噬菌体的检查及效价测定（Detection and titer determination of bacteriophage）.docVIP

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噬菌体的检查及效价测定（Detection and titer determination of bacteriophage）.doc

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噬菌体的检查及效价测定（Detection and titer determination of bacteriophage）

噬菌体的检查及效价测定（Detection and titer determination of bacteriophage） 1, purpose: 1.1 to understand the meaning of phage titer and the principle of its determination. 1.2 learn how to check bacteriophages. 1.3 to master the operational skills of double layer agar plate method for the determination of phage titer. 2, principle: Bacteriophage is a kind of virus that is specialized for microorganisms such as bacteria and actinomyces. Its individual shape is extremely small, and it can not be measured by routine microbial counting method. When a phage infection bacteria will cause cracking sensitive bacteria rapidly, the release of large amounts of progeny phage, which then spread and infection of the surrounding cells, the suspension containing sensitive bacteria from turbidity gradually becomes clear, or the naked eye can see the plaque - plaque in the tablet containing sensitive bacteria. Understanding the characteristics of bacteriophages, rapid detection, isolation, and determination of potency are important for preventing phage contamination in production and scientific research. The sample can be fermented liquid, air, sewage, soil, etc. (as for the object that cannot be sampled and examined, cotton soaked with sterile water can be applied to the surface as a check sample). In order to isolate, the number of phages in the sample can be increased by multiplication. Phage detection using biological assays requires about 12h, so that phage contamination can not be determined in a timely manner. Rapid detection can be used to determine if phage contamination is necessary to take necessary control measures. According to the normal fermentation (Culture) liquid after centrifugation was little precipitation, supernatant protein content, after heating is still clear and infected with phage; fermentation (Culture) active protein liquid after centrifugation, the supernatant containing self cleaving bacteria escape, protein denaturation occurs after heating, and in the light appe