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Helicobacter Pylori--Nutrition, Serum and Culturability

幽门螺杆菌——营养、血清和可培养性

基本信息

批准号：
6635382
负责人：
TRACI L TESTERMAN
金额：
$ 4.81万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-06-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6635382
关键词：
Helicobacter bacterial genetics feces gene induction /repression growth media intestines microorganism culture microorganism growth saliva transposon /insertion element

项目摘要

DESCRIPTION (provided by the applicant): Helicobacter pylori is a fastidious, microaerophilic organism strongly associated with peptic ulcers and gastric carcinoma. Growth requirements of this organism have only been partially defined, and isolation of H. pylori from in vivo sites and the environment remains challenging. H. pylori normally requires blood or serum for growth in vitro. Our preliminary experiments provide insights into growth and serum requirements and reveal that recovery of H. pylori from frozen stocks is significantly improved through the use of a partially defined medium rather than standard complex media. We hypothesize that the use of complex media inhibits recovery of dormant or oxidatively stressed organisms, precluding isolation of H. pylori from environmental sources. We propose to extend our preliminary studies by further defining growth requirements and identifying the component(s) of serum most responsible for growth augmentation. A transposon library of H. pylori mutants will be screened to identify clones which fail to grow in defined medium without serum. Since serum both augments growth and prolongs culture viability, genes identified in this screen are expected to provide clues regarding serum's function in H. pylori growth. These genes may also play a role in stationary phase survival of H. pylori and may impact virulence and transmission of the organism as well. The optimized medium will be used in conjunction with other measurements of viability to evaluate extragastric survival of H. pylori from sites such as the intestine, saliva, and feces. Results of these studies should extend the understanding of H. pylori growth requirements and the genes involved, particularly requirements relevant to recovery of the organism from various environmental conditions. This research will provide more useful media formulations for both clinical and research applications, and may also be applicable to the culture of other fastidious bacteria.

描述（申请人提供）：幽门螺杆菌是一种挑剔的、嗜微气的微生物，与消化性溃疡和胃癌密切相关。这种生物的生长需求只是部分定义，从体内和环境中分离幽门螺杆菌仍然具有挑战性。幽门螺旋杆菌通常需要血液或血清才能在体外生长。我们的初步实验提供了对生长和血清需求的见解，并揭示了通过使用部分定义的培养基而不是标准的复杂培养基，从冷冻储存液中回收幽门螺杆菌的效果显著提高。我们假设，使用复杂的培养基抑制休眠或氧化应激生物体的恢复，排除从环境源分离幽门螺杆菌。我们建议通过进一步确定生长需求和确定血清中最能促进生长的成分来扩展我们的初步研究。将筛选幽门螺杆菌突变体的转座子文库，以鉴定在没有血清的特定培养基中不能生长的克隆。由于血清既能促进生长又能延长培养活力，因此在该筛选中鉴定的基因有望为血清在幽门螺杆菌生长中的功能提供线索。这些基因也可能在幽门螺杆菌的静止期存活中发挥作用，并可能影响该生物的毒力和传播。优化的培养基将与其他生存能力测量一起使用，以评估来自肠道、唾液和粪便等部位的幽门螺杆菌的胃外存活。这些研究的结果应该扩展对幽门螺杆菌生长需求和相关基因的理解，特别是与有机体从各种环境条件中恢复相关的需求。本研究将为临床和研究应用提供更多有用的培养基配方，也可能适用于其他挑剔细菌的培养。