**Summary of Safety and Effectiveness Data**

I.  **General Information**

Device Generic Name: A solid-phase chemiluminescent enzyme immunoassay
for the qualitative detection of hepatitis B surface antigen (HBsAg)

Device Trade Name: IMMULITE HBsAg

> IMMULITE 2000 HBsAg
>
> IMMULITE HBsAg Confirmatory Kit

Applicant's Name and Address: Diagnostic Products Corporation

> 5700 West 96^th^ Street

Los Angeles, California 90045-5597

Premarket Approval Application (PMA) Number: P010050

Date of Panel Recommendation: None

Date of Notice of Approval to the Applicant: July 26, 2002

II. **INDICATIONS FOR USE**

IMMULITE HBsAg

IMMULITE HBsAg is a solid-phase chemiluminescent enzyme immunoassay
designed for use on the IMMULITE automated immunoassay analyzer for the
qualitative detection of hepatitis B surface antigen (HBsAg) in human
serum or plasma (EDTA, heparin, citrate). It is intended for in vitro
diagnostic use for the laboratory diagnosis of acute and chronic
hepatitis B virus infections in conjunction with other serological and
clinical information. In addition, this assay may be used to screen for
hepatitis B infection in pregnant women to identify neonates who are at
high risk of acquiring HBV during the perinatal period.

######## IMMULITE 2000 HBsAg

IMMULITE 2000 HBsAg is a solid-phase chemiluminescent enzyme immunoassay
designed for use on the IMMULITE 2000 automated immunoassay analyzer for
the qualitative detection of hepatitis B surface antigen (HBsAg) in
human serum or plasma (EDTA, heparin, citrate). It is intended for in
vitro diagnostic use for the laboratory diagnosis of acute and chronic
hepatitis B virus infections in conjunction with other serological and
clinical information. In addition, this assay may be used to screen for
hepatitis B infection in pregnant women to identify neonates who are at
high risk of acquiring HBV during the perinatal period.

**IMMULITE HBsAg Confirmatory**

IMMULITE HBsAg Confirmatory is intended for in vitro diagnostic use in
conjunction with the IMMULITE HBsAg or the IMMULITE 2000 HBsAg assays
for the confirmation of the presence of hepatitis B surface antigen
(HBsAg) in human serum or plasma (EDTA, heparin, citrate) that was
repeatedly positive when tested by IMMULITE or IMMULITE 2000 HBsAg.

III. **DESCRIPTION OF DEVICE**

IMMULITE and IMMULITE 2000 HBsAg kits and the IMMULITE HBsAg
Confirmatory Kit are the subjects of this PMA. The data and information
are presented separately for all three.

The IMMULITE and IMMULITE 2000 HBsAg kits are solid phase, two step
chemiluminescent enzyme immunoassays designed for use on the automated
IMMULITE and IMMULITE 2000 analyzers, for the qualitative detection of
hepatitis B surface antigen (HBsAg) in human serum or plasma. The kits
are intended for in vitro use as an aid in the laboratory diagnosis of
acute and chronic hepatitis B virus infections. The kits' solid phase is
a polystyrene bead coated with an antibody directed against the
hepatitis B surface antigen (anti-HBs). The patient sample and a
protein-based buffer are simultaneously introduced into the Test Unit
and incubated for approximately 30 minutes at 37 ^o^C with intermittent
agitation. During this time, HBsAg in the patient sample binds to the
anti-HBs coated bead. Unbound serum is then removed by a centrifugal
wash. An alkaline phosphatase-labeled anti-HBs is introduced, and the
reaction tube is incubated with agitation for another 30 minute cycle.
The unbound enzyme conjugate is removed by a centrifugal wash. After the
wash, a chemiluminescent substrate is added, and the reaction tube is
incubated with agitation for a further 5 -- 10 minutes.

The chemiluminescent substrate is a phosphate ester of adamantyl
dioxetane which undergoes hydrolysis in the presence of alkaline
phosphatase to yield an unstable intermediate. The continuous production
of this intermediate results in the sustained emission of light. The
bound complex and the resulting photon output, measured as cps by the
photomultiplier tube, is related to the presence of HBsAg in the sample.
A qualitative result is then obtained by comparing the patient results
to an established cutoff.

In assays for HBsAg where the instrument response signal is directly
related to the antigen concentration, the presence of HBsAg in a sample
can be confirmed by demonstrating a significant reduction in signal
following specific antibody neutralization. The signal reduction
reflects a reduction in binding of HBsAg to the solid phase antibodies
in the presence of blocking antibodies. A sufficiently decreased signal
relative to that of a control sample confirms a positive HBsAg result.

The IMMULITE HBsAg Confirmatory kit is used in conjunction with the
IMMULITE and IMMULITE 2000 HBsAg assays to confirm the presence of HBsAg
in a patient sample that has initially tested reactive for the antigen.
In the confirmatory procedure, an undiluted sample and a 1:500 dilution
of the sample are each divided into two aliquots. One aliquot is
combined with a blocking reagent containing goat anti-HBs, the other
with a control reagent which lacks the antibody.

For suitably diluted HBsAg positive samples, in the aliquots containing
the blocking reagent, most of the HBsAg present in solution binds to the
blocking antibody and does not bind to the coated bead; whereas, in the
aliquots containing the control reagent, any HBsAg present in the sample
remains free to bind to the coated bead. The original patient sample is
confirmed positive for the presence of HBsAg if the signal from either
the undiluted or diluted aliquots of the blocked aliquots is at least
50% less than the signal from the corresponding unblocked (control)
aliquot.

IV. **[Contraindications, WARNINGS AND PRECAUTIONS]{.mark}**

[]{.mark}

[There are no known contraindications for the]{.mark} IMMULITE HBsAg,
IMMULITE 2000 HBsAg and the IMMULITE HBsAg Confirmatory Kit

> []{.mark}

Warning and precautions for users of the IMMULITE HBsAg, IMMULITE2000
HBsAg and the IMMULITE HBsAg Confirmatory assays are stated in the
product labeling.

**V. ALTERNATIVE PRACTICES OR PROCEDURES**

There are currently a variety of commercially available, FDA licensed or
approved serological tests for the detection of HBsAg. Assays have been
developed using radioimmunoassay or enzyme immunoassay methodologies.
When the test results are used in combination with other serological
markers expressed during the three phases of incubation, the presence of
hepatitis B virus may be determined.

**VI. MARKETING HISTORY**

IMMULITE HBsAg and IMMULITE 2000 HBsAg have been marketed
internationally as an aid in the determination of hepatitis B virus
infection status since July 1996. IMMULITE and IMMULITE 2000 HBsAg have
received European Union CE Mark approval and have been marketed in
Europe since June 2001.

**VII. POTENTIAL ADVERSE EFFECTS OF DEVICE ON HEALTH**

There are no direct adverse effects on the health of the patient if the
devices are used according to the instructions in the package insert.
However, failure of the device to perform as indicated or human error in
use of the device may lead to inaccurate results and a misdiagnosis. A
false positive result using an HBsAg assay may be considered a patient
or public health concern if the positive result is not done in a
clinical lab setting where a positive test is followed up with
supplemental testing. Either additional HBV marker testing is performed
or an HBsAg positive result is confirmed by neutralization. An exception
to this is using HBsAg tests to screen pregnant women for the presence
of HBsAg. This testing helps to determine if a neonate is at high risk
of acquiring HBV during the prenatal period. Pregnant women are tested
during an early prenatal visit. If they are HBsAg nonreactive during
this testing, and at high risk for HBV infection, they are re-tested
during the third trimester. If the result is positive, it is recommended
that hepatitis B immune globulin (HBIG) and vaccine be provided to the
newborn within 12 hours of birth. If an assay is false positive and the
newborn receives HBIG, the new born would be exposed to the risks of
receiving a human source product.

The risks of a false negative result in a diagnostic setting are highest
when testing pregnant women because HBsAg may be the only marker used.
If the result is negative then the child is vaccinated within 2 months
of birth. If the result is incorrect (false negative), the neonate is at
a higher risk of acute and chronic HBV infection, since HBIG and vaccine
would not be provided within 12 hours of birth.

In addition, from time to time false negative results due to gene
mutation have been reported for HBsAg assays produced by a number of
different manufacturers. Because of the complexity of the mutations that
can occur, no manufacturer can guarantee to detect all infectious donors
and patients.

A false positive result using an HBsAg confirmatory neutralization
procedure is not considered a patient or public health concern because
in order for a false positive to occur, the control sample
(non-neutralized result) and the percent neutralization (in neutralized
tube) would both have to be incorrect for a reported false positive
result. If this situation were to occur, the implications would be the
same as described for false positive results for HBsAg assays.

A false negative result using an HBsAg confirmatory neutralization
procedure could occur if the neutralized sample were incorrect either
due to a falsely increased signal with the neutralized sample, or due to
some other malfunction, laboratory or technician error when assayed. A
falsely increased signal could be interpreted as a failure to
neutralize. If this situation were to occur, the implications would be
the same as described for false negative results for HBsAg.

VIII. **SUMMARY OF NON CLINICAL STUDIES**

> **Analytical Specificity**

Analytical specificity was evaluated at two clinical sites in the United
States and at one European site. In the United States, serum specimens
from 17 subgroups of patients with potentially cross-reacting
microorganisms or conditions were tested by IMMULITE HBsAg and a
commercially available enzyme immunoassay for HBsAg (Kit A). These
results are listed in the package insert.

In the European study, eight specimens from antinuclear antibody (ANA)
positive patients and 30 from patients with positive rheumatoid factor
(RF) were tested by IMMULITE HBsAg. IMMULITE HBsAg test results were all
negative for these

38 specimens.

The specimens were also tested by IMMULITE 2000 HBsAg, at the same
sites. The test results are presented in the package inserts.

In the European study, seven specimens from antinuclear antibody (ANA)
positive

patients and 24 from patients with positive rheumatoid factor (RF) were
tested by IMMULITE 2000 HBsAg. IMMULITE 2000 HBsAg test results were all
negative for these 31 specimens.

**Analytical Sensitivity**

Based on studies with serial dilution of WHO International Standard for
Hepatitis B Surface Antigen (NIBSC 80/549), the threshold sensitivity
(last positive dilution) for IMMULITE and IMMULITE 2000 HBsAg is 0.063
IU/mL. The 95% confidence interval at this level (0.063 IU/mL) is
0.040 - 0.086 IU/mL.

**Effects of Bilirubin, Lipemia and Hemolysis**

To simulate moderate and severe icterus, different volumes of each of 6
patient samples ranging from very negative to high positive hepatitis B
surface antigen values were pipetted into lyophilized unconjugated
bilirubin to achieve 3 levels of bilirubin concentrations (10 and 20
mg/dL) for each sample. The spiked and unspiked samples were assayed by
the IMMULITE and IMMULITE 2000 HBsAg assays. In the IMMULITE HBsAg
tests, the spiked samples had averages of 96%, 99% and 99% recovery for
all samples for 10 and 20 mg/dL of bilirubin concentrations,
respectively. In the IMMULITE 2000 HBsAg tests, the spiked samples had
averages of 101%, 98% and 101% recovery for all samples for 10 and 20
mg/dL bilirubin concentrations, respectively. This study demonstrated
that the measurement of hepatitis B surface antigen was not affected by
the presence of bilirubin up to 20 mg/dL.

To simulate mild, moderate and severe hemolysis, the same 6 samples were
spiked with hemolysate to achieve final hemoglobin levels of 168, 252
and 504 mg/dL. The samples were assayed, both spiked and unspiked, by
the IMMULITE and IMMULITE 2000 HBsAg assays. Expected kCPS for each
sample was calculated based on the spiking factor and compared with the
observed kCPS. In the IMMULITE HBsAg tests, the spiked samples had
averages of 115%, 126% and 144% recovery for all samples for 168, 252
and 504 mg/dL hemoglobin concentrations, respectively. In the IMMULITE
2000 HBsAg tests, the spiked samples had averages of 111%, 114% and 136%
recovery for all samples for 168, 252 and 504 mg/dL hemoglobin
concentrations, respectively. Since increases of responses (kcps) were
observed in the spiked samples (especially the low samples) with the
increase of hemoglobin, it was concluded that the measurement of
hepatitis B surface antigen may be affected by the presence of red blood
cells. The results of hemolyzed samples should be interpreted with
caution.

The same 6 samples were each spiked with 4 levels of triglycerides at
500, 1000, 2000 and 3000 mg/dL to evaluate the lipemia effect on the
IMMULITE and IMMULITE 2000 HBsAg assays. Unspiked and spiked samples
were tested by the IMMULITE and IMMULITE 2000 HBsAg assays. Since
increases of responses (kcps) in the spiked samples correlated with the
increase of triglyceride levels, it was concluded that the measurement
of hepatitis surface antigen may be affected by the presence of
triglycerides. The use of an ultracentrifuge is recommended to clear
lipemic samples.

**Hook Effect and Carryover**

The hook effect study demonstrated that the IMMULITE and IMMULITE 2000
HBsAg assays did not have a hook effect up to at least 2000 times the
titer of the cutoff.

The carryover study demonstrated that the IMMULITE and IMMULITE 2000
HBsAg assays did not exhibit a carryover phenomenon when samples were
preceded by a sample with very high antigen titers.

**Interfering Substances**

A study was conducted to evaluate the effects of interfering substances
on IMMULITE and IMMULITE 2000 HBsAg assays. Potential interfering
substances that included common serum constituents, chemotherapeutic and
other drugs were spiked into serum samples with 5 or 6 different levels
of HBsAg. Listed below are the substances and their test levels
(concentration).

![](media/image1.wmf){width="2.6631944444444446in"
height="2.6131944444444444in"}

This study demonstrated that the detection of hepatitis B surface
antigen by IMMULITE and IMMULITE 2000 HBsAg was not affected by the
presence of any of the interfering substances listed up to the levels
tested.

**Effects of Anticoagulants**

The measurement of HBsAg is not affected by the presence of heparin,
sodium citrate, and EDTA anti-coagulants, as shown in a study that
included 60 specimens collected into plain, heparinized, sodium citrate,
and EDTA vacutainer tubes. By regression:

IMMULITE HBsAg (ratio):

(EDTA) = 1.01 (Serum) + 0.41 r = 1.00

(Na Citrate) = 1.03 (Serum) + 0.74 r = 1.00

(Heparin) = 1.03 (Serum) + 1.10 r = 1.00

IMMULITE 2000 HBsAg (ratio):

(EDTA) = 0.98 (Serum) + 1.52 r = 1.00

(Na Citrate) = 0.96 (Serum) + 0.83 r = 1.00

(Heparin) = 0.99 (Serum) + 1.93 r = 1.00

In another study conducted in the northwestern United States, blood
samples from 18 chronic hepatitis B patients at different stages of the
disease were drawn into plain tubes and blood collection tubes that
contained Na citrate, EDTA or heparin. All samples were assayed by
IMMULITE and IMMULITE 2000 HBsAg. Test results for serum samples and
plasma samples were all consistent for all 18 patients, yielding 100%
agreement between serum and the three anticoagulant samples for both
assays.

**Precision**

Serum samples were assayed in duplicate over the course of 20 days, two
runs per day, for a total of 40 runs and 80 replicates at three US
sites.

### IMMULITE HBsAg Intrassay and Total Precision (ratio)

> Site 1

  ---- ------------- ------------- ---------------- ---------- ---------------
                     Intra-assay                    Total      

       Mean          SD            CV               SD         CV

  1    0.44          0.033         7.5%             0.039      8.9%

  2    0.44          0.056         12.6%            0.073      16.5%

  3    1.19          0.072         6.0%             0.078      6.5%

  4    1.23          0.087         7.1%             0.165      13.4%

  5    1.44          0.102         7.1%             0.126      8.7%

  6    2.96          0.173         5.9%             0.191      6.4%
  ---- ------------- ------------- ---------------- ---------- ---------------

### 

### Site 2

  ----- -------------- ------------- --------------- --------- ----------------
                       Intra-assay                   Total     

        Mean           SD            CV              SD        CV

  1     0.41           0.043         10.5%           0.051     12.2%

  2     0.42           0.059         14.1%           0.079     18.9%

  3     1.18           0.067         5.7%            0.118     10.0%

  4     1.20           0.059         4.9%            0.156     13.0%

  5     1.37           0.059         4.3%            0.180     13.1%

  6     2.86           0.163         5.7%            0.356     12.5%
  ----- -------------- ------------- --------------- --------- ----------------

### 

### Site 3

  --- --------------- ------------- ----------------- --------- --------------
                      Intra-assay                     Total     

      Mean            SD            CV                SD        CV

  1   0.44            0.051         11.8%             0.071     16.3%

  2   0.42            0.044         10.6%             0.056     13.4%

  3   1.14            0.089         7.8%              0.122     10.7%

  4   1.13            0.086         7.6%              0.166     14.7%

  5   1.43            0.062         4.3%              0.146     10.2%

  6   2.86            0.128         4.5%              0.322     11.3%
  --- --------------- ------------- ----------------- --------- --------------

### IMMULITE 2000 HBsAg Intrassay and Total Precision (ratio)

Site 1

  ----- ------------ ------------- -------------- ---------- ----------------
                     Intra-assay                  Total      

        Mean         SD            CV             SD         CV

  1     0.55         0.060         11.1%          0.074      13.6%

  2     0.53         0.042         7.8%           0.061      11.4%

  3     1.16         0.091         7.8%           0.098      8.4%

  4     1.20         0.074         6.2%           0.177      14.8%

  5     1.47         0.102         7.0%           0.214      14.6%

  6     2.56         0.149         5.8%           0.243      9.5%
  ----- ------------ ------------- -------------- ---------- ----------------

### 

### Site 2

  ----- ----------- ------------- --------------- ---------- ----------------
                    Intra-assay                   Total      

        Mean        SD            CV              SD         CV

  1     0.46        0.041         8.9%            0.052      11.2%

  2     0.48        0.059         12.0%           0.066      13.7%

  3     1.18        0.063         5.4%            0.093      7.9%

  4     1.18        0.059         5.0%            0.152      12.9%

  5     1.45        0.092         6.4%            0.161      11.1%

  6     2.80        0.149         5.3%            0.244      8.7%
  ----- ----------- ------------- --------------- ---------- ----------------

### Site 3

  ----- -------------- ------------- --------------- ---------- --------------
                       Intra-assay                   Total      

        Mean           SD            CV              SD         CV

  1     0.50           0.038         7.6%            0.059      11.7%

  2     0.50           0.051         10.2%           0.071      14.2%

  3     1.17           0.126         10.8%           0.171      14.7%

  4     1.18           0.053         4.5%            0.144      12.2%

  5     1.48           0.091         6.1%            0.129      8.7%

  6     2.70           0.117         4.3%            0.406      15.0%
  ----- -------------- ------------- --------------- ---------- --------------

IMMULITE HBsAg Lot-to-Lot and Site-to-Site:

  ----- ------------ ------------ ---------------- -------------- ----------------
                     Lot-to-Lot                    Site-to-Site   

        Mean         SD           CV               SD             CV

  1     0.47         0.062        13.2%            0.063          13.4%

  2     0.47         0.069        14.7%            0.069          14.9%

  3     1.17         0.095        8.1%             0.099          8.5%

  4     1.19         0.157        13.2%            0.165          13.8%

  5     1.50         0.181        12.1%            0.182          12.1%

  6     2.85         0.266        9.3%             0.273          9.6%
  ----- ------------ ------------ ---------------- -------------- ----------------

IMMULITE 2000 HBsAg Site-to-Site Precision:

#### 

+-------------+---------------+---------------------+-----------------+
|             |               | Site-to-Site        |                 |
+-------------+---------------+---------------------+-----------------+
|             | Mean          | SD                  | CV              |
+-------------+---------------+---------------------+-----------------+
| 1           | 0.50          | 0.070               | 13.9%           |
+-------------+---------------+---------------------+-----------------+
| 2           | 0.51          | 0.069               | 13.7%           |
+-------------+---------------+---------------------+-----------------+
| 3           | 1.17          | 0.126               | 10.7%           |
+-------------+---------------+---------------------+-----------------+
| 4           | 1.18          | 0.158               | 13.3%           |
|             |               |                     |                 |
| 5           | 1.47          | 0.172               | 11.7%           |
+-------------+---------------+---------------------+-----------------+
| 6           | 2.69          | 0.321               | 11.9%           |
+-------------+---------------+---------------------+-----------------+

The IMMULITE 2000 HBsAg lot-to-lot precision was not evaluated on three
lots***.*** Because the reproducibility of the assay for lot-to-lot was
previously established, and Stability studies were performed using three
lots, and the data was acceptable, no additional reproducibility studies
were required.

EDTA, heparin, and sodium citrate samples were assayed in duplicate in
three runs on three days at three U.S. sites for three lots of IMMULITE
HBsAg and one lot of IMMULITE 2000 HBsAg. The median total variance of
coefficients (EDTA, 5.2%; heparin, 7.2%; sodium citrate, 5.9%)
demonstrated that these alternative sample types do not affect the
precision of IMMULITE HBsAg.

**Stability**

Stability studies for IMMULITE and IMMULITE 2000 HBsAg and HBsAg
Confirmatory were conducted by using 3 lots of IMMULITE HBsAg and 3 lots
of HBsAg Confirmatory, and one lot of IMMULITE 2000 HBsAg. The kits and
components were subjected to different storage/stress conditions to
simulate adverse conditions that might be encountered during shipment
and use at clinical laboratories, to establish the long-term
(shelf-life) claims, to approximate and support the real time stability
and to test the robustness of individual components.

These studies demonstrated that the performance of IMMULITE HBsAg,
IMMULITE 2000 HBsAg and IMMULITE HBsAg Confirmatory assays was not
affected if properly stored at package insert conditions for at least
720 days.

These studies also demonstrated that the performance of IMMULITE HBsAg,
IMMULITE 2000 HBsAg and IMMULITE HBsAg Confirmatory assays was not
affected following initial stresses (37°C, or --20°C) for at least 720
days.

**IX. Summary of Clinical Studies**

**Expected Values**

Individuals acutely infected with the hepatitis B virus will exhibit a
rapid rise (normally peaks at 12 weeks) and decline of detectable levels
of HBsAg between 1 and 4 months after exposure; usually through the
course of the clinical illness. HBsAg in acute infection can be detected
as early as 1 or 2 weeks and as late as 11 or 12 weeks. Of acutely
infected adults, 5--10% will become chronic carriers of the virus. In
chronic carriers, no significant decrease in HBsAg level is observed.
Because of variation in individual acute patients and the timing,
frequency and length of follow-up for each individual patient, the
percentage of specimens reported with positive results can be very
variable, as shown in the studies below.

Demographics and expected prevalence rates for different categories of
subjects (apparently healthy individuals, HBV Chronic, HBV Acute
patients) each of whom provided one specimen, from four clinical
studies, one in the northwestern United States (Study 1), two in the
southern United States (Study 3, using specimens from China, and Study
4) and one in Europe, are summarized in the package insert.

Five clinical studies were conducted to assess the performance of the
IMMULITE HBsAg (total number of subjects = 3268) and IMMULITE 2000 HbsAg
(total number of subjects = 2968) assays.

The participating subjects each contributed one specimen to the
analyses. The specimens were tested by FDA-approved or licensed
hepatitis B assays (Reference markers), IMMULITE HbsAg, and IMMULITE
2000 HbsAg. All specimens in the analyses were initial test results.

The data were analyzed following the assignment of specimen
classification based upon the reactive(+)/ nonreactive(-) patterns for
the HBV reference serological markers. Specimen classification was based
only on the HBV serological marker results for that particular specimen.
No other laboratory or clinical information was used in the disease
classification process.

**Study 1:** Conducted in the northwestern United States, this study
included 281 patients, consisting of 138 males and 88 females. Gender
for 55 subjects was not reported. These subjects had an average age of
43 years, ranging from 21 to 85 years. Distributions of the ethnicity of
the subjects are shown in the following table.

+-------------------------------+------------------+------------------+
| > Ethnicity                   | > *N*            | > \%             |
+-------------------------------+------------------+------------------+
| > African American            | > 15             | > 5.3%           |
+-------------------------------+------------------+------------------+
| > Caucasian                   | > 169            | > 60.1%          |
+-------------------------------+------------------+------------------+
| > Hispanic                    | > 2              | > 0.7%           |
+-------------------------------+------------------+------------------+
| > Asian                       | > 32             | > 11.4%          |
+-------------------------------+------------------+------------------+
| > Other                       | > 3              | > 1.1%           |
+-------------------------------+------------------+------------------+
| > Unknown                     | > 60             | > 21.4%          |
+-------------------------------+------------------+------------------+
| > Total                       | > 281            | > 100.0%         |
+-------------------------------+------------------+------------------+

These subjects included acute and chronic hepatitis B patients,
vaccinated individuals, pregnant women, and apparently healthy
individuals, and patients with potentially crossreactive substances and
medical conditions.

A total of 281 specimens were prospectively (n=92) and retrospectively
(n=189) collected and tested by FDA-approved or licensed hepatitis B
assays for six HBV serological markers (HbsAg, HbeAg, Anti-HBc IgM,
Anti-HBc, Anti-Hbe, and Anti-HBs). Characterization based on single
point specimens by the reference serological markers demonstrated 17
unique patterns:

  ----------------------------------------------------------------------------------------------
                     No. of     HBV                                                   
                     patients   Reference                                             
                                Markers                                               
  ------------------ ---------- ----------- -------- ---------- ---------- ---------- ----------
  Characterization              HbsAg       HbeAg    Anti-HBc   Anti-HBc   Anti-Hbe   Anti-HBs
  based on single                                    IgM                              
  point specimens                                                                     

  Acute              2          \+          --       --         --         --         --

  Acute              1          \+          \+       --         --         --         --

  Acute              32         \+          --       +/-        \+         \+         --

  Acute              34         \+          \+       +/-        \+         --         --

  Chronic            2          \+          --       --         \+         --         --

  Chronic            3          \+          +/-      --         \+         \+         \+

  Chronic            1          \+          --       --         \+         --         \+

  Chronic            1          \+          \+       +/-        \+         --         \+

  Early Recovery     16         --          --       +/-        \+         \+         \+

  Early Recovery     4          --          --       --         \+         \+         --

  Early Recovery     19         --          --       --         \+         +/-        --

  Early Recovery     1          --          --       \+         \+         +/-        \+

  HBV vaccine        27         --          --       --         --         --         \+
  response                                                                            

  Not previously     120        --          --       --         --         --         --
  infected                                                                            

  Recovered          16         --          --       --         +/-        --         \+

  Recovered          1          --          +/-      --         \+         --         \+

  Uninterpretable    1          --          \+       --         --         --         --
  ----------------------------------------------------------------------------------------------

Based on the above classifications the IMMULITE and IMMULITE 2000 HbsAg
results were compared to Kit A, a reference assay for the determination
of HbsAg.

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<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="9">Kit A</td>
<td colspan="2" rowspan="4">Total</td>
<td></td>
</tr>
<tr class="even">
<td colspan="3">+</td>
<td colspan="6">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="9">IML HbsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="3">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">67</td>
<td colspan="3">2</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">69</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">7</td>
<td colspan="3">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">7</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="2">0</td>
<td colspan="2">40</td>
<td colspan="2">40</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HBV Vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="2">0</td>
<td colspan="2">27</td>
<td colspan="2">27</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="2">1</td>
<td colspan="2">119</td>
<td colspan="2">120</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Recovered</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="2">1</td>
<td colspan="2">16</td>
<td colspan="2">17</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">74</td>
<td colspan="3">2</td>
<td colspan="2">2</td>
<td colspan="2">203</td>
<td colspan="2">281</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. The Total Positive agreement
is 97.4% (74/76) 95% with 95% CI of 90.8 to 99.7%. The Negative
agreement is 99.0% (203/205) with a 95% CI of 96.5 to 99.9%. The Total
agreement is 98.6% (277/281) with a 95% CI of 96.4 to 99.6%. The
complete data by category is presented in the package insert.

<table style="width:100%;">
<colgroup>
<col style="width: 32%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 17%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4">Total</td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML 2000 HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">67</td>
<td colspan="2">2</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">69</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">7</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">7</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">2</td>
<td colspan="2">38</td>
<td colspan="2">40</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HBV Vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">27</td>
<td colspan="2">27</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">6</td>
<td colspan="2">114</td>
<td colspan="2">120</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Recovered</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">16</td>
<td colspan="2">17</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">74</td>
<td colspan="2">2</td>
<td colspan="2">9</td>
<td colspan="2">196</td>
<td colspan="2">281</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. The Total Positive agreement
is 97.4% (74/76) with a 95% CI of 90.8 to 99.7%. The Negative agreement
is 95.6% (196/205) with a 95% CI of 91.8 to 98.0%. The Total agreement
is 96.1% (270/281) with a 95% CI of 93.1 to 98.0%. The complete data by
category is presented in the package insert.

**Study 2:** Conducted in the northeastern United States, this study
included 209 patients, consisting of 104 males and 103 females. Gender
for two patients was not reported. These patients had an average age of
47 years, ranging from newborn to 93 years. Distributions of the
ethnicity of the patients are shown in the following table.

  -------------------------- -------------------- -----------------------
  Ethnicity                  *N*                  \%

  African American           21                   10.0%

  Caucasian                  101                  48.3%

  Hispanic                   3                    1.4%

  Asian                      6                    2.9%

  Other                      7                    3.3%

  Unknown                    71                   34.0%

  Total                      209                  100.0%
  -------------------------- -------------------- -----------------------

Included were patients with potentially crossreactive substances and
medical conditions.

A total of 209 retrospective specimens were collected and tested by
FDA-approved or licensed hepatitis B assays for four HBV serological
markers (HBsAg, Anti-HBc IgM, Anti-HBc, and Anti-HBs). Characterization
based on single point specimens by the reference serological markers
demonstrated eight unique patterns:

  ---------------------------------------------------------------------------------
                     Number of  HBV                                    
                     patients   Reference                              
                                Markers                                
  ------------------ ---------- ------------ ------------ ------------ ------------
  Characterization              HBsAg        Anti-HBc IgM Anti-HBc     Anti-HBs
  based on single                                                      
  point specimens                                                      

  Acute              8          \+           --           --           --

  Acute              9          \+           +/-          \+           --

  Chronic            2          \+           --           \+           \+

  Early recovery     33         --           +/-          \+           \+

  Early recovery     17         --           --           \+           --

  HBV vaccine        32         --           --           --           \+
  response                                                             

  Not previously     107        --           --           --           --
  infected                                                             

  Uninterpretable    1          \+           --           --           \+
  ---------------------------------------------------------------------------------

Based on the above classifications the IMMULITE and IMMULITE 2000 HBsAg
results were compared to Kit A.

<table style="width:100%;">
<colgroup>
<col style="width: 32%" />
<col style="width: 0%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="9">Kit A</td>
<td colspan="3">Total</td>
<td colspan="3"></td>
</tr>
<tr class="even">
<td colspan="3" rowspan="3"></td>
<td colspan="4">+</td>
<td colspan="5">–</td>
<td colspan="3"></td>
<td></td>
</tr>
<tr class="odd">
<td colspan="9">IML HBsAg</td>
<td colspan="3"></td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="3">–</td>
<td colspan="3"></td>
<td></td>
</tr>
<tr class="odd">
<td colspan="2"><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">12</td>
<td colspan="2">5</td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="3">17</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td colspan="2"><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="3">2</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td colspan="2"><blockquote>
<p>HB Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="3">50</td>
<td colspan="3">50</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td colspan="2"><blockquote>
<p>HBV Vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="3">32</td>
<td colspan="3">32</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td colspan="2"><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">2</td>
<td colspan="3">105</td>
<td colspan="3">107</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td colspan="2"><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="3">0</td>
<td colspan="3">1</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td colspan="2"><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">14</td>
<td colspan="2">6</td>
<td colspan="2">2</td>
<td colspan="3">187</td>
<td colspan="3">209</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. Total Positive agreement is
70.0% (14/20) with a 95% CI of 45.7 to 88.1%. The Negative agreement is
98.9% (187/189) with 95% CI of 96.2 to 99.9%. Total agreement is 96.2%
(201/209) with a 95% CI of 92.6 to 98.3%. The complete data by category
is presented in the package insert.

<table>
<colgroup>
<col style="width: 32%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 14%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4">Total</td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML 2000 HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">11</td>
<td colspan="2">6</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">17</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">2</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">49</td>
<td colspan="2">50</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HBV Vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">32</td>
<td colspan="2">32</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">106</td>
<td colspan="2">107</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">13</td>
<td colspan="2">7</td>
<td colspan="2">2</td>
<td colspan="2">187</td>
<td colspan="2">209</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. The Total Positive Agreement
is 65.0% (13/20) with a 95% CI of 40.8 to 84.6%. The Negative agreement
is 98.9% (187/189) with a 95% CI of 96.2 to 99.9%. The Total agreement
is 95.7% (200/209) with a 95% CI of 92.0 to 98.0%. The complete data by
category is presented in the package insert.

**Study 3:** Specimens obtained from China were tested in the southern
United States and were comprised of 13 females and 55 males (gender for
11 patients was not reported) with an average age of 36 years, ranging
from 18 to 82 years. These were prospectively recruited patients from a
clinically well-characterized, homogeneous population: acute hepatitis B
patients who presented with symptoms typical of acute hepatitis B such
as jaundice, persistent fatigue, loss of appetite, pale stools and liver
enlargement. Their ALT and AST results were significantly elevated at
the time of diagnosis.

A total of 79 specimens were prospectively collected and tested by
FDA-approved or licensed hepatitis B assays for six HBV serological
markers (HBsAg, HBeAg, Anti-HBc IgM, Anti-HBc, Anti-HBc, and Anti-HBs).
Characterization based on single point specimens by the reference
serological markers demonstrated 11 unique patterns:

  ----------------------------------------------------------------------------------------------
                     Number of  HBV                                                   
                     patients   Reference                                             
                                Markers                                               
  ------------------ ---------- ----------- -------- ---------- ---------- ---------- ----------
  Characterization              HBsAg       HBeAg    Anti-HBc   Anti-HBc   Anti-HBe   Anti-HBs
  based on single                                    IgM                              
  point specimens                                                                     

  Acute              23         \+          --       +/-        \+         \+         --

  Acute              8          \+          +/-      \+         \+         \+         --

  Acute              1          \+          --       \+         +/-        --         --

  Acute              35         \+          \+       +/-        \+         --         --

  Acute              4          \+          \+       \+         \+         --         +/-

  Chronic            1          \+          --       --         \+         --         --

  Chronic            3          \+          +/-      --         \+         \+         --

  Chronic            1          \+          \+       +/-        \+         --         \+

  Chronic            1          \+          \+       \+         \+         \+         \+

  Recovered          1          --          --       --         +/-        --         \+

  Uninterpretable    1          \+          --       \+         \+         \+         \+
  ----------------------------------------------------------------------------------------------

Based on the above classifications the IMMULITE and IMMULITE 2000 HBsAg
results were compared to Kit A.

<table style="width:100%;">
<colgroup>
<col style="width: 32%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 14%" />
<col style="width: 0%" />
<col style="width: 14%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 11%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4"><blockquote>
<p>Total</p>
</blockquote></td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">70</td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">71</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">6</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">6</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Recovered</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">77</td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">79</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. The Total Positive agreement
is 98.7% (77/78) with a 95% CI of 93.1 to 100.0%. There was only 1
negative sample, so Negative agreement could not be calculated. The
Total agreement is 98.7% (78/79) with a 95% CI of 93.1 to 100.0% The
complete data by category is presented in the package insert.

<table>
<colgroup>
<col style="width: 28%" />
<col style="width: 0%" />
<col style="width: 11%" />
<col style="width: 0%" />
<col style="width: 15%" />
<col style="width: 0%" />
<col style="width: 15%" />
<col style="width: 0%" />
<col style="width: 15%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4"><blockquote>
<p>Total</p>
</blockquote></td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML 2000 HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>HB Acute infection</p>
</blockquote></td>
<td colspan="2">71</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">71</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HB Chronic infection</p>
</blockquote></td>
<td colspan="2">6</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">6</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Recovered</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Uninterpretable</p>
</blockquote></td>
<td colspan="2">1</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">78</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">79</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement. The Total Positive agreement
is 98.7% (78/79) with a 95% CI of 93.1 to 100.0%. The complete data by
category is presented in the package insert.

**Study 4:** Conducted in the southern United States, this study
included retrospectively collected specimens from 200 pregnant subjects.
These subjects had an average age of 28 years, ranging from 17 to 41
years.

A total of 200 specimens were tested by FDA-approved or licensed
hepatitis B assays for four HBV serological markers (HBsAg, Anti-HBc
IgM, Anti-HBc, and Anti-HBs). Characterization based on single point
specimens by the reference serological markers demonstrated three unique
patterns:

  --------------------------------------------------------------------------
  Characterization   Number of  HBV                               
  based on single    subjects   Reference                         
  point specimens               Markers                           
  ------------------ ---------- ----------- ---------- ---------- ----------
                                HBsAg       Anti-HBc   Anti-HBc   Anti-HBs
                                            IgM                   

  Early recovery     4          --          +/-        \+         \+

  Early recovery     2          --          --         \+         --

  HBV vaccine        42         --          --         --         \+
  response                                                        

  Not previously     152\*      --          --         --         --
  infected                                                        
  --------------------------------------------------------------------------

Based on the above classifications the IMMULITE and IMMULITE 2000 HBsAg
results were compared to Kit A.

<table>
<colgroup>
<col style="width: 31%" />
<col style="width: 0%" />
<col style="width: 14%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4">Total</td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">6</td>
<td colspan="2">6</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HBV vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">42</td>
<td colspan="2">42</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">148</td>
<td colspan="2">149*</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">1</td>
<td colspan="2">196</td>
<td colspan="2">197</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

\* Three specimens were not tested for IMMULITE HBsAg.

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement or Total Positive agreement.
The Total Negative agreement is 99.5% (196/197) with a 95% CI of 97.2 to
100.0%. The complete data by category is presented in the package
insert.

<table style="width:100%;">
<colgroup>
<col style="width: 34%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 12%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 13%" />
<col style="width: 0%" />
<col style="width: 11%" />
<col style="width: 0%" />
<col style="width: 0%" />
</colgroup>
<tbody>
<tr class="odd">
<td colspan="2" rowspan="4"><blockquote>
<p>Reference Serological Characterization</p>
</blockquote></td>
<td colspan="8">Kit A</td>
<td colspan="2" rowspan="4"><blockquote>
<p>Total</p>
</blockquote></td>
<td></td>
</tr>
<tr class="even">
<td colspan="4">+</td>
<td colspan="4">–</td>
<td></td>
</tr>
<tr class="odd">
<td colspan="8">IML 2000 HBsAg</td>
<td></td>
</tr>
<tr class="even">
<td colspan="2">+</td>
<td colspan="2">–</td>
<td colspan="2">+</td>
<td colspan="2">–</td>
<td></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Early recovery</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">6</td>
<td colspan="2">6</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>HBV vaccine response</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">42</td>
<td colspan="2">42</td>
<td colspan="2"></td>
</tr>
<tr class="odd">
<td><blockquote>
<p>Not previously infected</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">152</td>
<td colspan="2">152</td>
<td colspan="2"></td>
</tr>
<tr class="even">
<td><blockquote>
<p>Total</p>
</blockquote></td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">0</td>
<td colspan="2">200</td>
<td colspan="2">200</td>
<td colspan="2"></td>
</tr>
</tbody>
</table>

There was insufficient numbers for each category of subjects to
determine Negative and Positive Agreement or Total Positive agreement.
The Total Negative agreement is 100.0% (200/200) with a 95% CI of 98.2
to 100.0%. The complete data by category is presented in the package
insert.

**Study 5:** In a clinical study conducted in Europe, IMMULITE and
IMMULITE 2000 HBsAg was compared to Kit B, a commercially available
HBsAg electro-chemiluminescence immunoassay. Presented below are the
comparisons between IMMULITE HBsAg, IMMULITE 2000 HBsAg and Kit B (one
specimen per subject).

  ------------ ------------ ------------ ------------ -------------------
  Kit B                                               

  \+                        --                        

  IML HBsAg                                           

  \+           --           \+           --           Total

  412          12           26           2049         2499
  ------------ ------------ ------------ ------------ -------------------

Positive agreement = 97.2% (412/424)

95% CI = 95.1 to 98.5%

Negative agreement = 98.7% (2049/2075)

95% CI = 98.2 to 99.2%

Total agreement = 98.5% (2461/2499)

95% CI = 97.9 to 98.9%

  -------------- ------------ ------------ ------------ -------------------
  Kit B                                                 

  \+                          --                        

  IML 2000 HBsAg                                        

  \+             --           \+           --           Total

  118            12           15           2054         2199
  -------------- ------------ ------------ ------------ -------------------

Positive agreement = 90.8% (118/130)

95% CI = 84.4 to 95.1%

Negative agreement = 98.0% (2054/2096)

95% CI = 97.3 to 98.6%

Total agreement = 98.8% (2172/2199)

95% CI = 98.2 to 99.2%

**IMMULITE HBsAg Confirmatory**

In a European study, specimens with positive HBsAg results by IMMULITE
HBsAg were further tested by the IMMULITE HBsAg Confirmatory procedure.
The results are shown in the following table.

  --------------------------------- -------------------------------------
  Specimens                         HBsAg Confirmatory

  273 positive by IMMULITE HBsAg    267 confirmed positive

                                    6 not confirmed\*
  --------------------------------- -------------------------------------

\* The 6 specimens not confirmed all had extremely high counts (CPS).
Further dilution of these specimens (not done) would probably have
confirmed the positivity of these specimens.

In one clinical study in the United States, 200 specimens from
apparently healthy pregnant women with an average age of 28 years were
tested by the IMMULITE 2000 HBsAg assay, Kit A, and the IMMULITE HBsAg
Confirmatory procedure. The results are shown in the following table.

  ----------------------------------- -----------------------------------
  Specimens                           HBsAg Confirmatory

  200 negative by IMMULITE 2000 HBsAg 200 not confirmed positive
  and Kit A                           
  ----------------------------------- -----------------------------------

In two other clinical studies in the United States, test results of 19
specimens (6 females, 13 males, with an average age of 47 years) were
found to be discordant either between IMMULITE HBsAg and a commercially
available HBsAg assay (Kit A), or between IMMULITE 2000 HBsAg and Kit A.
These specimens were retested by all three assays and the IMMULITE HBsAg
Confirmatory procedure. The results are shown in the following table.

  -------------------- ----------------------- --------------------------
  Specimens            Final Determination\*   HBsAg Confirmatory

  19 specimens with    17 negative             17 not confirmed positive
  discordant HBsAg                             
  results                                      

                       2 positive              1 not confirmed positive

                                               1 confirmed positive
  -------------------- ----------------------- --------------------------

\* Negative if 2/3 or 3/3 results were negative. Positive if 2/3 or 3/3
results were positive.

In an additional study conducted in-house, 38 specimens that had tested
positive by IMMULITE HBsAg were further tested by the IMMULITE HBsAg
Confirmatory procedure and a commercially available HBsAg Confirmatory
assay (Kit A).

  -------------- ------------- ------------- ------------ -----------------
  Kit A                                                   

  \+                           --                         

  IML HBsAg                                               
  Confirmatory                                            

  \+             --            \+            --           Total

  37             0             1\*           0            38
  -------------- ------------- ------------- ------------ -----------------

Positive agreement = 100.0% (37/37)

95% CI = 90.5 to 100.0%

Negative agreement = N/A (0/1)

95% CI = N/A

Total agreement = 97.4% (37/38)

95% CI = 86.2 to 99.9%

\* This specimen had a signal reduction of 59% by IMMULITE HBsAg
Confirmatory and 48% by Kit A.

## Performance with Plasma Specimens

In a study conducted in the northwestern United States, blood specimens
from 18 chronic hepatitis B patients at different stages of the disease
were drawn into plain, heparinized, sodium citrate and EDTA vacutainer
tubes. All serum and all plasma specimens were tested by IMMULITE HBsAg
and IMMULITE 2000 HBsAg. Positive results were further tested by
IMMULITE HBsAg Confirmatory and all were conformed positive.

  ----------------- --------------- ------------------- ------------------
  Specimens         IML HBsAg       IML 2000 HBsAg      HBsAg Confirmatory
                    Results         Results             

  18 Serum          17/18 positive  17/18 positive      All 17 Confirmed

  18 Heparinized    17/18 positive  17/18 positive      All 17 Confirmed

  18 Sodium citrate 17/18 positive  17/18 positive      All 17 Confirmed

  18 EDTA           17/18 positive  17/18 positive      All 17 Confirmed
  ----------------- --------------- ------------------- ------------------

X.  **CONCLUSIONS DRAWN FROM STUDIES**

The data from both the non-clinical and clinical studies demonstrate
that acceptable performance is obtained with the IMMULITE HBsAg,
IMMULITE 2000 HBsAg and IMMULITE HBsAg Confirmatory assays to detect
hepatitis B surface antigen in human serum or plasma.

**Safety**

As a diagnostic test, the IMMULITE HBsAg, IMMULITE 2000 HBsAg and
IMMULITE HBsAg Confirmatory assays involve removal of blood from an
individual for testing purposes. The test, therefore, presents no more
safety hazard to an individual being tested than other tests where blood
is removed.

**Benefit/Risks**

The submitted clinical studies have shown that the IMMULITE HBsAg,
IMMULITE 2000 HBsAg and IMMULITE HBsAg Confirmatory assays, when
compared to reference clinical laboratory procedures, have a similar
ability to detect the presence of HBsAg in specimens from individuals
acutely and chronically infected with HBV. The rate of false positivity
and false negativity are within acceptable limits compared to the
reference assay. It has been shown that the device has no demonstrable
cross-reactivity with other viruses or organisms that may cause clinical
hepatitis. Therefore, these devices should benefit the physician in the
diagnosis of HBV.

Based on the results of the preclinical and clinical laboratory studies
the IMMULITE HBsAg, IMMULITE 2000 HBsAg and IMMULITE HBsAg Confirmatory
assays, when used according to the provided directions and in
conjunction with other serological and clinical information, should be
safe and effective and pose minimal risk to the patient due to false
test results.

XI. **PANEL RECOMMENDATION**

> Pursuant to Section 515(c)(2) of the act as amended by the Safe
> Medical Devices Act of 1990, this PMA was not the subject of an FDA
> Microbiology Devices Advisory Panel meeting because the information in
> the PMA substantially duplicated information previously reviewed by
> this Panel.

XII. **CDRH DECISION**

The applicant's manufacturing facility was found to be in compliance
with the Quality Systems Regulation (21 CFR 820). FDA issued an approval
order on July 26, 2002.

XIII. **APPROVAL SPECIFICATIONS**

Directions for Use: See labeling

Hazards to Health from Use of the Device: See Contraindications,
Warnings, Precautions and Adverse Events in the labeling**.**

Postapproval Requirements and Restrictions: See approval order.