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VIDAS Training
VIDAS HIV
Objectives
Objectives
•• know
knowthe
thebasis
basisof
ofHIV
HIVinfections
infections
•• know
know the main technicalfeatures
the main technical featuresof
ofthe
theVIDAS
VIDASreagents
reagents
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Knowthe
themain
mainsales
salesarguments
argumentsand
andtools
tools
Hepatitis profile
Every 6 seconds
someone is newly HIV infected
Every 10 seconds
someone dies from AIDS
33 millions :
the number of people living with HIV in the world today (2007 )
35 millions :
the number of people who have died of AIDS – related diseases
Epidemic
http://data.unaids.org/pub/GlobalReport/2008/JC1511_GR08_ExecutiveSummary_en.pdf
GCS - TM 0009 – 04/01/2007 – part 1 - p.4
30 % of people
do not know to be infected
Homosexuals
Homosexuels Drug
Usagersaddicts
de drogues Heterosexuals
Hétérosexuels
1200 700 700
600 600
1000
500 500
800
400 400
600
300 300
400
200 200
200 100
100
0 0 - 36% 0
94-1 96-1 98-1 '00-1 '02-1 94-1 96-1 98-1 '00-1 '02-1 94-1 96-1 98-1 '00-1 '02-1
Hepatitis profile
Why is it important
to detect the infection early?
Risk of
contamination by 30 %
contact
Primary
infection
Asymptomatic
10 %
stage
AIDS
0.1 % 0.1 %
0.01 %
0
0
Weeks Years
GCS - TM 0009 – 04/01/2007 – part 1 - p.9
Prevention of transmission
Early diagnosis
*Berrey et al., J.Infect Dis 2001
GCS - TM 0009 – 04/01/2007 – part 1 - p.10
VIDAS Training
HIV
HIV
Acquired
Immuno
Deficiency
Syndrome
GCS - TM 0009 – 04/01/2007 – part 1 - p.13
target cell
Lymphocyte T
sexual
intercourses
blood
(transfusions,
contaminated needles)
mother to child
breast milk
Risk of contamination
per contact 30 %
Primary
infection
Asymptomatic
10 %
stage
AIDS
0.1 % 0.1 %
0.01 %
0
0
Weeks Years
GCS - TM 0009 – 04/01/2007 – part 1 - p.16
50 to 70 % of primary infections
DO HAVE clinical symptoms
fever 96%
adenopathy 75%
common pharyngitis 70%
rash 70%
arthralgia 50%
diarrhea 30%
AIDS
clinical symptoms
primary
primary
infection asymptomatic
period
2 to 15 years time
AIDS
Opportunistic infections
Kaposi sarcoma
CMV
toxoplasmosis
mycobacteria infections
herpes
aspergillosis ...
no vaccine yet
antiviral therapy :
to be given as early as possible
HIV virus
Viral enveloppe:
Two layers of lipids
Spikes made of 3 gp120
and gp41 subunits
HIV matrix:
Made of p17
a lie between enveloppe and core
Viral Core :
Made of p24
p7nucleocapsid protein
Hepatitis profile
video Link
REVERSE
TRANSCRIPTASE
conversion of the viral RNA into
double-stranded viral DNA
ENZYME INTEGRASE
Integration of the viral DNA
Into the host DNA
VIRAL PROTEASE
Cleavage of longer proteins into
smaller proteins
HIV
Fourth
Generation
Third
HIV
Generation
Reactivity
RNA
Second
Generation-
p24 First
Ag
Generation
IgM
Anti- IgG
HIV Anti-HIV
0 1 2 3 4 5 6 7 8
Weeks post-infection
GCS - TM 0009 – 04/01/2007 – part 1 - p.26
HIV
1995:
HIV : Nasba HIV1 QT
History of the disease,
our story 1993: 1st HIV viral load 2004:
VIDAS DuoUltra
VIDAS P24 Ag
1991: 2 signals
1st automated The most
VIDAS HIV 1+2 test available sensisitve
1985: 1st automated 2002:
Vironostika anti test available
Real time Easy Q
HTLV III on the market Quantitative viral load
1st test
1998:
available for
screening 4th generation
1st tests
Microtiter plates
and automated
HIV bioMérieux :
The experience from screening to follow up
2 - antigen
detection
1 - antibody
detection
4.5 pg/ml
pg/ml in p24 Ag
2 - antigen
detection
1 - antibody
detection
Seroconversion panels
Clinical impact
Preventing infection
Preventing transmission
Improving patient care/ life quality
Depends Fourth
VIDAS HIV Duo on the kit Generation
Quick & Ultra
Third
HIV
Generation
Reactivity
RNA
Second
Generation-
p24 First
Ag
Generation
IgM
Anti- IgG
HIV Anti-HIV
0 1 2 3 4 5 6 7 8
Weeks post-infection
GCS - TM 0009 – 04/01/2007 – part 1 - p.47
HIV
Screening
Confirmation
Ag + Ab Antibodies only
1 2 3
Electrophoresis Transfert Sera incubation
Revelation
+ + + +
ECHANTILLONS
SAMPLES
marked Ac
- - - - Visualisation of proteins
transfert on nitrocellulose filter specifically known by
Separation of nitrocellulose + corresponding Ab
proteins transferred proteins
filter
Application to HIV
IgG only
1 WB for HIV 1 / 1 WB for HIV 2
used for specificity
limited in sensitivity : less than ELISA techniques
variations between - suppliers
- batchs from a same supplier
the lab has to «validate» each lot before use
Poor reagents stability
Interferences : blood, bilirubin, auto-immune diseases...
Necessity to use controls for interpretation
Subjective interpretation : visual
the interpretation depends on the local registration
No detectable after
during asymptomatic period, P24 is complexed with anti P24
60 tests
90 mins
same protocol as VIDAS HIV P24 II
test code : P24
VIDAS reaction
conjugate
anti P24
Ag P24 E +
monoclonal *
Confirmation reaction :
1 - neutralization 2 - VIDAS reaction
Ag P24
4
-
P2
250 µl sample
Procedure
30 min
37 °C
200 µl treated
sample
P24 II Ag strip
This algorithm is proposed as an indication and is valid for the majority of cases. Please take into consideration the recommendations in force in your country.
(1) If clinical signs or risk factors require a new bleeding two weeks after, it is also possible to ask for a viral load on a 2nd immediate bleeding to provide the
patient with a more rapid diagnosis orientation.
GCS - TM 0009 – 04/01/2007 – part 1 - p.59