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通过对G1P[8]型人野生轮状病毒ZTR-68株的梯度降温传代,筛选适应低温的轮状病毒减毒毒株,为轮状病毒减毒疫苗的制备提供研究基础。自腹泻婴儿粪便样品中分离轮状病毒,在MA104猴肾细胞中自37℃始传代培育,每传代8代后降温3℃,连续传代至28℃,累计传代32代,低温适应过程中检测病毒生物学性状的变化。梯度降温传代后得到的冷适应株RNA基因组电泳迁移条带仍与原始毒株保持一致,血凝滴度降低,病毒蚀斑形态趋于变小且不规则,37℃回复培养时病毒呈现增殖速度减慢且感染性滴度降低的变化。结果显示:野生轮状病毒经冷适应传代后可以得到具有减毒效应的稳定毒株,该毒株出现了部分生物学特征的改变,冷适应株的选育为轮状病毒减毒活疫苗的研制提供了一种备选策略。
The cryogenic rotavirus attenuated strain was screened through the temperature gradient of G1P [8] human wild-type rotavirus ZTR-68, and provided the basis for the preparation of attenuated rotavirus vaccine. Rotavirus was isolated from stool samples from diarrhea infants and subcultured at 37 ℃ in MA104 monkey kidney cells. After 8 passages of each passage, the virus was cooled down to 3 ℃ and continuously passaged to 28 ℃ for 32 passages. Changes in biological traits. The gradient of cold-adapted cold-adapted strains of RNA genome electrophoresis migration bands are still consistent with the original strain, decreased blood clotting titer, virus plaque morphology tends to become smaller and irregular, 37 ℃ when the virus culture showed proliferation rate Slower and less infectious titer changes. The results showed that the stable strain with attenuated effect was obtained after cold-adapted passage of wild-type rotavirus, and some biological characteristics of the strain were observed. The selection of cold-adapted strain was based on the live attenuated rotavirus vaccine Development provides an alternative strategy.