Dynamic changes of virus load in supernatant of primary CEK cell culture infected with different generations of avian infectious bronchitis virus strains Sczy3 as revealed by real-time reverse transcription-polymerase chain reaction

Abstract

ABSTRACT. Infectious bronchitis virus (IBV) can multiply effectively in chick embryo kidney (CEK) cells after adapting to the chick embryo. To investigate the dynamic changes in IBV load in the supernatant of primary CEK cells, we developed an SYBR Green I-based real-time polymerase chain reaction assay to quantify nucleic copy numbers of the IBV-Sczy3 strain. The 20, 54, and 87th generations of CEK-adapted IBV-Sczy3 strains were used to infect CEK cells, and then nucleic copy numbers in the samples of supernatant collected at 12, 24, 36, 48, 60, and 72 h were detected. The results showed that the rapid growth period of the virus load of all the 3 generations was approximately 12-36 h post- (2015) infection; the peak of the virus load appeared at 36 h post-infection and then decreased gradually in the order of 20th > 54th > 87th for the 3 generations of CEK-adapted strains; the dynamic change curve of the IBV load in the supernatant of primary CEK cells showed a single peak. The results of this study provide a useful reference for CEK-adapted IBV field strains and the production of CEK-attenuated IBV vaccine.

Changes of Antioxidant Function and the mRNA Expression Levels of Xanthine Oxidase in Primary Chick Kidney Cell Culture Caused by Nephropathogenic Infectious Bronchitis Virus Infection

“…Previous study documented that epithelial cell was the primary site of replication of avian NIBV, the increased level of virion in CK cells lead to destructions of the cellular structure and composition, which caused the cell membranes damage (Sun et al, 2014). Interestingly, the characteristic syncytia and plaque was observed in CEK cells after transfected by IBV Sczy3 virus (Wei et al, 2015) and intracytoplasmic brownish colouration was observed by immunoperoxidase in primary chick embryo chorioallontoic membrane cells after infection by IBV (Ghetas et al, 2015). The results are similar to our study: post infected by NIBV, the cytopathic effect was found and the cell viability of CK cells was significant reduced.…”

Antigenic and Pathogenic Characteristics of QX-Type Avian Infectious Bronchitis Virus Strains Isolated in Southwestern China

“…The amplicons were then inserted into pMD-19T cloning vector (Takara Bio-Inc, Dalian, China) and transformed into Escherichia coli TOP10 competent cells (Takara Bio-Inc, Dalian, China). The recombinant plasmid DNA was extracted, and its copy numbers were calculated according to a previous report [29].…”