FibroTest-ActiTest

FibroMAX
New Non-Invasive Liver Disease BioMarkers
THE ONLY TESTS FOR THE DIAGNOSIS
AND STAGING OF LIVER FIBROSIS
AND THE MOST COMMON LIVER DISEASES
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In order to best manage the patient, it is important to recognize the natural history of
viral hepatitis : characterized by brosis progression and necro-inammatory activity.
Activity and fbrosis are two major histological features of chronic viral hepatitis, and in
several cases associated with steatosis including hep C-genotype 3. The METAVIR system
(one of the validated scoring systems) assesses histological lesions in chronic
hepatitis by activity and fbrosis.
The fbrosis stage is correlated to the progression to cirrhosis of the patient
and its propensity eg a HCV infected patient:
The major histological features of viral hepatitis
activity and brosis are essential for providing
information about the condition of the liver
HCV Infection: A virologic and fibrotic disease
Activity grade predicts the level of necrosis. But of note fbrosis alone
is the best marker of ongoing fbrogenesis. While in the majority of patients,
fbrosis and activity are correlated, in about 1/3 of patients signifcant activity
is not necessarily a marker of severe disease in its own right.
It is the combination of factors, as well as age, gender and certain risk factors
that determine the appropriate diagnosis and stage of liver disease.
It is important to regularly
monitor the major histological features
of hepatitis activity and brosis.

20% acute
Hemorrhage
80% chronic
F0
F1
F2
F3
F4 (=cirrhosis)
Hepatic
Insufency
Cancer
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
As such,
numerous studies strongly suggest that due
to the limitations and risks of biopsy, as well as the
improvement of the diagnostic accuracy of biochemical
markers, liver biopsy should no longer be considered
mandatory (1).
Only 5%
of patients at risk
of brosis
have a liver biopsy
Traditionally, liver biopsy has been utilized to assess the histological features
of the liver and hence estimate the degree of liver damage.
However, this procedure has several drawbacks :
it is invasive
it is prone to complications, ranging from minor (up to 30% experience pain)
through to more severe (including death in approximately 0.03%)
there is considerable sampling variability (up to 40% for fbrosis staging)
there is a high intra and inter-pathologist variability
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
Liver biopsy is undesirable to patients
& associated with serious complications
The BioPredictive tests:
FibroTest-ActiTest

is the combination of FibroTest

& ActiTest

;
FibroMAX

is the combination of up to ve non-invasive
liver tests: FibroTest

, ActiTest

, SteatoTest, NashTest

and AshTest

.
FibroTest-ActiTest

and FibroMAX

:
the innovative diagnostic and staging tools
for the most common liver diseases
FibroTest-ActiTest

> FibroTest

: diagnoses hepatic fbrosis

> ActiTest

: assesses viral necro-infammatory activity

FibroMAX

> FibroTest

: diagnoses hepatic fbrosis

> SteatoTest

: diagnoses hepatic steatosis (otherwise known as fatty liver)

the most common cause of ALT and GGT abnormalities
> ActiTest

: assesses viral necro-infammatory activity

> AshTest

: diagnoses severe alcoholic steatohepatitis (ASH) in excessive drinkers

> NashTest

: diagnoses non-alcoholic steatohepatitis (NASH)

in patients who are overweight, insulin resistant,
have diabetes or hyperlipidemia
The BioPredictive tests work by using an algorithm based on the patients sex, age,
weight, height and specifc blood biomarkers*.
*see page 8 for full list of specic blood markers.
In Situ :
Liver Injury
Fibrotic Matrix
Activated Stellate Cells
In Serum :
FibroTest-ActiTest
Gender, Age
Alpha2Macroglobulin
Total Bilirubin
Gamma GT
Apolipoprotein A1
Haptoglobin
ALT
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FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
The BioPredictive Patented New Technology...
Chronic Hepatitis
C or B
BioPredictive test required and result sample
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Viral, Metabolic
or Alcoholic
Hepatitis when
symptoms
or risk factors
are difcult
to decifer
> FibroTest-ActiTest

F
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A
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> FibroMAX

Patient Prole
F4
F3
F2
F1
F0
FibroTest Actitest
A3
A2
A1
A0
Score 0.50 Score 0.38
(F2) (A1-A2)
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
F4
F3
F2
F1
F0
FibroTest Steatotest ActiTest AshTest NashTest
S3
S2
S1
S0
Score 0.50 Score 0.49 Score 0.38 Score 0.03 Score 0.75
(F2) (S1-A2) (A1-A2) (H0) (Nash)
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
H3
H2
H1
H0
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
N2
N1
N0
A3
A2
A1
A0
The Biopredictive tests
provide accurate results instantly
FibroTest

Fibrosis
staging by
METAVIR
system
ActiTest

Activity by
METAVIR
system
SteatoTest

Steatosis
severity score
AshTest

Alcoholic
steato-hepatisis
score
NashTest

Metabolic
non-alcoholic
steato-hepatisis
score
F0 =
no brosis
A0 =
no histologic
activity
S0 =
no steatosis
H1 =
minimal ASH
N0 =
no NASH
F1 =
portal brosis
without septa
A1 =
minimal activ-
ity
S1 =
minimal
steatosis,
<5%
hepatocytes
with steatosis
H2 =
moderate
ASH
N1 =
borderline
NASH
F2 =
portal brosis
with few
septa
A2 =
moderate
activity
S2 =
moderate
steatosis,
6%-32%
hepatocytes
with steatosis
H3 =
severe ASH
N2 =
NASH
F3 =
numerous
septa without
cirrhosis
A3 =
severe activity
S3 S4 =
severe
steatosis
33% - 100%
hepatocytes
with steatosis
F4 =
cirrhosis
The BioPredictive test scores
are classied as follows :
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
BioPredictive test required and result sample
These algorithms have been scientifcally validated and quality-monitored to ensure that
the results delivered are of the highest analytical standards (assays are performed using
methods which have been standardized with respect to reference methods and/or
reference materials). The algorithms have been patented.
The FibroTest-ActiTest

results are interpreted

as follows
*
:
FibroTest METAVIR
Fibrosis
Stage
Estimate
Kendall
Fibrosis
Stage
Estimate
Ishak
Fibrosis
Stage
Estimate
0.75-1.00 F4 F4 F6
0.73-0.74 F3 - F4 F3 - F4 F5
0.59-0.72 F3 F3 F4
0.49-0.58 F2 F1 - F3 F3
0.32-0.48 F1 - F2 F1 - F3 F2 - F3
0.28-0.31 F1 F1 F2
0.22-0.27 F0 - F1 F0 - F1 F1
0.00-0.21 F0 F0 F0
F4
F3
F2
F1
F0
Score 0.33 (F1-F2)
> Cirrhosis
> Numerous septa
without Fibrosis
> Portal Fibrosis
with few septa
> Portal Fibrosis without
septa
> No Fibrosis
Precautions for Use
The reliability of results is bound with the respect of the preanalytical and analytical
conditions recommended by BioPredictive.
The Tests have to be deferred for: acute hemolysis, acute hepatitis, acute infammation,
extra hepatic cholestasis.
The advice of a specialist should be sought for interpretation in chronic hemolysis
and Gilberts syndrome.
The Test interpretation is not validated in liver transplant patients.
Isolated extreme values of one of the components should lead to caution in interpreting the results.
In case of discordance between a biopsy result and a Test, it is advised to seek for the advice
of a specialist. The causes of these discordances could be due to a faw of the Test or to a faw
in the biopsy: i.e. a liver biopsy has a 33% variability rate for one fbrosis stage.
FibroTest and SteatoTest are interpretable for chronic hepatitis B and C, alcoholic
and non alcoholic steatosis.
NashTest is interpretable for non-alcoholic steatosis.
AshTest is interpretable for alcoholic steatosis.
ActiTest is interpretable for chronic hepatitis B and C.
The validation and analytical standardization of FibroTest-ActiTest

and FibroMAX

have been established in several clinical studies. There are over 40 publications
available that review and assess the tests and these can be found out at:
> Fibrosis stage
*sample test patient
> Patients
test score
FibroTest
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
www.biopredictive.com
How to perform the BioPredictive test
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
> Log onto www.biopredictive.com to register. REGISTRATION IS FREE.
Registration will provide you with your username and password
> The website will then direct you to the appropriate page to enter your username
and password, and then you will need to follow the steps below :
Click on Connection to validate
Fill in the FibroTest-ActiTest

form (remember to never enter the patient name only

their internal number in the Internal Reference feld to ensure full privacy)
Click on Save to validate
The summary of values you entered will then re-appear. You will need to either :
(1) Click on Modify if needed or,
(2) Continue and Click on Validate to defnitely generate fnal result
If you choose a :
(a) credit card payment, enter your card number on the secure site PAYBOX.
Click on Retour Boutique to obtain the test result
(b) If you choose another payment method, test result appears automatically
> Ask for a color printout by clicking on Print the information
> You will receive an invoice by e-mail regardless of your payment method
> If you would like to perform a new test, simply click on Request a new test
Be sure to keep your user name and password in a safe place
for subsequent log-ons.
FibroTest METAVIR
Fibrosis
Stage
Estimate
Kendall
Fibrosis
Stage
Estimate
Ishak
Fibrosis
Stage
Estimate
0.75-1.00 F4 F4 F6
0.73-0.74 F3 - F4 F3 - F4 F5
0.59-0.72 F3 F3 F4
0.49-0.58 F2 F1 - F3 F3
0.32-0.48 F1 - F2 F1 - F3 F2 - F3
0.28-0.31 F1 F1 F2
0.22-0.27 F0 - F1 F0 - F1 F1
0.00-0.21 F0 F0 F0
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
FibroTest-ActiTest

and FibroMAX

can be repeated as necessary, according
to the patients prole and disease severity.

Technical recommendations result from several published studies co-ordinated by

the Biochemistry Department of Piti-Salptrire Hospital Group in Paris.
These studies assessed the intra-laboratory and the inter-laboratory variabilities
and sera storage. Test result reliability depends on the rigorous usage
of the technical recommendations. Conversely, failing to follow these technical
recommendations could signifcantly impair the diagnostic value
of the BioPredictive tests.
The following recommendations are specically related to :
1. the blood sample itself
2. the analytical method and the reagents
3. calibration, and controls
4. the criteria for reliability of the assays
The different biochemical tests :
Technical Recommendations
FibroTest-ActiTest

alpha-2 macroglobulin
haptoglobulin
apolipoprotein A1
total bilirubin
GGT
ALT
FibroMAX

alpha-2 macroglobulin
haptoglobin
apolipoprotein A1
total bilirubin
ALT
AST
GGT
fasting glucose
triglycerides
total cholesterol
A. Pre-analytical phase: essential conditions required
Blood sample
Blood is obtained by venipuncture (the classical requested 12 hour patient fast
is necessary for FibroMax but is not required for Fibro Test-ActitTest

)
Blood samples are collected in 5-ml or 7-ml tubes, without anticoagulant
Blood sample is centrifugated within 2 hours of the blood collection
The centrifugation conditions (speed and time) must be in conformity
with recommendations of tube manufacturer
Storage and transport conditions, pretreatment before analysis
Biochemical assays are usually performed on fresh serum
Serum can be decanted and stored for no more than 72 hours at + 2C/ + 4C,
while protected from light to avoid bilirubin degradation
The assays of the specifc proteins can be carried out on serum stored
at +2C / +4C for no more than 5 days
If you intend to defer analysis beyond the recommended time,
serum must be frozen at 80C from the start
Freezing and thawing can be done only once
Interference
Lipids and hemolysis interfere with measurements. It is always possible to dilute a
lightly lipemic serum according to the recommendations of the standardized methods.
Hemolysed and/or hyperlipemic serum must be rejected.
B. BioPredictive Test parameter assays
The performances of the assay methods of the FibroTest and ActiTest
have been verifed on analyzers and under the conditions specifed in
the Tecnical Recommendations document which can be sourced from
www.biopredictive.com. They are a guarantee of the quality of the results
of the BioPredictive tests.
The materials of assessment (including the reagents and analyzers), as well
as the various assay methods (ie immunonephelemetric and nephlometry for protein
assays) are also covered in detail in the Technical Recommendations document.
For further information on technical recommendations, please visit
www.biopredictive.com. After reviewing the website information, if you still have
questions with regard to the compatibility of yours analyzers or reagents,
please contact BioPredictive directly.
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
The BioPredictive Tests are simple,
convenient and cost-effective
About BioPredictive
BioPredictive

is a biotechnology company based in Paris, France.

The aim of BioPredictive

is the discovery and development of new generation

of diagnostic tests employing non-invasive techniques designed to facilitate
the care and treatment of patients.
For more information please visit
www.biopredictive.com
References :
(1) Poynard T et al.: Overview of diagnostic value of biochemical markers of liver brosis (FibroTest, HCV
FibroSure) and necrosis (ActiTest) in patients with chronic hepatitis C. Comparative Hepatology 2004; 3: 8.
(2) Ratziu V et al: Screening for advanced brosis using non-invasive biomarkers (FibroTest- FIBROSURET)
in patients consulting for hyperlipidemia. Aliment Pharmacol Ther. 2006 (Online Accepted).
(3) S. Jacqueminet et al: Efcacy of screening for advanced brosis (AF) using non-invasive biomarkers
(FibroTest-FibroSURE) (FT) in diabetic patients (DI)
FibroTest-FibroSURE in Diabetic Patients: J. Hepatol. 2006; 44: S260.
Imbert-Bismut F et Al.: Biochemical markers of liver brosis in patients with hepatitis C virus infection :
a prospective study, Lancet 2001: 357: 1069-75.
Thierry Poynard, Hepatits C and B: Management and Treatment.
Second Addition. Published by the Taylor and Francis Ed 2004.
Poynard T et al; The diagnostic value of biomarkers (SteatoTest) for the prediction of liver steatosis.
Comp Hepatol. 2005; 4: 10.
Poynard T et al; Diagnostic value of biochemical markers (NashTest) for the prediction of non alcoholo
steato hepatitis in patients with non-alcoholic fatty liver disease. BMC Gastroenterol. 2006; 6: 34.
Thabut D et al.; The diagnostic value of biomarkers (AshTest) for the prediction of alcoholic steato-hepatitis
in patients with chronic alcoholic liver disease. J Hepatol. 2006; 44: 1175-85.
FibroTest-ActiTest & FibroMAX New Non-Invasive Liver Disease BioMarkers
Step 1
Physician prescribes
FibroTest-ActiTest
or FibroMAX
Patient has blood
sample taken at
local biomedical
laboratory
Data from blood
test results are
entered directly
online at
www.
biopredictive.com
by biologist and
results are generated
immediately
@
Biologist provides
report back
to physician
1
2
3
4
Step 2 Step 3 Step 4
F4
F3
F2
F1
F0
FibroTest Steatotest ActiTest AshTest NashTest
S3
S2
S1
S0
Score 0.50 Score 0.49 Score 0.38 Score 0.03 Score 0,75
(F2) (S1-A2) (A1-A2) (H0) (Nash)
A3
A2
A1
A0
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
H3
H2
H1
H0
1.00 _
0.75 _
0.50 _
0.25 _
0.00 _
1,00 _
0,75 _
0,50 _
0,25 _
0,00 _
N2
N1
N0
40, rue du Bac
75007 Paris, France
contact@biopredictive.com
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