+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

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



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



Vaccine 31(47): 5565-5571



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.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 051612658

Download citation: RISBibTeXText

PMID: 24021314

DOI: 10.1016/j.vaccine.2013.08.075


Related references

Comparison of two real-time quantitative polymerase chain reaction strategies for minimal residual disease evaluation in lymphoproliferative disorders: correlation between immunoglobulin gene mutation load and real-time quantitative polymerase chain reaction performance. Hematological Oncology 32(3): 133-138, 2014

Application of Quantitative Real- Time Polymerase Chain Reaction for Noninvasive Genetic Monitoring. Journal of Wildlife Management 74(8): 1904-1910, 2010

Real-time quantitative polymerase chain reaction for virus infection diagnostics. Expert Opinion on Medical Diagnostics 2(10): 1155-1171, 2008

Detection of African swine fever, classical swine fever, and foot-and-mouth disease viruses in swine oral fluids by multiplex reverse transcription real-time polymerase chain reaction. Journal of Veterinary Diagnostic Investigation 27(2): 140-149, 2015

Development, optimization, and validation of a Classical swine fever virus real-time reverse transcription polymerase chain reaction assay. Journal of Veterinary Diagnostic Investigation 23(5): 994-998, 2011

Optimal sampling time after preparation of platelet concentrates for detection of bacterial contamination by quantitative real-time polymerase chain reaction. Vox Sanguinis 89(4): 208-214, 2005

A one-step real-time reverse transcription-polymerase chain reaction detection of classical swine fever virus using a minor groove binding probe. Veterinary Research Communications 34(4): 359-369, 2010

Diagnosis of feline leukaemia virus infection by semi-quantitative real-time polymerase chain reaction. Journal of Feline Medicine and Surgery 9(1): 8, 2007

Comparison of methods for improved RNA extraction from blood for early detection of Classical swine fever virus by real-time reverse transcription polymerase chain reaction. Journal of Veterinary Diagnostic Investigation 23(4): 727-735, 2011

Prospective monitoring of the Epstein-Barr virus DNA by a real-time quantitative polymerase chain reaction after allogenic stem cell transplantation. British Journal of Haematology 115(1): 105-111, 2001

Use of automated real-time reverse transcription-polymerase chain reaction (RT-PCR) to monitor experimental swine vesicular disease virus infection in pigs. Journal of Comparative Pathology 131(4): 308-317, 2004

Establishment and application of a TaqMan real-time quantitative reverse transcription-polymerase chain reaction assay for rubella virus RNA. Acta Biochimica et Biophysica Sinica 38(10): 731-736, 2006

Micro-droplet Digital Polymerase Chain Reaction and Real-Time Quantitative Polymerase Chain Reaction Technologies Provide Highly Sensitive and Accurate Detection of Zika Virus. Virologica Sinica 33(3): 270-277, 2018

Usefulness of quantitative real-time polymerase chain reaction in following up patients with Epstein-Barr virus infection after liver transplantation. Clinical Transplantation 14(4 Pt 1): 308-317, 2000

Establishment and application of nested real-time quantitative polymerase chain reaction assay for detection of hepatitis B virus covalently closed circular DNA. Zhonghua Shi Yan he Lin Chuang Bing du Xue Za Zhi 25(4): 307-309, 2011