Title: A genotypic assay for the amplification and sequencing of integrase from diverse HIV-1 group M subtypes
Authors: Van Laethem, Kristel ×
Schrooten, Yoeri
Covens, Kris
Dekeersmaeker, Nathalie
De Munter, Paul
Van Wijngaerden, Eric
Van Ranst, Marc
Vandamme, Anne-Mieke #
Issue Date: Nov-2008
Publisher: Elsevier/North-Holland Biomedical Press
Series Title: Journal of Virological Methods vol:153 issue:2 pages:176-181
Conference: European HIV Drug Resistance Workshop: From Basic Science to Clinical Implications edition:6 location:Budapest date:26-28 March 2008
Abstract: Recently, the Food and Drug Administration (FDA) of the USA approved the first integrase inhibitor for inclusion in treatment regimens of HIV-1 patients failing their current regimens with multi-drug resistant strains. However, treatment failure has been observed during integrase inhibitor-containing therapy. Several mutational pathways have been described with signature mutations at integrase positions 66, 92, 148 and 155. Therefore, a genotypic assay for the amplification and sequencing of HIV-1 integrase was developed. The assay displayed a detection limit of 10 HIV-1 III(B) RNA copies/ml plasma. As the HIV-1 pandemic is characterised by a large genetic diversity, the new assay was evaluated on a panel of 74 genetically divergent samples belonging to the following genetic forms A, B, C, D, F, G, J, CRF01-AE, CRF02-AG, CRFF03-AB, CRF12-BF and CRF13-cpx. Their viral load ranged from 178 until >500,000 RNA copies/ml. The amplification and sequencing was successful for 70 samples (a success rate of 95%). The four failures were most probably due to low viral load or poor quality of RNA and not to subtype issues. Some of the sequences obtained from integrase inhibitor-naïve patients displayed polymorphisms at integrase positions associated with resistance: 74IV, 138D, 151I, 157Q and 163AE. The relevance of these polymorphisms in the absence of the signature mutations remains unclear.
ISSN: 0166-0934
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Clinical and Epidemiological Virology (Rega Institute)
Laboratory for Clinical Infectious and Inflammatory Disorders
Molecular and Vascular Biology
× corresponding author
# (joint) last author

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