Method for the preparation of purified HCV RNA by exosome separation
AssigneeNovartis Vaccines and Diagnostics, Inc.
InventorsSergio Abrignani, Piero Pileri
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Abstract
The invention relates to a method for the isolation of hepatitis C virus. The method comprises the separation of particles termed exosomes from the blood plasma of an individual infected with hepatitis C virus (HCV) and the extraction or RNA from these exosome particles.
Claims21
A method for preparation of purified HCV RNA comprising separating exosome particles from the supernatant of a cell culture infected with HCV and extracting RNA from the exosome particles.
A method of diagnosing an individual as being infected with HCV, comprising obtaining a preparation of cells from the individual, preparing exosome particles from the cellular supernatant and testing the exosome particles for the presence of HCV RNA and proteins.
A method for preparing purified HCV RNA comprising:
- (a) providing plasma from a subject infected with HCV;
- (b) subjecting the plasma to differential centrifugation to produce an exosome-enriched supernatant;
- (c) incubating the supernatant with beads coated with an anti-CD81 or anti-CD82 antibody to isolate exosome particles;
- and
- (d) extracting HCV RNA from the exosome particles.
A method for preparing purified HCV RNA comprising:
- (a) providing plasma from a human infected with HCV;
- (b) subjecting the plasma to differential centrifugation to produce an exosome-enriched supernatant;
- (c) incubating the supernatant with magnetic beads coated with an anti-CD81 or anti-CD82 antibody to isolate exosome particles;
- and
- (d) extracting HCV RNA from the exosome particles using a viral extraction kit.
Dependencies are inferred from the claim text.