Development of a set of the "Amlisense-HCV-genotype" reagents designed for determining the hepatitis С viruses of subtypes 1a, 1b, 2a, and 3a


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Abstract

Multiple alignments of 119 nucleotide sequences of isolates of hepatitis С 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, 1 b, 2a, and 3a subtypes. A PCR kit of reagents was designed for the amplification of cDNA HCV with selected typespecific 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 plasm id 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.

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Copyright (c) 2003 Gushchin A.E., Noskova О.М., Shipulin G.A.

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