LIAISON XL Achieving Blood Safety through CLIA …laborexpert.hu/dat/pdf/a_cucchietti.pdf ·...

“Serological Testing for Transfusion Transmissible

Viruses: a LIAISON® XL overview”

A. Cucchietti

Senior Corporate Product Manager Hepatitis and Retrovirus

LIAISON® XL – Achieving Blood Safety through

CLIA Technology

Workshop Laborexpert

May 16th, 2013 – Siófok, Hungary

• Headquartered in Italy, DiaSorin is a global leader in the field of

biotechnologies.

• For over 40 years the company has been developing, producing and

introducing to the market reagent kits for in vitro diagnostics worldwide.

• Its line of products can meet the needs of the following clinical

areas: infectious diseases, hepatitis and retrovirus, cardiac markers, bone

metabolism (i.e. Vitamin D), oncology and endocrinology.

Today, DiaSorin can offer to the market an assays’ menu that is unique for its

width and for its exceptional number of specialty tests, which identify DiaSorin

as " the" in vitro diagnostics specialist.

DiaSorin, THE DIAGNOSTIC SPECIALIST

1

INNOVATION UNIQUENESS

CLIENT FOCUS EXCLUSIVITY

WIDE-RANGING DISTRIBUTION SYSTEM

FINANCIAL STRENGTH

WS Hungary, May 2013

2

DiaSorin worldwide presence


OUR HISTORY > 40 YEARS OF VALUE

Continuous innovation, top-quality products and long-standing commitment to

clients needs

3 WS Hungary, May 2013

LIAISON XL, THE NEW FRONT LINE ANALYZER

Compact and Efficient Design

LIAISON reagents Quality

Wide range of Assays (Specialties)

Easy to use & Reliable

All process in control

Real continuos loading of samples

and reagents

New Technical solutions to achieve

safety quality results:

Single cavity reaction & Sample Tips

to prevent carry over

Sample detection by pressure sensor


LIAISON FAMILY INSTALLED BASE

> 4.700 units installed worldwide


78-82 89 90 91 92 93 94 95 96 97 98 99 00 01 02 03 04 05 06 2010

1° HIV assay & HCV IP filing

Murex acquires

Wellcome

HIV HTLV

Syphilis

Abbott acquires Murex

HCV v 4.0 HBsAg,

other HBV assays

HIV Ag/Ab HAV IgM

Anti-HBc HAV Total

HDV

HCV Type

HCV Ag/Ab

DiaSorin acquires Murex

DiaSorin RIA & ELISA HBV

(FDA)

DiaSorin ELISA HCV HIV

DiaSorin ELISA HIV

tetra

DiaSorin LIAISON HBV

and HAV

DiaSorin Launches

LIAISON XL HCV, HIV

HBsAg

Know-how and reputation of Murex in the

field of Hepatitis and HIV will allow DiaSorin

to leverage a strong brand.

The added value of the two brands.

HBV assays

DiaSorin history: Murex and DiaSorin


DiaSorin Murex® in the world

More than 700 Murex users, in 80 different countries

*based on sales of Murex assays in the last 6 months


DiaSorin: a global presence DiaSorin LIAISON® XL in the world

Already 674 instruments installed in the world.

More than 100 users of HBsAg, HIV, HCV and HTLV, out of

which, 20 are blood banks.

Are blood banks

2 LATAM

312 Europe

122 APAC Region

110 US & Canada

121 Italy

Installed Base (February 2013)

8

7 Russia


CHEMILUMINESCENCE IMMUNOASSAY (CLIA) FOR

THE QUANTITATIVE DETERMINATION OF

SURFACE ANTIGEN OF HBV

IN HUMAN SERUM OR PLASMA SAMPLES

The LIAISON XL HBsAg Quant assay is intended to be used: in the diagnosis of HBV infection

in a HBV screening test for donated blood

and plasma

Latest pieces of evidence support the role of

HBsAg as a predictive marker for the anti-HBV

treatment response (i.e. PEG Interferone)

LIAISON® XL murex HBsAg Quant


Magnetic particles coated

with antibodies to HBsAg

(mouse monoclonal)

+ HBsAg in

samples, and

controls

Mouse monoclonal IgG to

HBsAg having balanced

reactivity for ad and ay

subtypes, conjugated to an

isoluminol derivative

STEP 1

STEP 2

HBsAg binds to

the solid phase

Antibody conjugate reacts

with HBsAg already bound

to the solid phase

Sandwich 2 steps

Incubation

(21’ - 66 cycles) Wash with

Wash/System

liquid.

Incubation

(31’ - 99 cycles)

Wash with

Wash/System

liquid.

Add the Starter Reagents

and measure the light

emitted

150µL sample

30µL Assay Buffer

180µL Tracer

20µL Assay Buffer



HBV Mutant: what is the issue related to?

Potential impact on the blood supply


New model: HBsAg MoAbs on 3 targets

IgG ID Use a.a. epitope HBs region

5C3 Solid Phase124-147

conformational“a loop”

121 Solid Phase151-170

linearTransmembrane

163 Solid Phase31-50 linear

Inner hydrophilic

124 Conjugate124-147





linear

Surface

(Mainly

conserved)

111 Conjugate61-69linear

Inner hydrophilic

Harsh

conditions

…LIAISON® XL murex HBsAg Quant

3 MoAbs

4 MoAbs

Targets


Mutatíon Panel

ID

TARGET REGION - Mutant Panel

Panel ID 3,7,8,9,10 => “a” loop (124-147)

Panel ID 1,2,4,5 => Surface (110-123)

Panel ID 6 => Combined

DiaSorin’s proof : A Recombinant HBsAg panel


HBsAg Mutations..influence the results of assays

DiaSorin

Rec Mutants' Panel

DiaSorin

LIAISON XL

murex HBsAg

Quant ≥ 0,05

IU/mL

Abbott

ARCHITECT

HBsAg

Quant ≥ 0,05

IU/mL

Abbott

ARCHITECT

HBsAg II

Qual ≥ 1

Abbott

AxSym

HBsAg

Qual ≥ 2

Abbott

PRISM

HBsAg

Qual ≥ 1

Siemens

Centaur

HBsAg II

Qual ≥ 1

Roche Cobas E600

Elecsys

HBsAg II

Qual ≥ 1

Beckman

UniCel Dxi800

HBsAg

Qual ≥ 1

Biomerieux

VIDAS HBsAg

Qual ≥ 1

Ortho Vitros

3600 HBsAg

Qual ≥ 1

HBsAgQ11_Negative

G145R

D144A

P142L-G145R

P142S-G145R

T123N

122+DT

122+DT-G145R

I110R-S117I-G119R-T123N

T123N-T124S

P142L-F/Y143H-D144E-G145R

Reactive 10 8 9 4 4 7 5 5 7 5

Non Reactive 1 3 3 7 7 4 6 6 4 6

% Positive 100% 80% 90% 40% 40% 70% 50% 50% 70% 50%

Ranking 1 3 2 9 9 4 6 6 4 6


negative

positive

Source of the data

published

Di Natale, AMCLI

2011; PEI ISBT

2012

Adler,

Hepatitis

2012

Internal

data

Internal

data

Dorizzi,

AACC

2012

Dorizzi,

AACC 2012

Caprini,

ESCV

2012

Dorizzi,

AACC

2012

Dorizzi,

AACC

2012

LIAISON® XL murex

HBsAg Quant assay

achieved the best

score for mutant

detection even at

lower concentrations.

Poster 22nd Regional Congress of the ISBT, Taipei, November 19 - 23, 2011.

Mutations in the HBsAg influence the results of

HBsAg assays

Evaluation of analytical sensitivity and HBsAg genotype and mutant detection

capability of 5 commercially available assays.


Mutations in the HBsAg influence the results of

HBsAg assays

PEI evaluation of LSN XL HBsAg Quant


LIAISON® XL murex HBsAg Quant assay

showed excellent mutant detection

sensitivity, seroconversion and

genotype/subtypes sensitivity

18

PEI evaluation of LSN XL HBsAg Quant

Has been certified by PEI also:

• LIAISON XL murex Ab/Ag

• LIAISON XL HCV Ab

• LIAISON anti-HBc


Products with unique features

• Only one parameter for HBV screening (qual) and monitoring (quant) • Easy of use

• Better patient follow-up

• Cost reduction

• Enhanced mutants detection due to an unique assay design • Monoclonal Abs towards internal, external and transmembrane

(“conserved”) regions to guarantee 100% mutants detection

• High specificity (PI Blood Donors 99.98%)

• Earlier detection proven by seroconversion data

• Excellent correlation with 2nd IS WHO

• Automated dilution for sample out of assay range


CHEMILUMINESCENCE IMMUNOASSAY (CLIA) FOR THE COMBINED

QUALITATIVE DETERMINATION OF

P24 ANTIGEN OF HIV AND ANTIBODIES TO HIV-1 (GROUP M AND

GROUP O) AND/OR HIV-2


The LIAISON XL HIV Ag/Ab assay is intended to be used:

in the diagnosis of HIV1/HIV2 infection

as a HIV screening test for donated blood and plasma

LIAISON® XL murex HIV Ab/Ag


Solid phase and conjugates tailor made for reliable results by using

unique selected Monoclonal Antibodies (Mabs) to HIV p24 antigen and

soluble and properly folded recombinant antigens: HIV-1 gp41 (group

M and group O) as well as HIV-2 gp35 (both obtained in E. Coli)

Core technology

18



18



Distinct Ag/Ab signals may contribute to early

recognition of seroconversion.

Source: Pronier, ESCV 2012

+

Recombinant HIV-1 gp41 (group

M and group O) as well as HIV-2

gp35 recombinant antigens

conjugated to an isoluminol

derivative

STEP 1

STEP 2

Sandwich 2 steps

Magnetic particles coated with rec HIV-1 gp41

(group M and group O) as well as HIV-2 gp35

recombinant antigens (both obtained in E. coli)

specimen diluent

Spec. antibodies to HIV 1 (group

M and group O) and/or to HIV-2 in

human serum or plasma samples

100uL

Incubation

(12’ 36” - 36 cycles)

+

Wash with

Wash/System

liquid.

Incubation

(33’ 36”- 96 cycles)

conjugate diluent

Wash with

Wash/System liquid



emitted

LIAISON® XL murex HIV Ab/Ag – Test Ab Format


+

p24 antigen of human immunodeficiency

virus type 1 (HIV-1) Sample

100 uL

IgG (monoclonal) to HIV p24

antigen, conjugated to an

isoluminol derivative

Add the Starter

Reagents and

measure the

light emitted

Sandwich 1 step

Biotinylated antibodies

(monoclonal) to HIV p24

antigen

Incubation

(12’ 36” - 36 cycles)

+

LIAISON® XL murex HIV Ab/Ag– Test Ag Format



• Combined qualitative determination of p24 Ag of HIV-1 and

antibodies to HIV-1 (group M and group O) and/or HIV-2 (4th

generation assay)

• One patient result from the combination of each test result

(available in the print out) • Better patient follow-up

• Cost reduction

• Good sensitivity on seroconversion data (Dr Ly, ESCV 2011)

• Good sensitivity on HIV-2 samples (Dr Ly, ESCV 2011)

• High specificity (PI Blood Donors 99.7%)

21



CHEMILUMINESCENCE IMMUNOASSAY (CLIA) FOR THE

QUALITATIVE DETERMINATION OF

ANTIBODIES TO HCV


The LIAISON XL HCV Ab assay is intended to be used:

in clinical HCV settings

in HCV screening test for donated blood and

plasma

LIAISON® XL murex HCV Ab


+

STEP 1

STEP 2

Indirect 2 steps

Incubation

(21’ - 66 cycles)

+

Wash with

Wash/System

liquid.

Incubation

(10’- 36 cycles)

Wash with

Wash/System liquid



emitted

25µL Sample

150µL diluent

150µL Tracer

NS4

NS4 p22

p22

MP coated with HCV core

and NS4 rec Ag and

streptavidin-coated MP

C33

C33 25µL Antigen

23



In a total of 678 HCV Ab(+) samples tested, 294 were characterized by genotype,

50 were from chronic phase patients and 20 from acute phase patients.

100% sensitivity

Number of samples for each genotype

88

4035

94

22

15

1

0

10

20

30

40

50

60

70

80

90

100

1 2 3 4 4 non-a 5 6

genotype

nr

Genotype distribution

Japan

South Africa

Egypt

Recognition of different genotypes, even in the

case of highly divergent variants…

25



Excellent seroconversion sensitivity for early HCV infection

26



• Qualitative determination of specific antibodies to hepatitis C,

using recombinant antigens to maximise specificity and

sensitivity.

• Excellent agreement with Ortho HCV and Architect HCV assays.

• Excellent sensitivity on different genotypes and stages of infections.

• Good sensitivity on seroconversion data.

• High specificity (PI Blood Donors 99.7%).

21




THE LIAISON FAMILY COLLECTION



MERCI

TACK SÅ MYCKET

MUCHAS GRACIAS

OBRIGADO

感謝=

THANKS

GRAZIE

DANKE

xièxie ANGE TACK

KIITOS

DEKUJI

terima kasih

ขอบคุณ

धन्यवाद

تشکر از دیگران

شكرا جزیال

شكرا جزیال

29

спасибо

köszönöm

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