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Journal of Clinical Microbiology, April 2004, p. 1428-1433, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1428-1433.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development, Evaluation, and Validation of an Oligonucleotide Probe Hybridization Assay To Subtype Human Immunodeficiency Virus Type 1 Circulating Recombinant Form CRF02_AG

Harr F. Njai,1 Gert Van der Auwera,2 Chiambah A. Ngong,1 Leo Heyndrickx,1 Souleymane Sawadago,3 Hilton Whittle,4 Phillipe Nyambi,5 Robert Colebunders,1 Guido van der Groen,1 and Wouter Janssens1,6*

Department of Microbiology, Institute of Tropical Medicine, Antwerp,1 Department of Plant Systems Biology, University of Ghent, Ghent,2 Flanders Interuniversity Institute for Biotechnology (VIB), Zwijnaarde, Belgium,6 Projet RETRO-CI, Abidjan, Côte d'Ivoire,3 Medical Research Council Laboratories, Fajara, The Gambia,4 Department of Pathology, New York University School of Medicine, New York, New York5

Received 4 June 2003/ Returned for modification 25 September 2003/ Accepted 24 December 2003

We have developed and validated an oligonucleotide probe hybridization assay for human immunodeficiency virus type 1 (HIV-1) circulating recombinant form (CRF) CRF02_AG. In the p17 coding region of the gag gene, a CRF02_AG-specific signature pattern was observed. Five working probes were designed to discriminate CRF02_AG infections from infections by all other documented subtypes and CRFs in an enzyme-linked immunosorbent assay-based oligonucleotide probe hybridization assay. Nucleic acids were extracted from a panel of HIV-1-positive plasma samples from Cameroon, Bénin, Côte d'Ivoire, Kenya, Zambia, and Belgium and from blood spots from The Gambia. CRF02_AG (n = 147) and non-CRF02 (n = 100) samples were analyzed to evaluate and validate the oligonucleotide probe hybridization assay. The CRF02_AG-specific oligonucleotide probe hybridization assay has a high sensitivity and specificity, with good positive and negative predictive values in regions of high and low prevalence. A validation of the assay with West and West Central African samples indicated a sensitivity of 98.4% and a specificity of 96.7%. The oligonucleotide probe hybridization assay as a diagnostic tool will allow for rapid screening for CRF02_AG. This could be used to track the HIV epidemic in terms of documenting the real prevalence of CRF02_AG strains and will complement efforts in vaccine development. Moreover, this technology can easily be applied in laboratories in developing countries.

* Corresponding author. Mailing address: Department of Microbiology, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium. Phone: 32 3 247 63 28. Fax: 32 3 247 63 33. E-mail: wjanssens{at}itg.be.

Journal of Clinical Microbiology, April 2004, p. 1428-1433, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1428-1433.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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