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. 2010 Feb 6;12(1):62–69. doi: 10.1007/s11894-009-0088-1

Table 2.

Genotypic and phenotypic assays

  Advantages Disadvantages
Genotypic assays
 Direct PCR sequencing Thorough identification of all mutations Poor sensitivity (∼ 20%)
 Restriction fragment-length polymorphism Sensitive to ∼ 5% Requires separate endonuclease reactions for each mutation
 Reverse hybridization line probe assay Sensitive Requires specific probe for every mutation
Commercially available
 DNA microchip Able to detect new mutations Expensive
Not widely available
 Mass spectrometric analysis Very sensitive Requires new primer for each mutation
 Ultra-deep pyrosequencing Sensitive to 0.1% Early in development
 
Phenotypic assays
 Transient transfection with site-specific mutagenesis Rapid Not applicable for multiple or complex mutations
Good for single mutations
 Whole HBV genome amplification/cloning and transfection Whole HBV genome variability Requires cloning step
Cross-resistance testing
 Cell transduction using Baculovirus vector High levels of replication Multiple vectors required
Cross-resistance testing
 Permanent cell lines with expression of HBV mutant genome Reproducible New cell line needed for new mutations
Cross-resistance testing

HBV—hepatitis B virus; PCR—polymerase chain reaction