title: Application of real-time quantitative polymerase chain reaction to monitoring infection of classic swine fever virus and determining optimal harvest time in large-scale production.
abstract: Due to the non-cytopathogenic replication of classical swine fever virus (CSFV) in cell culture, large-scale production of CSFV using bioreactor system remains the problem of monitoring the time of maximum virus production for optimal harvest. In this study, we proposed the application of real-time quantitative PCR assay to monitoring the progress of CSFV infection and yield determination in large scale. The region of NS5B of CSFV responsible for CSFV genome replication was used for the designation of primers and probe. Viral titers determined by the real-time quantitative PCR assay were compared with the conventional cell-culture based method of immunofluorescent staining. Results from large scale production show that a similar profile of CSFV production was successfully outlined by real-time quantitative PCR and virus yields were comparable to the results from immunofluorescent staining assay. By using this method, an optimal harvesting time of the production could be rapidly and precisely determined leading to an improvement in virus harvest.
authors.affiliation: Department of Biotechnology, Fooyin University, 151 Jinxue Road, Daliao District, Kaohsiung City 83102, Taiwan, ROC. Electronic address: dr005@fy.edu.tw.
authors.forename: Ya-Ching
authors.initials: YC
authors.lastname: Lin
authors.forename: Sheng-Chi
authors.initials: SC
authors.lastname: Wu
authors.forename: Ming-Yu
authors.initials: MY
authors.lastname: Yang
authors.forename: Guan-Ting
authors.initials: GT
authors.lastname: Chen
authors.forename: Tzung-Han
authors.initials: TH
authors.lastname: Li
authors.forename: Ming-Yi
authors.initials: MY
authors.lastname: Liau
mesh_terms: D000818:Animals; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000254:growth & development* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D015345:Oligonucleotide Probes / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013552:Swine; D019562:Viral Load / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics; D014776:Virus Cultivation / Q000379:methods
chemical_list: D017931:DNA Primers; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D014764:Viral Proteins
keywords: CSFV; Mass production; Non-cytopathogenic; Optimum harvest time; Real-time quantitative PCR assay; Vaccine