 19 | (1/19) |
| HSV-2 IgG | 3 | (0/3) |
| HHV6 IgG | 1 | (0/1) |
| Toxoplasma gondii IgG | 7 | (0/7) |
| Rubella virus IgG | 30 | (1/30) |
| ANA | 1 | (0/1) |
| RF | 4 | (0/4) |
| Total | 91 | (2/91) |

Two specimens out of 91 total specimens tested from the disease panel were positive. Due to the limited availability of certain samples, the possibility of cross-reactivity cannot be excluded. The user is advised to perform other EBV serology assays to confirm EBV-associated infectious mononucleosis.

# a. Assay cut-off:

The cutoff for the LIAISON® VCA IgG assay was determined during European clinical trials in which 1133 samples were tested at four separate sites. The LIAISON® EBV IgM assay was determined using 1332 samples and the LIAISON® EBNA IgG assay cutoff was determined using 1166 samples. The samples were drawn from several different populations, including seronegative subjects, subjects with primary EBV infection, past EBV infection, suspected chronic EBV infection and reactivated EBV infection and from an apparently healthy adult population.

# b. Comparison studies:

# a. Method comparison with predicate device:

DiaSorin LIAISON® EBV assays were compared to the DiaSorin ETI-EBV assays.

# Matrix comparison:

NA

{7}

Page 8 of 14

c. Clinical studies:

a. Clinical sensitivity: NA
b. Clinical specificity: NA
c. Other clinical supportive data (when a and b are not applicable):

# PERFORMANCE CHARACTERISTICS

A total of 893 samples was tested – 823 prospectively and 70 retrospectively collected. The prospective samples represented 823 samples from subjects sent to the laboratory for EBV testing. The retrospective samples represented 70 samples from patients positive for VCA IgM. The testing was performed at three sites – a hospital, a donor laboratory, and at DiaSorin. All samples were tested with the DiaSorin LIAISON® EBV Assays and the DiaSorin ETI-EBV ELISAs.

# Retrospective Samples: VCA IgM-positive Samples

Using the results for the retrospective samples in three reference assays (VCA IgG, EBNA-1 IgG and VCA IgM ELISA), the samples were grouped into serological categories. Indeterminate is defined as a serological category which does not fall into the typical EBV marker pattern for EBV seronegative, acute or past infection. The profiles and number of occurrences are presented in the following table:

| | VCA IgG | VCA IgM | EBNA-1 IgG | Total |
| --- | --- | --- | --- | --- |
| EBV seronegative | – | – | – | 0 |
| Acute infection | + | + | – | 9 |
| Past infection | + | – | + | 0 |
| Indeterminate | | | | |
| VCA IgG only | + | – | – | 0 |
| VCA IgM only | – | + | – | 0 |
| EBNA IgG only | – | – | + | 0 |
| Convalescent | + | + | + | 61 |

Based on these serological classifications, the LIAISON® VCA IgG results for the retrospective samples were compared with those obtained with the reference assay (VCA IgG ELISA).

{8}

Based on these serological classifications, the LIAISON® EBV IgM results for the retrospective samples were compared with those obtained with the reference assay (VCA IgM ELISA).

| | Percent Agreement | | 95% confidence interval |
| --- | --- | --- | --- |
| EBV | N/A | | N/A |
| Acute infection | 77.8% | (7/9) | 40.0 – 97.2% |
| Past infection | N/A | | N/A |
| Indeterminate | 98.4% | (60/61) | 91.2 – 100.0% |
| Overall | 95.7% | (67/70) | 88.0 – 99.1% |

Based on these serological classifications, the LIAISON® EBV IgG results for the retrospective samples were compared with those obtained with the reference assay (EBNA-1 IgG ELISA).

| | Percent Agreement | | 95% confidence interval |
| --- | --- | --- | --- |
| EBV | N/A | | N/A |
| Acute infection | 55.6% | (5/9) | 21.2 – 86.3% |
| Past infection | N/A | | N/A |
| Indeterminate | 100.0% | (61/61) | 95.2 – 100.0% |
| Overall | 94.3% | (66/70) | 86.0 – 98.4% |

# Prospective samples: Subjects Sent to the Laboratory for EBV Testing

Using the results for the prospective samples in three reference assays (VCA IgG, EBNA-1 IgG and VCA IgM ELISA), the samples were grouped into serological

{9}

categories. Indeterminate is defined as a serological category which does not fall into the typical EBV marker pattern for EBV seronegative, acute or past infection. Four samples tested in the LIAISON® VCA IgG assay had insufficient volume for the entire test profile and are omitted from this analysis. The profiles and number of occurrences are presented in the following table:

| | VCA IgG | VCA IgM | EBNA-1 IgG | Total |
| --- | --- | --- | --- | --- |
| EBV seronegative | - | - | - | 62 |
| Acute infection | + | + | - | 29 |
| Past infection | + | - | + | 573 |
| Indeterminate | | | | |
| VCA IgG only | + | - | - | 67 |
| VCA IgM only | - | + | - | 5 |
| EBNA IgG only | - | - | + | 10 |
| Convalescent | + | + | + | 73 |

Based on these serological classifications, the LIAISON® VCA IgG results for the prospective samples were compared with those obtained with the reference assay (VCA IgG ELISA).

| | Percent Agreement | | 95% confidence interval |
| --- | --- | --- | --- |
| EBV | 98.4% | (61/62) | 91.3 – 100.0% |
| Acute infection | 82.8% | (24/29) | 64.2 – 94.2% |
| Past infection | 98.3% | (563/573) | 96.8 – 99.2% |
| Indeterminate | 89.7% | (139/155) | 83.8 – 94.0% |
| Overall | 96.1% | (787/819) | 94.5 – 97.3% |

Based on these serological classifications, the LIAISON® EBV IgM results for the prospective samples were compared with those obtained with the reference assay (VCA IgM ELISA).

| | Percent Agreement | | 95% confidence interval |
| --- | --- | --- | --- |
| EBV | 93.5% | (58/62) | 84.3 – 98.2% |
| Acute infection | 100.0% | (29/29) | 90.2 – 100.0% |
| Past infection | 95.3% | (546/573) | 93.2 – 96.9% |

{10}

Based on these serological classifications, the LIAISON® EBNA IgG results for the prospective samples were compared with those obtained with the reference assay (EBNA-1 IgG ELISA).

| | Percent Agreement | | 95% confidence interval |
| --- | --- | --- | --- |
| EBV | 100.0% | (62/62) | 95.3 – 100.0% |
| Acute infection | 100.0% | (29/29) | 90.2 – 100.0% |
| Past infection | 98.1% | (562/573) | 96.6 – 99.0% |
| Indeterminate | 81.9% | (127/155) | 75.0 – 87.7% |
| Overall | 95.2% | (780/819) | 93.6 – 96.6% |

d. Clinical cut-off:

NA

e. Expected values/Reference range:

The LIAISON® VCA IgG assay was tested with prospectively collected samples from subjects sent to the laboratory for EBV testing $(n = 823)$ to evaluate the prevalence of IgG antibodies to VCA in these populations. The subjects sent to the laboratory for EBV testing were $61.7\%$ female (508), $28.1\%$ male (231) and $10.2\%$ unknown (84) and represented the mid-Atlantic and Northeastern US.

The distribution of results for IgG antibodies to VCA in this population as determined by the LIAISON® VCA IgG Assay is summarized in the following table.

| | N | Negative | Equivocal | Positive | Prevalence |
| --- | --- | --- | --- | --- | --- |
| Total | 823 | 97 | 5 | 721 | 87.6% |
| Gender | | | | | |
| Female | 508 | 56 | 2 | 450 | 88.6% |
| Male | 231 | 32 | 3 | 196 | 84.8% |
| Unknown | 84 | 9 | 0 | 75 | 89.3% |
| Age (years) | | | | | |
| ≤ 18 | 173 | 36 | 2 | 135 | 78.0% |
| 18-24 | 100 | 18 | 0 | 95 | 89.3% |
| 25-34 | 100 | 18 | 0 | 95 | 89.3% |
| 35-44 | 100 | 18 | 0 | 95 | 89.3% |
| 45-54 | 100 | 18 | 0 | 95 | 89.3% |
| ≥ 55 | 100 | 18 | 0 | 95 | 89.3% |

{11}

The LIAISON® EBV IgM Assay was tested with prospectively collected samples from subjects sent to the laboratory for EBV testing $(n = 819)$ to evaluate the prevalence of IgM antibodies to VCA in these populations. The subjects sent to the laboratory for EBV testing were $61.8\%$ female (506), $28.0\%$ male (229) and $10.3\%$ unknown (84) and represented the mid-Atlantic and Northeastern US.

The distribution of results for IgM antibodies to VCA in this population as determined by the LIAISON® EBV IgM Assay is summarized in the following table.

| | N | Negative | Equivocal | Positive | Prevalence |
| --- | --- | --- | --- | --- | --- |
| Total | 819 | 719 | 14 | 86 | 10.5% |
| Gender | | | | | |
| Female | 506 | 453 | 8 | 45 | 8.9% |
| Male | 229 | 194 | 3 | 32 | 14.0% |
| Unknown | 84 | 72 | 3 | 9 | 10.7% |
| Age (years) | | | | | |
| ≤ 18 | 172 | 138 | 2 | 32 | 18.6% |
| < 10 | 29 | 23 | 0 | 6 | 20.7% |
| 10 – 19 | 182 | 148 | 2 | 32 | 17.6% |
| 20 – 29 | 190 | 159 | 6 | 25 | 13.2% |
| 30 – 39 | 95 | 90 | 2 | 3 | 3.2% |
| 40 – 49 | 77 | 71 | 2 | 4 | 5.2% |
| 50 – 59 | 59 | 56 | 1 | 2 | 3.4% |
| 60 – 69 | 32 | 32 | 0 | 0 | 0.0% |
| ≥ 70 | 25 | 24 | 0 | 1 | 4.0% |
| Unknown | 130 | 116 | 1 | 13 | 10.0% |

{12}

The LIAISON® EBNA IgG Assay was tested with prospectively collected samples from subjects sent to the laboratory for EBV testing $(n = 823)$ to evaluate the prevalence of IgG antibodies to EBNA in these populations. The subjects sent to the laboratory for EBV testing were $61.7\%$ female (508), $28.1\%$ male (231) and $10.2\%$ unknown (84) and represented the mid-Atlantic and Northeastern US.

The distribution of results for IgG antibodies to EBNA in this population as determined by the LIAISON® EBNA IgG Assay is summarized in the following table.

| | N | Negative | Equivocal | Positive | Prevalence |
| --- | --- | --- | --- | --- | --- |
| Total | 823 | 177 | 5 | 641 | 77.9% |
| Gender | | | | | |
| Female | 508 | 96 | 1 | 411 | 80.9% |
| Male | 231 | 64 | 3 | 164 | 70.0% |
| Unknown | 84 | 17 | 1 | 66 | 78.6% |
| Age (years) | | | | | |
| ≤ 18 | 173 | 63 | 2 | 108 | 62.4% |
| < 10 | 29 | 11 | 1 | 17 | 58.6% |
| 10 – 19 | 183 | 65 | 1 | 117 | 63.9% |
| 20 – 29 | 190 | 41 | 0 | 149 | 78.4% |
| 30 – 39 | 95 | 8 | 1 | 86 | 90.5% |
| 40 – 49 | 78 | 5 | 1 | 72 | 92.3% |
| 50 – 59 | 59 | 7 | 1 | 51 | 86.4% |
| 60 – 69 | 32 | 5 | 0 | 27 | 84.4% |
| ≥ 70 | 27 | 6 | 0 | 21 | 77.8% |
| Unknown | 130 | 29 | 0 | 101 | 77.7% |

# N. Proposed labeling:

The labeling is sufficient and it satisfies the requirement of 21 CFR Part 809.10.

WARNINGS: Assay interference due to circulating antibodies against HIV and Hepatitis A, Hepatitis B and Hepatitis C viruses has not been evaluated. The user is responsible for establishing cross-reactivity performance with these infectious agents

The recommended LIAISON® EBNA IgG quality control material contains a $5\%$ serum matrix. It may not adequately control the DiaSorin LIAISON® EBNA IgG assay for serum specimens. The user must provide quality control material for

{13}

Page 14 of 14

serum specimens. Alternative materials for the control of serum specimens include commercial quality control materials or your laboratory's own pooled serum specimens. Choose control levels that check assay performance at all clinically relevant points (e.g., assay cutoff). The recommendation is to run a positive and negative control close (± 50%) to the assay's decision point. It is the responsibility of the user to validate the use of alternative control materials with this assay and to establish appropriate control ranges

O. Conclusion:
The submitted material in this premarket notification is complete and supports a substantial equivalence decision.