PLAQUE-FORMATION BY VIRULENT SHIGELLA-FLEXNERI
PLAQUE-FORMATION BY VIRULENT SHIGELLA-FLEXNERI
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DOI：
10.1128/iai.48.1.124-129.1985
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发表时间：
1985-01-01
影响因子：
3.1
通讯作者：
FORMAL, SB
中科院分区：
文献类型：
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作者：
OAKS, EV;WINGFIELD, ME;FORMAL, SB
在体外组织培养空斑试验，以研究细胞内复制和细胞间传播的毒性志贺氏菌空斑形成在HeLa细胞单层的琼脂糖覆盖含有组织培养基和庆大霉素，消除细胞外细菌生长的存在下。在显微镜下，空斑的特征是中心区域的死亡宿主细胞被感染志贺氏菌的细胞包围。远离空斑中心的细胞未被感染。氯霉素或萘啶酸的重叠完全废除了斑块的形成。当感染的单层从37 °移动到30 °时，噬斑形成被完全抑制。C.将感染的单层从30 ℃移动到30 ℃。C，其中不形成斑块，至37 ℃。C导致斑块的形成。鼠伤寒沙门氏boydii，S. sonnei（Ⅰ型）和所有6种血清型的S.弗氏产生斑块。从菌斑中分离的志贺菌为Sereny试验阳性，含有140兆道尔顿质粒，对庆大霉素敏感。非侵袭性志贺菌不形成斑块。
An in vitro tissue culture plaque assay was developed to investigate the intracellular replication and intercellular spread of virulent shigellae plaques were formed in HeLa cell monolayers in the presence of an agarose overlay containing tissue culture medium and gentamicin, which eliminated extracellular bacterial growth. Microscopically, the plaques were characterized by a central area of dead host cells surrounded by cells infected with shigellae. Cells further away from the plaque center were uninfected. Inclusion of chloroamphenicol or nalidixic acid in the overlap completely abolished plaque formation. Plaque formation was completely inhibited when infected monolayers were shifted from 37 to 30.degree. C. Shifting infected monolayers from 30.degree. C, where plaques do not form, to 37.degree. C resulted in the formation of plaques. Cultures of S. boydii, S. sonnei (form I), and all 6 serotypes of S. flexneri produced plaques. Shigellae isolated from plaques were Sereny test positive, contained a 140 megadalton plasmid, and were gentamicin sensitive. Noninvasive shigellae did not form plaques.