A technique for capturing broad subtypes and circulating recombinant             forms of HIV-1 based on anionic polymer-coated magnetic beads.
Magnetic beads coated with an anionic polymer, poly(methyl vinyl ether-maleic             anhydrate) [poly(MVE-MA)], were used in a method to capture human immunodeficiency             virus type-1 (HIV-1). The beads were incubated with either HIV-1-infected cell             culture medium or plasma from HIV-1 infected individuals and separated from the             supernatant by applying a magnetic field. After thorough washing, adsorption of             HIV-1 by the beads was confirmed by reverse transcription (RT)-polymerase chain             reaction (PCR), real-time PCR, enzyme-linked immunosorbent assay and western blotting.             The results confirmed the presence of envelope, polymerase, Nef and the viral             genome of HIV-1. Furthermore, various subtypes and circulating recombinant forms             (CRFs) of HIV-1 including subtype B, C and CRF01_AE and the immature form of subtype             B HIV-1 could be captured. Preincubation with neutralizing antibody against HIV-1             envelope gp41 decreased the capture efficiently, suggesting that poly(MVE-MA)             binds HIV-1 via gp41. We believe that this capture procedure will be a valuable             tool for detecting various types of HIV-1 in both clinical and experimental samples.
Department of Virology, Center for Infectious Disease Control,                     Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871,                     Japan.
Akikazu
A
Sakudo
Kazuyoshi
K
Ikuta
D000838:Anions; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015658:HIV Infections / Q000175:diagnosis* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008280:Magnetics; D011108:Polymers; D016865:Reassortant Viruses / Q000235:genetics / Q000302:isolation & purification; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000838:Anions; D011108:Polymers