Development of optimized protocol for culturing African swine fever virus field isolates in MA104 cells.

IF 1.1 4区 农林科学 Q2 Veterinary Canadian journal of veterinary research = Revue canadienne de recherche veterinaire Pub Date : 2022-10-01
Hyeok-Il Kwon, Duy Tien Do, Hung Van Vo, Seung-Chul Lee, Min Ho Kim, Dung Thi Thuy Nguyen, Tan Minh Tran, Quang Tin Vinh Le, Tram Thi Ngoc Ngo, Nam Minh Nguyen, Joo Young Lee, Toan Tat Nguyen
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Abstract

The goal of this study was to identify a candidate commercial cell line for the replication of African swine fever virus (ASFV) by comparing several available cell lines with various medium factors. In the sensitivity test of cells, MA104 and MARC-145 had strong potential for ASFV replication. Next, MA104 cells were used to compare the adaptation of ASFV obtained from tissue homogenates and blood samples in various infectious media. At the 10th passage, the ASFV obtained from the blood sample had a significantly higher viral load than that obtained from the tissue sample (P = 0.000), exhibiting a mean cycle threshold (Ct) value = 20.39 ± 1.99 compared with 25.36 ± 2.11. For blood samples, ASFV grew on infectious medium B more robustly than on infectious medium A (P = 0.006), corresponding to a Ct value = 19.58 ± 2.10 versus 21.20 ± 1.47. African swine fever virus originating from blood specimens continued to multiply gradually and peaked in the 15th passage, exhibiting a Ct value = 14.36 ± 0.22 in infectious medium B and a Ct value = 15.42 ± 0.14 in infectious medium A. When ASFV was cultured from tissue homogenates, however, there was no difference (P = 0.062) in ASFV growth between infectious media A and B. A model was developed to enhance ASFV replication through adaptation to MA104 cells. The lack of mutation at the genetic segments encoding p72, p54, p30, and the central hypervariable region (CVR) in serial culture passages is important in increasing the probability of maintaining immunogenicity when developing a vaccine candidate.

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MA104细胞培养非洲猪瘟病毒分离株优化方案的建立。
本研究的目的是通过比较几种具有不同培养基因素的可用细胞系,确定一种用于复制非洲猪瘟病毒(ASFV)的候选商业细胞系。在细胞敏感性试验中,MA104和MARC-145具有很强的ASFV复制潜力。接下来,使用MA104细胞比较从组织匀浆和血液样本中获得的ASFV在各种感染介质中的适应性。在第10代时,从血液样本中获得的ASFV病毒载量明显高于从组织样本中获得的病毒载量(P = 0.000),平均周期阈值(Ct)值为20.39±1.99,而从组织样本中获得的病毒载量为25.36±2.11。在血液样本中,ASFV在感染培养基B上比在感染培养基A上生长更强劲(P = 0.006),对应的Ct值分别为19.58±2.10和21.20±1.47。来自血液标本的非洲猪瘟病毒继续逐渐繁殖,并在第15代达到顶峰,在感染培养基B中Ct值为14.36±0.22,在感染培养基a中Ct值为15.42±0.14,而在组织培养液中培养ASFV时,ASFV在感染培养基a和B之间的生长没有差异(P = 0.062)。在连续培养传代中,编码p72、p54、p30和中央高变区(CVR)的基因片段缺乏突变,在开发候选疫苗时增加维持免疫原性的可能性是重要的。
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来源期刊
CiteScore
1.80
自引率
0.00%
发文量
58
审稿时长
>24 weeks
期刊介绍: The Canadian Journal of Veterinary Research, published by the Canadian Veterinary Medical Association, is Canada''s only veterinary research publication. This quarterly peer-reviewed online-only journal has earned a wide international readership through the publishing of high quality scientific papers in the field of veterinary medicine. The Journal publishes the results of original research in veterinary and comparative medicine.
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