Journal of General Virology · 1991 · 79 citations · 24 references
Life SciencesInfectious DiseasesMicrobiologyMedical MicrobiologyMolecular VirologyVirologyViral ClearanceViral DiagnosticsViral HepatitisViral PathologyVirus PurificationVirus RemovalHepatitis Virus TestingParticle Size ReductionPore Size ControlViral FiltrationVirus Purity AssessmentVirus Separation
To estimate the particle size of hepatitis C virus (HCV), a major causative agent of post-transfusion non-A, non-B hepatitis, we filtered plasma or serum samples through microporous cellulose fibres with different pore sizes. The amount of HCV particles in samples before and after filtration was determined by a quantitative reverse transcriptase polymerase chain reaction (PCR) method. Since there is no quantitative biological assay for HCV, except for that in chimpanzees, the HCV titre obtained from the PCR method was used in an equation constructed previously for application to filtration experiments with a flavivirus which is distantly related to HCV. The particle was estimated to be between 30 and 38 nm in diameter, although the possibility remained that larger HCV particles or HCV aggregates with a diameter of more than 39 nm might exist. Double-step filtration through microporous cellulose fibres with a pore size of 35 nm reduced the HCV content to below levels detectable by our PCR method, indicating that it is possible to eliminate HCV particles by simple filtration techniques.
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Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
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