[Development of "Amplisens-HCV-genotype" reagent set for identification of hepatitis C virus genotypes 1a, 1b, 2a and 3a].
Multiple alignments of 119 nucleotide sequences of isolates of hepatitis C virus (HCV) were carried out to choose the type-specific primers for the 5'-ultra-core fragment of viral genome for the purpose of detecting the HCV 1a, 1b, 2a, and 3a subtypes. A PCR kit of reagents was designed for the amplification of cDNA HCV with selected type-specific primers and for making the electrophoresis in agarous gel. The kit comprises the positive control samples, i.e. HCV genome fragments, subtypes 1a, 1b, 2a and 3a, cloned in the plasmid vector. 440 cDNAHCV samples were simultaneously tested by using the worked out reagents' set and according to the method of Ohno et al. The results were found to be concordant in 336 cases, and were discordant in 4 samples. A sequencing of the PCR products and phylogenetic analysis showed that 1 sample belonged to subtype 4a, 2 samples belonged to subtypes 2k and 1 sample--to subtype 31.
Gushchin
A E
AE
Hoskova
O M
OM
Shipulin
G A
GA
D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D016679:Genome, Viral; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity
D017931:DNA Primers; D018076:DNA, Complementary; D011933:Reagent Kits, Diagnostic