Abstract：
:A long RT-PCR (LRP) protocol was developed recently for robust amplification of a near full-length HCV genomic sequence from clinical samples, followed by efficient cloning [Fan, X., Xu, Y., Di Biceglie, A.M., 2006. Efficient amplification and cloning of near full-length hepatitis C virus genome from clinical samples. Biochem. Biophys. Res. Commun. 346, 1163-1172]. In the present study, the LRP protocol has been estimated for its error rate and the validation by sequencing fully the near full-length HCV inserts from six recombinant clones derived from a patient sample with complex viral diversity. These sequences were compared with the near full-length HCV sequence that was generated by direct sequencing of multiple overlapped PCR products from the same sample, referred to as the population sequence. Comparative analysis confirmed the artificial nature of the PCR-assembled population sequence and identified potential domains for linked viral mutations. The data also suggested that the hypervariable region 1 (HVR1) may be a biological marker for the phenotype at the quasispecies level. These observations emphasize the significance of the use of near full-length genomic sequences for HCV genetic studies and for reverse genetic analysis using authentic quasispecies variants.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Zhou D,Fan X,Tan D,Xu Y,Tavis JE,Di Bisceglie AMdoi
10.1016/j.jviromet.2006.12.002subject
Has Abstractpub_date
2007-05-01 00:00:00pages
220-4issue
2eissn
0166-0934issn
1879-0984pii
S0166-0934(06)00446-0journal_volume
141pub_type
杂志文章abstract：:In vitro enzymatic amplification was applied to detect hepatitis B virus (HBV) DNA sequences in serum. This technique, known as the polymerase chain reaction (PCR) was used to amplify a 128 bp DNA fragment including a 112 nucleotide long sequence complementary to a region in the S gene of the HBV genome. Amplified sam...
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journal_title：Journal of virological methods
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pub_type： 临床试验,杂志文章
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
pub_type： 杂志文章
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journal_title：Journal of virological methods
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