HIV-1 and HIV-2 diagnostics AREVIR, Kln 11.04.2013 How different - PowerPoint PPT Presentation

HIV-1 and HIV-2 diagnostics AREVIR, Köln 11.04.2013

How different are they? 1, 2, 3…

Phylogenetic view of immune deficiency viruses

Nomenclature of HIV: types, groups and subtypes type HIV-1 group M- 1983 P -2009 N -1998 O -1994 subtype A B/D C F G H J K I II III IV V VI type HIV-2, 1985 A B C D E F G H group A1,A2 HIV unknown stolen from Lutz Gürtler

Forests and distribution of sooty mangabey monkeys (Cercocebus atys) and HIV-2 in West-Africa Forests and sooty mangabey HIV-2 Modified from Lutz Gürtler

Prevalence of HIV-2 groups in West-Africa: A and B in all countries, India only HIV-2:A, USA 180 cases Cap Verde Island

Epidemiology of HIV-2  Mainly in West-Africa  But also in countries associated to West- Africa  in Portugal, HIV-2 causes 4.5% of AIDS cases  in France, 1.8% of new HIV diagnoses between 2003 and 2006 by HIV-2  …

HIV diagnosis - methods  4th generation screening ELISA detect  antibodies against HIV-1/2  HIV-1 p24 antigen  Confirmation by immunoblot  Usually specific for HIV-1 or HIV-2  Not covering p24 component of screening  Validation of a 2-rapid-test system ?

HIV-2 diagnostic – different-2 ?

Western blot – specific?  There is substantial antibody cross-reactivity  There are HIV-1 immunoblots including a specific HIV-2 band • BUT is there co-infection of HIV-1/2 ?

How to detect HIV-2 definitely ?  Reactive HIV screening test needs confirmation by  Immunoblot  A second independant sample  Nucleid acid testing for HIV-1 • First viral load  Nucleid acid testing for HIV-2 ? • Independent proof by viral component

HIV Genotypischer Resistenzalgorithmus Deutschland HIV2EU collaboration Resistance algorithm

HIV2EU publication Charpentier, Charlotte, Ricardo Camacho, Jean Ruelle, Rolf Kaiser, Josef Eberle, Lutz Gürtler, Alejandro Pironti, Martin Stürmer, Françoise Brun-Vézinet, und Diane Descamps. „HIV-2EU-Supporting Standardized HIV-2 Drug Resistance Interpretation in Europe“. Clinical Infectious Diseases (2013). http://cid.oxfordjournals.org/content/early/2013/02/19/cid.cit104.short. M. Obermeier 5/2012

HIV-2 sequences from Los Alamos National Laboratories in the HIV-GRADE tool M. Obermeier 5/2012

Basis of the HIV-GRADE internet-tool  HIV-Alg module from Stanford-HIVdb  PERL-source-code is freely available  Software is in development since 1999  Algorithm Specification Interface (ASI) M. Obermeier 5/2012

Algorithm Specification Interface (ASI)  Rule-based algorithms can be described using xml-syntax  Xml-Files available for Stanford- HIVdb, ANRS and REGA HIV-GRADE can be described in a compatible format. M. Obermeier 5/2012

M. Obermeier 5/2012

M. Obermeier 5/2012

Workflow HIV2EU Sequence data Mutation list Alignment on Pan- Genotype Consensus Identification of available genes Identification of genotype and re-alignment on group specific consensus-sequence Identification of mutations Analysis of mutations based on the algorithm Detailled output Tabular output M. Obermeier 5/2012

HIV-2 tropism mutations Visseaux, B., M. Hurtado-Nedelec, C. Charpentier, G. Collin, A. Storto, S. Matheron, L. Larrouy, u. a. „Molecular Determinants of HIV-2 R5-X4 Tropism in the V3 Loop: Development of a New Genotypic Tool“. Journal of Infectious Diseases 205, Nr. 1 (2. Dezember 2011): 111–120. doi:10.1093/infdis/jir698. M. Obermeier 5/2012

German HIV-2 testing and treatment guidelines  HIV-GRADE and HIV2EU team were asked by the DAIG  Diagnostic guidance  Treatment guidance M. Obermeier 5/2012

M. Obermeier 5/2012

HIV-GRADE association  Thomas Berg, Medizinisches Labor Dr. Berg, Berlin  Patrick Braun, PZB, Aachen  Martin Däumer, Institut für Virologie, Köln  Josef Eberle, Pettenkofer-Institut, München  Robert Ehret, PZB Aachen  Rolf Kaiser, Institut für Virologie, Köln  Klaus Korn, NRZ für Retroviren, Erlangen  Claudia Kücherer, Robert Koch Institut, Berlin  Harm Müller,Labor Fenner, Hamburg  Christian Noah, Labor Lademannbogen, Hamburg  Martin Stürmer, Institut für Medizinische Virologie, Frankfurt  Alexander Thielen, Max Planck Institut Saarbrücken  Hauke Walter, NRZ für Retroviren, Erlangen

HIV2EU group  Charlotte Charpentier  Ricardo Camacho  Jean Ruelle  Rolf Kaiser  Josef Eberle  Lutz Gürtler  Alejandro Pironti  Martin Stürmer  Françoise Brun-Vézinet  Diane Descamps  Martin Obermeier M. Obermeier 5/2012