Module 3:

Overview of HIV Testing

Technologies
2
Learning Objectives
At the end of this module, you will be able to:

Discuss settings where HIV testing will be part of

service delivery during an era of epanded
services

Discuss the spectrum of testing technologies for

HIV

!plain the advantages and disadvantages of

HIV rapid tests

Accurately recogni"e individual test result as

reactive, non#reactive, or invalid
Lab workers Health workers Counselors
$
Content Overview

!pansion of HIV rapid testing

%pectrum of HIV diagnostic tests

&hallenges with HIV testing

%pectrum of HIV testing technologies

Advantages and disadvantages of HIV

rapid testing

'hree formats of rapid tests

(eading individual test results

Lab workers Health workers Counselors
)
HIV Testing Occurs in a Variety of ettings
STI Clinics
TB Clinics
Hospitals
T&C
ANC
Blood
Banks
Surveillance
!rovide care
to HIV"affected
#ersons
!rovide care
to HIV"affected
#ersons
Provide AR
treat!ent to
HI"in#ected
persons
Provide AR
treat!ent to
HI"in#ected
persons
Prevent HI
In#ections
Prevent HI
In#ections
Lab workers Health workers Counselors
*
$%#ansion of Testing ervices

Integrate HIV
laboratory services
and diagnostics fully
into national health
laboratory structures

+acilitate testing in
non#traditional
settings

&onsider all our

testing options
Lab workers Health workers Counselors
,
Initiate treat!ent with ARs
!ini!al dia$nostics re%uired
&onitor e##ectiveness o# ARs
with dia$nostics 'viral load(
C)*+ and sa#et, with basic
laborator, tests
-se HI rapid dia$nostic test to identi#,
an HI in#ection
&se of HIV Testing Technologies in the
&se of HIV Testing Technologies in the
Continuu' of Care
Continuu' of Care
Lab workers Health workers Counselors
-
#ectru' of HIV Tests

HIV diagnosis .Antibody/Antigen testing0

!n"yme Immunoassays .!IAs0

(apid tests

1estern blot .120

!arly diagnosis in infants

p2)

Initiation and monitoring of A('

&D)

Viral 3oad
Lab workers Health workers Counselors
4
Challenges of HIV Testing

!arly detection of seroconversion

!arly detection in infants born to HIV positive

mothers

!ffect of HIV subtypes on test performance

Impact of other health conditions on test

performance

5roduct specific e6uipment

'echnical s7ill
Lab workers Health workers Counselors
8
$n(y'e I''unoassays )$I*s+

9uantitative assay to measure HIV antibodies

:ost detect antibodies to HIV#; and HIV#2

Antigens coated in microwells

HIV Antigen / Antibody reaction is detected by color

change

Intensity of color reflects amount of antibody present

serum

%ome assays can detect both HIV antibody and

HIV antigen .close window period0

Issues:

%7illed lab technician

3arge volume testing

5roperly maintained e6uipment re6uired

Lab workers Health workers
;<
$n(y'e I''uno*ssays )$I*s+ , Cont-d
After several incubation and
wash steps, a color reaction
occurs if HIV antibody is present
An automated reader gives a
measurement of optical
density (presence of color)
for each well
Lab workers Health workers
;;
HIV .a#id Tests

9ualitative assay to detect HIV antibodies

:ost detect HIV ; and HIV 2

As reliable as !IAs

Issues:

%mall volumes

Validation of use

Appropriate training
Lab workers Health workers
;2
/estern 0lot 1 Line I''unoassays

=sed as supplemental test for

confirmation .only difficult cases0

Detects antibodies to specific

HIV antigens on cellulose strip

Issues:

:ultiple standards for

performance and
interpretation

!pensive

3imited commercial
availability
Lab workers Health workers
;$
HIV #23 *ntigen

&ore protein of the virus

!IA detects p2) antigen before antibody can be

detected

Detected 2 to $ wee7s after HIV infection

Detected about , days before antibody tests become reactive

=sed for:

Diagnosis of pediatric HIV#; infections

2lood ban7 safety .high incidence countries0

Issues:

3evel ) compleity

5roperly maintained e6uipment re6uired

Lab workers Health workers
;)
C43 T"Ly'#hocyte

&D) '#lymphocyte counts

used for:

Determining clinical
prognosis

Assessing criteria for

antiretroviral therapy

:onitoring therapy

:anual and automated

methods

Issues:

(e6uires high level of

technical s7ill for test
performance and
interpretation

5roperly maintained
e6uipment
Lab workers Health workers
;*
Viral Load

9uantitative molecular assay measures amount

of HIV in blood products

=sed to:

5redict disease progression

Assist with deciding when to initiate anti#retroviral

therapy

:onitors response to anti#retrovirals

Issues:

!pensive

3abor#intensive

%pecial facilities
Lab workers Health workers
;,





Co'#le%ity of HIV Tests Varies5
Co'#le%ity of HIV Tests Varies5

3evel ;: >o additional e6uipment and little or no

laboratory eperience needed

3evel 2: (eagent preparation or a multi#step

process is re6uired? &entrifugation or optimal
e6uipment

3evel $: %pecific s7ills such as diluting are re6uired

3evel ): !6uipment and trained laboratory technician

are re6uired
@1HA (eports @1HA (eports
Lab workers Health workers
;-
HIV .a#id Tests
#rovides
e%cellent tool
for e%#ansion
of services
Lab workers Health workers Counselors
;4
HIV .a#id Tests: *dvantages

Increases access to prevention .V&'0 and

interventions .5:'&'0

%upports increased number of testing sites

%ame#day diagnosis and counseling

(obust and easy to use

'est time under $< minutes

:ost re6uire no refrigeration

>one or one reagent

:inimal or no e6uipment re6uired

:inimum technical s7ill

Lab workers Health workers Counselors
;8
HIV .a#id Tests: 4isadvantages

%mall numbers for each test run

9uality Assurance/9uality &ontrol at

multiple sites

'est performance varies by product

(efrigeration re6uired by some products,

eBgB, &apillus

(eader variability in interpretation of

results

3imited end#point stability of test results

Lab workers Health workers Counselors
2<
0ody 6luids &sed for HIV .a#id Testing

%erum

5lasma

1hole blood

Aral fluids

Lab workers Health workers Counselors
2;
Three 6or'ats of HIV .a#id Tests

Immunoconcentration .flow#through
device0

Immunochromatography .lateral flow0

5article agglutination
Lab workers Health workers
22
How I''unoconcentration /or7s
HIV antibody links to bound HIV
peptide antigens forming the color
spot
HIV-
peptide
HIV-! peptide
Internal Control
Lab workers Health workers
2$
Tests 0ased on
I''unoconcentration
+low#'hrough Devices:

:ulti#%pot

Cenie II
Side view
Top view
Lab workers Health workers
2)
Non"reactive Reactive
.eading .esults: 8enie II
Lab workers Health workers Counselors
2*
Add Add
Sample Sample
Conjugate
C Control
line
T Test
Line
IgG Antibodies
HIV antibodies
Colloidal gold
conjugated to
HIV antigen
Anti-IgG/gold
antibodies
HIV antigen
How I''unochro'atogra#hy
/or7s
Lab workers Health workers
2,
Tests 0ased on
I''unochro'atogra#hy
Lateral .low )evices

Determine

Hema#%trip

Ara9uic7

=nigold
Sa!ple pad
HI Anti$en
Control
Speci!en .low
Lab workers Health workers
2-
Non"
Reactive
Reactive
a'#le !ad
.eading .esults: 4eter'ine
Control line Test line
Lab workers Health workers Counselors
24
.eading .esults: Ora9uic7
Non"
Reactive
Reactive
Lab workers Health workers Counselors
28
Anti"HI antibodies bind to the anti$en"coated
late/ particles
B
B
How !article *gglutination /or7s
Anti$en
Antibod,
Lab workers Health workers
$<
Tests 0ased On *gglutination
Agglutination devices:

&apillus

%erodia
Lab workers Health workers
$;
"on-
reactive
#eak
$eactive
%trong
$eactive
.eading .esults: Ca#illus
Lab workers Health workers Counselors
$2
There *re Only Three !ossible
Outco'es for ingle HIV *ntibody Tests
(eactive or D5ositiveE

'est band

&ontrol band
>on#reactive or D>egativeE

&ontrol band only

Invalid

>o control band present

'est has failedB (epeat with new deviceB

Lab workers Health workers Counselors
$$
$%ercise: Inter#reting Individual
HIV .a#id Test .esults

Refer to Participant Manual

Work alone to determine individual test

results

3 Minutes
Lab workers Health workers Counselors
$)
:ey Messages

HIV rapid tests can be as reliable as !IA

All tests re6uire attention to training,

supervision, and monitoring at points of
serviceB

As testing is epanding and decentrali"ed,

training, supervision, and monitoring must
follow accordingly and become all the
more importantB
Lab workers Health workers Counselors
$*
u''ary

1here is HIV rapid testing li7ely to occur

during an era of epansion of servicesF

1hat is the intended use for: !IAs,

1estern 2lot, p2) Antigen, &D), Viral
3oad

1hat are rapid testsF

1hy use rapid testsF

How do you read a result G reactive, non#

reactive or invalidF
Lab workers Health workers Counselors