Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan.
Int J Mol Med. 2012 Aug;30(2):437-42. doi: 10.3892/ijmm.2012.1009. Epub 2012 May 22.
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.
用带负电荷的聚合物聚（甲基乙烯基醚-马来酸酐）[poly（MVE-MA）]包被的磁珠，用于捕获人类免疫缺陷病毒 1 型（HIV-1）。将珠粒与 HIV-1 感染的细胞培养物或来自 HIV-1 感染者的血浆孵育，然后通过施加磁场将其与上清液分离。彻底洗涤后，通过逆转录（RT）-聚合酶链反应（PCR）、实时 PCR、酶联免疫吸附测定和 Western blot 确认珠粒对 HIV-1 的吸附。结果证实了 HIV-1 的包膜、聚合酶、Nef 和病毒基因组的存在。此外，还可以捕获各种 HIV-1 亚型和循环重组形式（CRF），包括亚型 B、C 和 CRF01_AE 以及亚型 B HIV-1 的不成熟形式。用 HIV-1 包膜 gp41 的中和抗体预先孵育可有效降低捕获效率，表明 poly（MVE-MA）通过 gp41 结合 HIV-1。我们相信，这种捕获程序将成为在临床和实验样本中检测各种类型 HIV-1 的有价值的工具。
