Guidelines for the quantification of HIV and HCV in small volume whole blood samples.
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.
Division of Gastroenterology, Hepatology, and Infectious Diseases, Department of Internal Medicine II, Jena University Hospital, Jena, Germany. T.Bruns@bham.ac.uk
Tony
T
Bruns
Katrin
K
Steinmetzer
D019921:Biotinylation; D001769:Blood / Q000821:virology*; D001774:Blood Chemical Analysis / Q000379:methods*; D006678:HIV / Q000235:genetics / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D009841:Oligonucleotides; D012367:RNA, Viral; C477371:locked nucleic acid