MOLECULAR EPIDEMIOLOGY OF ANIMAL-PATHOGENIC STAPHYLOCOCCI,USING PULSED-FIELD GEL ELECTROPHORESIS

使用脉冲场凝胶电泳研究动物致病性葡萄球菌的分子流行病学

• 批准号: 07660419
• 负责人: KAWANO Junichi
• 金额: $ 1.47万
• 依托单位: KOBE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660419/
• 关键词: pulsed-field gel electrophoresis; Staphylococcus; genomic DNA; Staphylococcus hyicus; Staphylococcus intermedius; pathogenicity; パルスフィールド電気泳動

Staphylococcus hyicus and Staphylococcus intermedius have been implicated in a variety of infections in animals, however, both of them are also distributed in healthy animals. For the epidemiologic study of these organisms, an appropriate marker is needed for differentiating virulent and avirulent strains. For that purpose, we investigated whether or not genomic DNA fingerprinting, using pulsed-fiels gel electrophoresis (PFGE), is applicable. First, appropriate conditions of PFGE method was determined, and applied to the analysis of S.hyicus strains from various animals species. Specific banding patterns were observed according to the animal species, suggesting that ecovars based on the genetic background may be present in S.hyicus. Regarding S.hyicus infections in animals, that in pigs is the most serious problem. Most (90%) of strains from diseased pigs showed a specific combination of bands, while strains from healthy ones showed a variety of banding patterns, suggesting that PFGE a … More nalysis may enable us to distinguish virulent strains of S.hyicus. On the other hand, the virulent banding patten was also detected in some strains from healthy pigs. This may suggest that not only in the side of organism but in the side of animal host factors for manifesting disease are present. Exoproteins of S.hyicus strains was analyzed by SDS-PAGE.Virulent strains of S.hyicus revealed an extensive amount of a specific protein, while avirulent ones did remarkablly less amount of the protein or not.With regard to S.intermedius, it showed a high degree of banding patterns in PFGE analysis, suggesting that the pathogenicity could not be assessed by the test. It was proved that strains of this organism are heterogeneous.A mouse model was applied to the analysis of the two species of Staphylococcus. It could not reveal the virulence of strains, suggesting that host factors may be important in pathogenesis of those diseases.We conclude that PFGE analysis is a powerful tool for epidemiologic or ecologic studies of S.hyicus and S.intermedius. Less

葡萄球菌和中间葡萄球菌与多种动物感染有关，但两者也分布在健康动物中。为了对这些生物进行流行病学研究，需要一种合适的标记来区分毒株和无毒株。为此，我们研究了使用脉冲场凝胶电泳（PFGE）的基因组DNA指纹识别是否适用。首先，确定了PFGE法的适宜条件，并将其应用于各种动物的菌种分析。根据动物种类的不同，观察到特定的条带模式，表明基于遗传背景的生态型可能存在。在动物感染中，猪感染是最严重的问题。大多数病猪菌株（90%）表现出特定的条带组合，而健康猪菌株表现出多种条带模式，表明PFGE是一种有效的菌株，进一步的分析可以帮助我们区分猪链球菌的毒力菌株。另一方面，在健康猪的一些毒株中也发现了毒带型。这可能表明，不仅在生物体方面，而且在动物宿主方面也存在表现疾病的因素。采用SDS-PAGE技术对菌种外蛋白进行分析。毒力菌株显示出大量的特定蛋白质，而无毒菌株则显示出显著的蛋白质量较少或没有。中间假丝酵母在PFGE分析中表现出高度的条带模式，表明该菌株的致病性不能通过检测来评估。事实证明，这种生物的菌株是异质的。采用小鼠模型对两种葡萄球菌进行了分析。它不能揭示菌株的毒力，提示宿主因素可能在这些疾病的发病机制中起重要作用。我们认为PFGE分析是研究水蚤和中间蚤流行病学和生态学的有力工具。少

项目成果

Akira Shimizu: "Genomic DNA fingerprinting,using pulsed-field gel electrophoresis,of Staphylococcus intermedius isolated from dogs" American Journal of Veterinary Research. 57・10. 1458-1462 (1996)

Akira Shimizu：“使用脉冲场凝胶电泳对从狗中分离出的中间葡萄球菌进行基因组 DNA 指纹分析”，美国兽医研究杂志 57・10（1996 年）。

Akira Shimizu: "Genomic DNA fingerprinting, using pulsed-field gel electrophoresis, of Staphylococcus intermedivs isolated from dogs" American Journal of Veterinary Research. 57. 1458-1462 (1996)

Akira Shimizu：“使用脉冲场凝胶电泳对从狗身上分离出的中间葡萄球菌进行基因组 DNA 指纹分析”《美国兽医研究杂志》。

Akira Shimizu: "Pulsed-field gel electrophoresis of Staphylococcus hyicus and Staphylococcus chromogenes genomic DNA and its taxonomic,epidemiologic and ecologic applications in veterinary medicine" Journal of Veterinary Medical Science. (印刷中).

Akira Shimizu：“猪葡萄球菌和产色葡萄球菌基因组 DNA 的脉冲场凝胶电泳及其在兽医学中的分类学、流行病学和生态学应用”《兽医医学科学杂志》（正在出版）。

AKIRA SHIMIZU: "Genomic DNA fingerprinting, using pulsed-field gel electrophoresis, of Staphylococcus intermedius isolated from dogs" American Journal of Veterinary Research. Vol.57-No.10. 1458-1462 (1996)

AKIRA SHIMIZU：“使用脉冲场凝胶电泳对从狗身上分离出的中间葡萄球菌进行基因组 DNA 指纹分析”《美国兽医研究杂志》。

Akira Shimizu: "Pulsed-field gel electrophoresis of Staphylococcus hyicus and Staphylococcus chromogenes genomic DNA and its taxonomic epidemiologic and ecologic applications in veterinary medicine" Journal of Veterinary Medical Science. (印刷中).

Akira Shimizu：“猪葡萄球菌和产色葡萄球菌基因组 DNA 的脉冲场凝胶电泳及其在兽医学中的分类流行病学和生态学应用”《兽医医学科学》杂志（正在出版）。