How do we measure HIV drug resistance. Monique Nijhuis
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1 How do we measure HIV drug resistance Monique Nijhuis
2 Measure HIV drug resistance Genotypic resistance test vs Phenotypic resistance test
3 Measure HIV drug resistance Genotypic resistance test vs Phenotypic resistance test Examen if HIV has changes (mutations) in the genetic structure
4 Measure HIV drug resistance Genotypic resistance test vs Phenotypic resistance test Recombinant phenotypic test (part of the virus with mutations) Examen if HIV has changes (mutations) in the genetic structure Examen if drugs can inhibit the replication of HIV
5 Measure HIV drug resistance: blood sample Centrifugation of blood sample
6 Measure HIV drug resistance: blood sample HIV genotypic resistance test recombinant phenotypic test Centrifugation of blood sample
7 Measure HIV drug resistance: blood sample HIV genotypic resistance test recombinant phenotypic test genotypic resistance test phenotypic resistance test HIV-infected cells (alive) Centrifugation of blood sample
8 Measure HIV drug resistance: blood sample HIV genotypic resistance test recombinant phenotypic test genotypic resistance test phenotypic resistance test HIV-infected cells (alive) Centrifugation of blood sample Dried blood spot HIV HIV-infected cells (dead) genotypic resistance test recombinant phenotypic test
9 Plasma Genotypic drug resistance test
10 Genotypic drug resistance test Plasma DBS cells Proviral DNA
11 Genotypic drug resistance test Plasma DBS cells Proviral DNA
12 Genotypic drug resistance test pro/cons Pro: -relatively fast and cheap -relatively successful on low level viremia samples
13 Genotypic drug resistance test pro/cons Pro: -relatively fast and cheap -relatively successful on low level viremia samples Hofstra, personal communication
14 Genotypic drug resistance test pro/cons Pro: -relatively fast and cheap -relatively successful on low level viremia samples Con: -indirect measurement of drug resistance -need to have a database with known impact of mutations
15 Interpretation of genotypic resistance
16 Interpretation of genotypic resistance Interpret impact of (combinations) of resistance mutations: Algorithms
17 Genotypic drug resistance test pro/cons Pro: -relatively fast and cheap -relatively successful on low level viremia samples Con: -indirect measurement of drug resistance -need to have a database with known impact of mutations -resistance may depend on the interaction of mutations -virus background, subtype -challenging for new drugs and especially new drug classes -Phenotypic drug resistance assay (recombinant phenotypic test)
18 Recombinant phenotypic resistance test
19 Suppression Recombinant phenotypic resistance test 100% 75% 50% Fold resistance IC 50 IC 50 Patient isolate Wildtype 25% 0% Drug concentration (log)
20 Genotypic drug resistance test pro/cons Pro: -relatively fast and cheap -relatively successful on low level viremia samples Con: -indirect measurement of drug resistance -need to have a database with known impact of mutations -resistance may depend on the interaction of mutations -virus background, subtype -challenging for new drugs and especially new drug classes -Phenotypic drug resistance assay (recombinant phenotypic test) -Phenotypic drug resistance assay
21 Measure HIV drug resistance: blood sample HIV genotypic resistance test genotypic resistance test phenotypic resistance test HIV-infected cells (alive) Centrifugation of blood sample genotypic resistance test Dried blood spot HIV HIV-infected cells (dead)
22 Phenotypic drug resistance test Virus culture HIV-infected cells (alive)
23 Phenotypic drug resistance test Pro: -direct measurement of drug resistance -you do not need to know the location of the mutation Con: -relatively time consuming and expensive -clinically relevant levels of drug resistance are not always available -require specialized laboratory (BSLIII)
24 Biosafety level III laboratory double door entry, inward air flow, biosafety cabinets, on site autoclave
25 Choose your resistance test wisely Genotypic resistance test: -known antiretroviral drugs with good interpretation algorithms Phenotypic resistance test: -virological failure can not be explained based on genotypic resistance profile -new drugs or new drug classes
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