Isolation of Infectious Bronchitis Virus in Primary cells of the Chick Embryo Chorioallantoic Membrane

Abstract

The susceptibility of the primary chick embryo chorioallontoic membrane cells to infectious bronchitis virus was evaluated after twenty consecutive passages in chick embryo chorioallontoic membrane cells. Virus replication was monitored by cytopathic observation, indirect immunoperoxidase, and reverse transcription polymerase chain reaction (RT-PCR). At 72 hours post-infection in third passage, the cytopathic effect was characterized by rounding up of cells, monolayer detachment, intracytoplasmic brownish colouration was readily observed by immunoperoxidase from 24hours p.i in third passage, and at all times the extracted viral RNA from IBV-infected monolayers was demonstrated by RT-PCR. Tissue culture ineffective dose50 (TCID50) was used to measure virus titration performed on primary chick embryo chorioallontoic membrane cells and the titre in twenty passage was 108.6 TCID50/ml. The results obtained in this study suggested that the primary chick embryo chorioallontoic membrane cells can be used for adaptation infectious bronchitis virus (IBV) and may be considered a step forward for the use of these cells in the future for IBV vaccine production

The Effect of Infectious Bronchitis Virus In Embryonated Chicken Eggs

“…The coronavirus causes avian infectious bronchitis (IB), a severe and highly contagious disease of poultry that can cause complex respiratory problems and is among the most wide spread poultry disease (16).The result is characterized by pathological lesions such as dwarfism and congestion in comparison with the control group. Positive RT-PCR samples were inoculated into chicken embryos for virus isolation; no embryos died in the first passage.…”

Clinical and Molecular Study of Canine Enteric Coronavirus in Iraq

“…Other studies reported prevalence ranging from 0.1% [7] to 12% [8]to as high as 26.7% in Japan [9]. Numerous studies were conducted about coronaviruses in Iraq by using the PCR technique in humans [10][11][12][13] and in animals [14][15][16][17][18][19][20]. To the best of our knowledge, no previous studies were conducted about CECoV in Iraq.…”