Probes and methods for hepatitis C virus typing using single probe analysis

A single-tube method for determining the type of a hepatitis C virus (HCV) and an HCV viral load in a sample, the method comprising:

• a) amplifying a portion of the HCV genome, from the sample via asymmetric PCR, thereby producing a limiting amplicon strand and an excess amplicon strand;
• b) hybridizing said limiting amplicon strand, with an amplicon quantitation probe and quantifying the amount of said limiting amplicon strand;
• c) hybridizing said excess amplicon strand, with a typing probe to form a target hybridization complex, wherein:
• i) said typing probe is complementary or partially complementary to a nucleotide sequence within an HCV genome;
• ii) said nucleotide sequence within an HCV genome shows sequence heterogeneity among at least five HCV genotypes or at least six HCV subtypes;
• and,
• iii) hybridization complexes comprising said typing probe and the at least five HCV genotypes or the at least six HCV subtypes have a distinguishing hybridization property that differentiates each of the HGV genotypes or HCV subtypes;
• d) measuring said distinguishing hybridization property of the target hybridization complex, and,
• e) correlating said measured distinguishing hybridization property of the target hybridization complex with a similar property of an HCV genotype or subtype wherein all the steps above are performed in a single tube.