Dr. Kate TempletonSPAIIN –Stirling 2011

OverviewHIV Infection

Tests used in diagnostic labs

How to access them

Challenges

HIV prevention

Non –conformity to existing strategies

An

ti-HIV

antib

ody

An

ti-HIV

antib

ody

An

ti-HIV

antib

ody

An

ti-HIV

antib

ody

Anti-HIV Anti-HIV antibodyantibody

Anti-HIV Anti-HIV antibodyantibody

Sero

con

vers

ion

Sero

con

vers

ion

illn

ess

illn

ess

Sero

con

vers

ion

Sero

con

vers

ion

illn

ess

illn

ess

Asymptomatic phaseAsymptomatic phaseAsymptomatic phaseAsymptomatic phase

Symptomatic Symptomatic phasephase

Symptomatic Symptomatic phasephase

AIDSAIDSAIDSAIDS

DeathDeathDeathDeath

Imm

un

olog

ical

fit

nes

sIm

mu

nol

ogic

al f

itn

ess

Imm

un

olog

ical

fit

nes

sIm

mu

nol

ogic

al f

itn

ess

0000 1111 5-30+5-30+5-30+5-30+ +1+1+1+1Years Years (average)(average)Years Years (average)(average)

Historic Major Historic Major Diagnostic Diagnostic

OpportunitiesOpportunities

Historic Major Historic Major Diagnostic Diagnostic

OpportunitiesOpportunities

List of testsHIV screening test

4th generationEIA formatIncludes Antigen as well as AntibodyOn large automated machines

HIV confirmation Blot2nd EIA

HIV RNA – viral loadHIV proviral DNA – used to detect HIV in

children <18 months old

Slide 5

RNARNARNARNA

AgAgAgAg

IgMIgMIgMIgM

IgGIgGIgGIgG

‘Qu

anti

ty’

‘Qu

anti

ty’

viral viral spreadspreadviral viral

spreadspread

local viral local viral growthgrowth

local viral local viral growthgrowth

00 11 22 33 44 55 66 1212 2424Weeks post-infectionWeeks post-infection

avidityavidity

IgG3

DNADNADNADNA

55 550000 10101010 15151515 20202020daysdaysdaysdays

Clonesystems Detect-HIV v1

Clonesystems Detect-HIV v1

InnotestHIV-1/-2

Murex ICE

HIV 1+2

Murex ICE

HIV 1+2

Ear

lies

t H

IV d

etec

tion

Ear

lies

t H

IV d

etec

tion

Biotest HIV 1/2recombinant

Biotest HIV 1/2recombinant

Vironostika HIV Uniform II Ag/AbVironostika HIV

Uniform II Ag/Ab

Enzygnost HIV Integral

Enzygnost HIV Integral

VIDAS HIV DUO

VIDAS HIV DUO

GENSCREEN PLUS HIV Ag-

Ab

GENSCREEN PLUS HIV Ag-

Ab

Architect HIV Ag/Ab ComboArchitect HIV Ag/Ab Combo

Murex HIV Ag/Ab

combionation

Murex HIV Ag/Ab

combionation

OrthoOrthoAb-capt.Ab-capt.ELISAELISA

OrthoOrthoAb-capt.Ab-capt.ELISAELISA

Vitros ECiVitros ECianti-HIV anti-HIV

1/21/2

Vitros ECiVitros ECianti-HIV anti-HIV

1/21/2Murex HIV Murex HIV

1/21/2VK84/85VK84/85

Murex HIV Murex HIV 1/21/2

VK84/85VK84/85

IMx IMx HIV1/2 HIV1/2 III plusIII plus

IMx IMx HIV1/2 HIV1/2 III plusIII plus

MurexMurexHIV HIV 1.2.01.2.0

GE94/95GE94/95

MurexMurexHIV HIV 1.2.01.2.0

GE94/95GE94/95

AxSYM AxSYM HIV 1/2 HIV 1/2

gOgO

AxSYM AxSYM HIV 1/2 HIV 1/2

gOgO

AbbottAbbott3rd gen 3rd gen

PlusPlus

AbbottAbbott3rd gen 3rd gen

PlusPlus

PasteurPasteurGenscreenGenscreenVersion 2Version 2

PasteurPasteurGenscreenGenscreenVersion 2Version 2

Access HIVAccess HIV1/2 NEW1/2 NEW

Access HIVAccess HIV1/2 NEW1/2 NEW

VironostikaVironostikaHIV Uniform HIV Uniform

II plus OII plus O

VironostikaVironostikaHIV Uniform HIV Uniform

II plus OII plus O

Biotest Biotest Anti-HIV Anti-HIV TETRA TETRA ELISAELISA

Biotest Biotest Anti-HIV Anti-HIV TETRA TETRA ELISAELISA

EnzygnostEnzygnostHIV 1/2 HIV 1/2

plusplus

EnzygnostEnzygnostHIV 1/2 HIV 1/2

plusplusE

arli

est

anti

-HIV

det

ecti

onE

arli

est

anti

-HIV

det

ecti

on

HIV testingAntenatal booking

HIV – 4th genHBsAgSyphilis IgGRubella IgG

Any HIV screen 4th gen HIV assayCaution – maternal HIV antibody can be

present for 15-24 months

Assay 2- 2Assay 2- 2ndnd EIA EIAAssay 2- 2Assay 2- 2ndnd EIA EIA

A1+ A2 +A1+ A2 +A1+ A2 +A1+ A2 +A1+ A2-A1+ A2-

Repeat A1 and A2Repeat A1 and A2A1+ A2-A1+ A2-

Repeat A1 and A2Repeat A1 and A2

A1+ A2+A1+ A2+A1+ A2+A1+ A2+ A1+ A2-A1+ A2-A1+ A2-A1+ A2-

Assay 3- blotAssay 3- blotAssay 3- blotAssay 3- blot

A1+ A2+ A3+A1+ A2+ A3+A1+ A2+ A3+A1+ A2+ A3+

report positivereport positivereport positivereport positive consider consider indeterminateindeterminate

consider consider indeterminateindeterminate

A1+ A2+ A3-A1+ A2+ A3-A1+ A2+ A3-A1+ A2+ A3- A1+ A2- A3+A1+ A2- A3+A1+ A2- A3+A1+ A2- A3+ A1+ A2- A3-A1+ A2- A3-A1+ A2- A3-A1+ A2- A3-

High riskHigh riskHigh riskHigh risk Low risk:Low risk:negativenegative

Low risk:Low risk:negativenegative

• Low prevalenceLow prevalence• asymptomaticasymptomatic• Low prevalenceLow prevalence• asymptomaticasymptomatic

A1 +A1 +A1 +A1 +

Screening EIAScreening EIAScreening EIAScreening EIA

A1-A1-report negativereport negative

A1-A1-report negativereport negative

A1- A2-A1- A2-report negativereport negative

A1- A2-A1- A2-report negativereport negative

Parry, bioMérieux Symposium, Oct 2006 Slide 10

Children born in UK & Ireland to HIV infected women,Children born in UK & Ireland to HIV infected women,reported by end of September 2005, likely infection status reported by end of September 2005, likely infection status

0

200

400

600

800

1000

1200

pre 1992 1993 1995 1997 1999 2001 2003 2005

year of birth

not infected indeterminate infected

Pat Tookey, Institute of Child Health

ZDV ZDV monotherapymonotherapy

ZDV ZDV monotherapymonotherapy

Antenatal Antenatal screening screening roll-outroll-out

Antenatal Antenatal screening screening roll-outroll-out

Slide 11

IgGIgGAnti-Anti-HIVHIV

reactireactivityvity

IgGIgGAnti-Anti-HIVHIV

reactireactivityvity

IgA/MIgA/MAnti-Anti-HIVHIV

IgA/MIgA/MAnti-Anti-HIVHIV

HIV RNAHIV RNAHIV RNAHIV RNA

P24 P24 AgAgP24 P24 AgAg

Rea

ctiv

ity‘

/ Qu

anti

ty’

Rea

ctiv

ity‘

/ Qu

anti

ty’

Rea

ctiv

ity‘

/ Qu

anti

ty’

Rea

ctiv

ity‘

/ Qu

anti

ty’

IgA/MIgA/MAnti-Anti-HIVHIV

IgA/MIgA/MAnti-Anti-HIVHIV

IgGIgGAnti-Anti-HIV HIV titretitre

IgGIgGAnti-Anti-HIV HIV titretitre

0000 1111 2222 3333 4444 5555 6666 …….12.12…….12.12 ……..18..18……..18..18Months post-partumMonths post-partumMonths post-partumMonths post-partum

HIV proviral HIV proviral DNADNA

HIV proviral HIV proviral DNADNA

HIV testing to exclude HIV infectionBHIVA/CHIVA guidelinesEDTA blood- baby

At birth within 1 day6 weeks ( 2-weeks after stopping Rx)3 monthsNeed 2 HIV proviral DNA while off therapyAntibody

EDTA blood – MotherHIV genetically diverse – check assay integrity against

Mothers specific virus.HIV viral load is optional – can give false positives/ or

more sensitiveFollowing exposure in genital tract

Summary of testingTime HIV ProV HIV Ab Baby/

Mother

Samples

Required

0-48 hrs

(not cord

blood)

Mother EDTA

min 1 ml

Baby EDTA

min 1 ml

6

weeks**

Baby EDTA

min 1 ml

12

weeks

Baby EDTA

min 1 ml

15-18

months

Baby EDTA

min 1 ml

HIV proviral DNA testingEDTA samples should be sent need prior telephone

notification  Specialist Virology CentreDepartment of Laboratory Medicine Royal Infirmary of Edinburgh51, Little France CrescentEdinburghEH16 4SA BBV team can be contacted on 0131 242 7129Duty Virologist can be contacted on 0131 242

6086 or 0131 536 1000, bleep 5981

Laboratory processingEDTA blood should be <24 hours old by time

in Edinburgh lab – preferably sooner.Keep at room temperatureCan arrive:

Monday –Friday 0900 -2000Saturday – 0900-1200

Current turnaround time is 7-10 daysAny positive results are phoned to referring

clinician

Final pointHIV protocol for access to HIV testing to

exclude infection in babies should be available via SPAIIN website.