HIV Diagnostic Tests
Transcript of HIV Diagnostic Tests
HIV Diagnostic Tests
Bernard M. Branson, M.D. Associate Director for Laboratory Diagnostics
Division of H IV/AIDS Prevention Centers for Disease Control & Prevention
The views expressed in this presentation are those of the author and do not necessarily represent those of the Centers for Disease Control and Prevention
Generations
1st and 2nd Generation EIA Plasma/serum (1 h/37o C)
Plasma/serum
Color reagent
Antigen 1st - Viral lysate 2nd – Recombinant proteins or synthetic peptides
IgG HIV antibody
enzyme
anti-human IgG
Detects HIV IgG
Enzyme- detection
EIA
Western blot
IFA
3rd Generation “Sandwich” EIA
Antigen: Recombinant proteins or synthetic peptides Enzyme-
detection
Color reagent
Plasma/serum HIV antibody
enzyme
HIV antigen
Detects HIV IgM or IgG
IgG IgM
4th Generation Combo EIA
Color reagent
Enzyme- detection
HIV antigen p24 antibody
Plasma/serum HIV antibodies p24 antigen
HIV antigen
p24 antibody
Detects IgM or IgG
antibody or p24 antigen
i
HIV Infection and Laboratory Markers
HIV RNA (plasma) HIV Antibody
0 10 20 30 40 50 60 70 80 90 100
HIV p24 Ag
22
1st gen
2nd gen
3rd gen
Days
Modified after Busch et al. Am J Med. 1997
Infection
9
4th gen
1985 Abbott
HIV-1 EIA
1987 Vironostika
EIA
1992 Abbott
HIV-1/HIV-2 EIA
2000 Genetic Systems
HIV-1/HIV-2 Peptide EIA
2002 OraQuick
HIV-1/HIV-2 Rapid Test
2003 GS HIV-1
HIV-2 Plus O EIA
2004 Multispot
HIV-1/HIV-2 Rapid Test
2006 Aptima
Qualitative RNA
2006 Advia Centaur
1/O/2 CIA
2008 Ortho Vitros
HIV 1+2 CIA
1991 Cambridge
Western blot
1992 Fluorognost
IFA
1998 Genetic Systems
rLAV (HIV-1)
2003 Unigold Reveal
HIV-1 Rapid Tests
2010 Abbott
Architect Ag/Ab
Combo CIA
2011 Bio-Rad Ag/Ab Combo
EIA
2010 INSTI HIV-1
Rapid Test
2009 Avioq HIV-1
EIA
3rd & 4th gen lab screening tests
2nd gen rapid tests 1st gen confirmatory
Uni-Gold Recombigen 2003
Multispot HIV-1/HIV-2 2004
Reveal G3 2003
OraQuick Advance 2002
Clearview Complete HIV 1/2 2006
Clearview HIV 1/2 Stat Pack 2006
Rapid HIV Test Results without
Rapid Test Kits
On-board Refrigeration of Multiple Different Assays
Random Access Multiplatform analyzers for HIV testing
STAT sample requests without pausing Results in < 60 minutes
Random Access Multiplatform analyzers for HIV testing
ADVIA® Centaur™ H IV 1/O /2 Enhanced (EH IV) Chemiluminescent
immunoassay
3rd generation format - H IV-1: gp41, p24 - H IV-2: gp36 - group O
Time to result < 1 hour
FDA-approved July 2006
Ortho VITROS ECi/ECiQ Chemiluminescent
immunoassay
3rd generation format - H IV-1: gp41, gp120, p24 - H IV-2: gp36 - group O
Time to result < 1 hour
Repeat only borderline results
FDA-approved March 2008
Chemiluminescent immunoassay
Detects p24 antigen and H IV antibody
Time to result: 29 minutes
FDA-approved
June 22, 2010
Abbott Architect 4th Generation Ag/Ab Combo Assay
APTIMA Qualitative HIV-1 RNA Assay Aid to H IV-1 diagnosis
Diagnosis of acute H IV-1
infection in antibody-negative persons
Confirmation of H IV-1 infection in antibody-positive persons when it is reactive
FDA-approved July 2006
Bio-Rad GS HIV Combo Ag/Ab EIA
Microwell plate EIA
3rd generation format: - HIV-1: gp160 - HIV-2: gp36 - Group O
p24 antigen
FDA-approved July 25, 2011
Relative Seroconversion Sensitivity
0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
-50 -40 -30 -20 -10 0 10 20 30
Genprobe
Architect 4th
BioRad 4th
Advia 3rd
Vitros 3rd
Abbott 3rd
BioRad 3rd
Insti
Multispot
Reveal
Statpak
Complete
Unigold
Oraq Adv
Cum
ulat
ive
frequ
ency
of p
ositi
ve te
sts
days to WB positive
26 seroconverters were analyzed with 14 tests 17 seroconverters with WB positive used for cumulative frequency analysis
Sequence of Test Positivity Relative to WB 166 specimens, 17 Seroconverters - 50 % Positive Cumulative Frequency
Modified from Masciotra et al, J Clin Virol 2011
Days before WB positive
WB
posi
tive
APT
IMA
(-26)
GS
1/2+
O (
-12)
Mul
ti-Sp
ot (
-7)
Reve
al G
3,
(-6)
Viro
nost
ka (+
2)
Ora
Qui
ck (-
1)
Uni
gold
(-2)
25
20
10
5 0 15
Arch
itect
Ag/
Ab C
ombo
(-2
0)
COM
PLET
E H
IV-1
/2 (
-5)
HIV
-1/2
STA
T-PA
K (-
5)
Advi
a (-
14)
Vitr
os (
-13)
INST
I (-9
)
Bio-
Rad
Ag/
Ab C
ombo
(-1
9)
and Owen et al, J Clin Micro 2008
Avio
q
Det
erm
ine
Ag/A
b Co
mbo
(-1
5)
Acute Infections in MSM detected by RNA only
0.3% of 14,005 frequently tested MSM in Seattle STD clinic; 20% of all HIV infections detected
26 (74%) of 35 AHI cases detected in LA at MSM clinic; 25% of all HIV infections detected
0.08% of 21,425 STD clinic patients in New York City; 9% of all HIV infections detected; 94% were MSM
- Stekler et al, Clin Infect Dis 2009
- Shepard et al, MMWR 2009
- Patel et al, Archives Int Med 2010
RNA vs. 4th Generation Ag/Ab Assay
RNA+ / 3rd gen-negative specimens detected by 4th generation EIA: 38 of 46 (83%) – Australia* 10 of 14 (71%) – CDC AHI study** 51 of 61 (84%) – CDC panel***
4 days after RNA – 9 seroconversion panels***
* Cunningham P, HIV Diagnostics Conf 2007 ** Patel P, CROI 2009 *** Owen M, CROI 2009
On the Horizon…
Ag/Ab Rapid Test
Point-of-careHIV-1/2 antibody differentiation assay with
automated reader
Point-of-care finger-stick viral load and CD4
analyzer