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IMPROVED METHODS AND CULTURE MEDIA FOR PRODUCTION, QUANTIFICATION AND ISOLATION OF BACTERIOPHAGES

机译:细菌噬菌体生产,定量和分离的改良方法和培养基

摘要

The present invention is directed to a method for producing bacteriophages infecting a bacterial host such as Flavobacterium columnare or Aeromonas sp, the method comprising the steps of adding the bacterium to a sterile culture medium supplemented with mucin or another mucus component, incubating said culture medium, thereby obtaining a bacterial culture, optionally collecting the supernatant containing floating (planktonic) bacterial cells from said culture and transferring the supernatant to a new container in order to discard most of the bio film and adding a bacteriophage infecting the bacterium to said bacterial culture obtained from a previous step, and incubating the culture until the phage yield increases or peaks. The method can also be used for preparing modified enrichment cultures for optimized phage isolation. Addition of mucin to soft-agar culture medium is proposed as an optimized technique for viral titration. This invention also covers all relevant steps in setting up a phage therapy product: phage production, phage quantification and phage isolation.
机译:本发明涉及一种生产感染噬菌体的方法,所述噬菌体感染细菌宿主例如黄杆菌或嗜气单胞菌,所述方法包括将细菌添加到补充有粘蛋白或另一种粘液的无菌培养基中的步骤。成分,孵育所述培养基,从而获得细菌培养物,可选地从所述培养物中收集含有漂浮(浮游性)细菌细胞的上清液,并将上清液转移至新的容器中,以丢弃大部分生物膜并添加感染噬菌体的噬菌体细菌到所述细菌培养物的前一步骤获得,并温育培养物直到噬菌体产量增加或达到峰值。该方法还可以用于制备修饰的富集培养物以优化噬菌体分离。提出将粘蛋白添加到软琼脂培养基中作为病毒滴定的优化技术。本发明还涵盖了建立噬菌体治疗产品的所有相关步骤:噬菌体产生,噬菌体定量和噬菌体分离。

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