Characterization of hepatitis C virus quasispecies by matrix-assisted laser desorption ionization-time of flight (mass spectrometry) mutation detection.
Hepatitis C virus (HCV), the causative agent of hepatitis C, frequently causes chronic infection. The mechanisms of viral persistence continue to be the object of investigation. An important aspect of HCV chronic infection is the quasispecies nature of the viral population, which has been particularly well documented in the hypervariable region 1 of the E2 glycoprotein. Recent studies show that characterization of the quasispecies diversity at the amino acid level can help to predict the outcome of HCV infection. Currently the accurate characterization of HCV quasispecies requires the cloning of PCR products, followed by the sequencing of many clones. In this study we present a new method to characterize HCV quasispecies, based on in vitro translation of the amplicons, followed by mass spectrometry analysis of the resulting peptide mix. The assay was used on reference HCV samples and on clinical samples. In principle, this method could be applied to other chronic viral infections in which quasispecies play a role.
Ayers
Melissa
M
Divisions of Microbiology. Gastroenterology and Nutrition, Hospital for Sick Children, Toronto, Ontario, Canada.
Siu
Karen
K
Roberts
Eve
E
Garvin
Alex M
AM
Tellier
Raymond
R
D002675:Child, Preschool; D005260:Female; D005838:Genotype; D016174:Hepacivirus / Q000145:classification* / Q000235:genetics*; D019698:Hepatitis C, Chronic / Q000821:virology*; D006801:Humans; D007223:Infant; D008297:Male; D009154:Mutation; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D014176:Protein Biosynthesis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
C400562:HVR1 protein, Hepatitis C virus; D010455:Peptides; D014764:Viral Proteins