A simple PCR with different 3' ends of the third primer for detection of defined point mutations: HCV genotyping as an example.
In this article, we describe a useful modification of the polymerase chain reaction for amplification applicable to hepatitis C virus genotyping and determination of its subtypes. The method is fast, cheap and simple for detection of any known point mutation, and could be used in every laboratory with experience in polymerase chain reaction technique. We could differentiate hepatitis C virus subtype 1b from other subtypes and 2b from 2a and other subtypes as well. We could also differentiate hepatitis C type 3 using a type-specific oligonucleotide from 3a subtype, thus covering the most common hepatitis C virus (sub)types present in the European region.
Alavantic
D
D
VINCA Institute, Laboratory for Radiobiology and Molecular Genetics, Belgrade, Yugoslavia. adragan@rt270.vin.bg.ac.yu
Glisic
S
S
Radovanović
N
N
Romić
M
M
D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry*; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood
D017931:DNA Primers; D012367:RNA, Viral