Propagation and Molecular Characterization of Bioreactor Adapted Very Virulent Infectious Bursal Disease Virus Isolates of Malaysia.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2018-09-02 eCollection Date: 2018-01-01 DOI:10.1155/2018/1068758

Nafi'u Lawal, Mohd Hair-Bejo, Siti Suri Arshad, Abdul Rahman Omar, Aini Ideris

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引用次数: 4

Abstract

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 (also known as B00/81) and UPM190 (also known as UPM04/190) isolated from local IBD outbreaks in 2000 and 2004, respectively, were separately passaged for 12 consecutive times in 11-day-old specific pathogen free (SPF) chicken embryonated eggs (CEE) via the chorioallantoic membrane (CAM) route. The CEE passage 8 (EP8) isolates were passaged once in BGM-70 cell line yielding UPM0081EP8BGMP1 and UPM190EP8BGMP1, while the EP12 isolates were passaged 15 times in BGM-70 cell line yielding UPM0081EP12BGMP15 and UPM190EP12BGMP15 using T25 tissue culture flask. These isolates were all propagated once in bioreactor using cytodex 1 as microcarrier at 3 g per liter (3 g/L) yielding UPM0081EP8BGMP1BP1, UPM190EP8BGMP1BP1, UPM0081EP12BGMP15BP1, and UPM190EP12BGMP15BP1 isolates. The viruses were harvested at 3 days after inoculation, following the appearance of cytopathic effects (CPE) characterized by detachment from the microcarrier using standard protocol and filtered using 0.2 μm syringe filter. The filtrates were positive for IBDV by RT-PCR and immunofluorescence. Sequence and phylogenetic tree analysis indicated that the isolates were of the vvIBDV strains and were not different from the flask propagated parental viruses.

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马来西亚非常强毒传染性法氏囊病病毒分离株的生物反应器适应性繁殖和分子特性。

分别从2000年和2004年当地IBD暴发中分离出两株马来西亚非常强毒的传染性法氏囊病病毒(vvIBDV) UPM0081(也称为B00/81)和UPM190(也称为UPM04/190)，分别通过绒毛尿囊膜(CAM)途径在11日龄的无特异性病原体(SPF)鸡胚蛋(CEE)中连续传代12次。CEE传代8 (EP8)分离物在BGM-70细胞系中传代1次，产生UPM0081EP8BGMP1和UPM190EP8BGMP1, EP12分离物在BGM-70细胞系中传代15次，产生UPM0081EP12BGMP15和UPM190EP12BGMP15，使用T25组织培养瓶。这些分离株均在生物反应器中以3 g/L (3 g/L)的cytodex 1为微载体繁殖一次，获得UPM0081EP8BGMP1BP1、UPM190EP8BGMP1BP1、UPM0081EP12BGMP15BP1和UPM190EP12BGMP15BP1分离株。接种后3天，病毒出现细胞病变效应(CPE)，采用标准方案从微载体分离，并用0.2 μm注射器过滤器过滤。RT-PCR和免疫荧光检测结果均为IBDV阳性。序列分析和系统进化树分析表明，分离株为vvIBDV株，与烧瓶繁殖的亲本病毒无明显差异。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.