Mesh : Biotinylation Blood / virology Blood Chemical Analysis / methods HIV / genetics isolation & purification Hepacivirus / genetics isolation & purification Humans Molecular Diagnostic Techniques Nucleic Acid Hybridization Oligonucleotides / metabolism Polymerase Chain Reaction RNA, Viral / blood genetics isolation & purification metabolism

来  源:   DOI:10.1007/978-1-61779-937-2_3

Abstract:
The quantification of Hepatitis C Virus (HCV) and Human Immunodeficiency Virus (HIV) in whole blood provides several advantages over the quantification in plasma samples. The application of small samples of capillary blood allows for application in point-of-care diagnostic testing methods.Here we describe two protocols of extracting viral RNA from small samples of whole blood by hybridization to biotinylated LNA-modified 2\'-O-Methyl-RNA or to biotinylated DNA, indirect capturing to streptavidin-coated beads, and subsequent quantification by one-step non-nested qRT-PCR. Further, we provide some general guidelines on extraction and quantification of HIV and HCV in small volume whole blood samples.
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