Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line
作者： Nafi’u Lawal , Mohd Hair-Bejo , Siti Suri Arshad , Abdul Rahman Omar , Aini Ideris
DOI: 10.1155/2017/8359047
关键词:
摘要: Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted propagated BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) subtly observed at P1 both isolates 72 hours postinoculation (pi). CPE became prominent P5 with rounding, cytoplasmic vacuoles, granulation, detachment flask starting day 3 pi, 7 days pi titers of 109.50 TCID50/mL log109.80 TCID50/mL for UPM190, respectively. subtle P17 disappeared P18 P19 However, the presence IBDV confirmed immunoperoxidase, immunofluorescence, RT-PCR techniques. Phylogenetic analysis showed that these two vvIBDV. It appears a single mutation D279N could facilitate vvIBDV strain adaptability CEE cultures.
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