Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line.

IF 1.1 Q4 VIROLOGY
Advances in Virology Pub Date : 2017-01-01 Epub Date: 2017-11-05 DOI:10.1155/2017/8359047
Nafi'u Lawal, Mohd Hair-Bejo, Siti Suri Arshad, Abdul Rahman Omar, Aini Ideris
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引用次数: 14

Abstract

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and 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 and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL and log109.80 TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures.

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两株马来西亚非常强毒传染性法氏囊病病毒在BGM-70细胞系上的适应和分子特性
将从当地IBD暴发中分离到的2株马来西亚非常强毒传染性法氏囊病病毒(vvIBDV) UPM0081和UPM190(分别称为UPMB00/81和UPM04/190)通过绒毛尿囊膜(CAM)途径在特异性无病原体(SPF)鸡胚蛋(CEE)中连续传代12次(EP12)。EP12分离物进一步适应并在BGM-70细胞系中连续繁殖达20代(P20)。在接种后72小时(pi),两个分离株在P1处观察到特征性细胞病变效应(cpe)。UPM0081和UPM190的滴度分别为109.50 TCID50/mL和log109.80 TCID50/mL,从第3天开始到第7天,CPE在P5时变得突出,细胞变圆,细胞质空泡,肉芽形成,从瓶中脱落。UPM0081和UPM190的CPE在P17处变弱,在P18和P19处消失。然而,通过免疫过氧化物酶、免疫荧光和RT-PCR技术证实了IBDV的存在。系统发育分析表明,这两个分离株均属于vvIBDV。UPM190和UPM0081 IBDV分离株在D279N的单一突变似乎可以促进vvIBDV菌株在CEE和BGM-70培养中的适应性。
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来源期刊
CiteScore
2.30
自引率
0.00%
发文量
23
审稿时长
22 weeks
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