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Protein Bead Array for the Detection of HIV-1 Antibodies from Fresh Plasma and Dried-Blood-Spot Specimens

¹ National HIV Immunology Laboratory, National HIV and Retrovirology Laboratories, Centre for Infectious Disease Prevention and Control, Health Canada, Ottawa, ON, Canada;² National Laboratory for HIV Reference Services, National HIV and Retrovirology Laboratories, Centre for Infectious Disease Prevention and Control, Health Canada, Ottawa, ON, Canada;³ National HIV and Retrovirology Laboratories, Centre for Infectious Disease Prevention and Control, Health Canada, Ottawa, ON, Canada

^aaddress correspondence to this author at: National HIV Immunology Laboratory, National HIV and Retrovirology Laboratories, Centre for Infectious Disease Prevention and Control, Bldg. 6, P.L. 0603B1, Tunney’s Pasture, Ottawa, Ontario, Canada K1A 0L2; fax 613-946-3237, e-mail sylvie_faucher@hc-sc.gc.ca

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Simultaneous multianalyte immunoassays offer advantages, including reduction of technical operations, execution time, and overall costs. The suspension array technology (SAT), which uses microfluorospheres with flow cytometry, is a multianalyte immunoassay that requires small specimen volumes, such as those obtained by less-invasive blood sampling approaches such as heel or finger sticks, as well as those obtained in pediatric specimens. The dried-blood-spot (DBS) technology has become an important screening tool for clinical and epidemiologic testing (1)(2)(3). It is particularly convenient in rural, resource-limited settings where trained personnel and adequate facilities for blood collection, processing, transport, and storage may not be available. This approach, however, provides limited specimen volume (5–6 µL of serum/6-mm punch) (2). Fortunately, SAT is well suited to support such a format (4)(5).

The range of applications for SAT includes antibody, oligonucleotide, peptide, and protein bead arrays (PBAs) (4)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). PBAs have been used for the simultaneous detection of serum antibodies to infectious agents for systemic candidiasis (18), herpes viruses (19), measles, mumps (20), and HIV (21)(22). The successful detection of HIV-1 serum antibodies by a four-protein bead array was first reported by Scillian et al. (21), who used beads of various sizes and proteins noncovalently coupled to beads. Reagents are now available that permit covalent coupling of proteins to beads and simultaneous analysis of up to 100 color-coded bead sets in a high-throughput, 96-well plate format. In addition, a new generation of flow cytometers compatible with SAT has recently been introduced, e.g., the Luminex-100 System (Luminex Corp.) and the . . . [Full Text of this Article]