Journal of Animal Research
  • Year: 2023
  • Volume: 13
  • Issue: 5

Quantitative real-time PCR as an alternative to plaque assay titration for recombinant baculovirus expressing porcine parvovirus VP2 gene

  • Author:
  • Tripti Pande1, A.K. Tiwari2, Sudhir Singh1, Mudasir M. Rather1, Vasavi Koppu1, Vikramaditya Upmanyu1,*
  • Total Page Count: 6
  • Published Online: Jul 31, 2024
  • Page Number: 637 to 642

1Division of Biological Standardization, ICAR-IVRI, Bareilly, Uttar Pradesh, India

2ICAR-CARI, Bareilly, Uttar Pradesh, India

*Corresponding author: V Upmanyu; E-mail: vupmanyu17@rediffmail.com

Online Published on 31 July, 2024.

Abstract

Baculovirus expression system having post translational modification is used for large scale production of foreign proteins. Viral titre determination is crucial for efficient protein production. Even though plaque assay and end-point dilution method are conventionally used for titre determination, a less tedious and time-saving method is required for viral titre determination. Recombinant baculovirus expressing VP2 of porcine parvovirus was transfected in SF-21 insect cells. A quantitative real-time PCR was optimized for r-baculovirus titre determination and correlated with plaque assay method for its performance. The baculovirus DNA qPCR Ct values and corresponding PFU/mL showed strong correlation having value of r =99.71 at 95% confidence interval.

• qPCR is a simpler and time-saving method that can be used for baculovirus titration.

• Ct value based copy no. strongly correlates with viral titre based on plaque assay.

Keywords

Baculovirus, Real-time PCR, Plaque assay, Insect cell, Porcine parvovirus, VP2, SF-21 cell line