本文選題：分支桿菌噬菌體Leo + 生物學特性　； 參考：《中國人獸共患病學報》2015年03期

【摘要】：目的明確分支桿菌噬菌體Leo的生物學特性及其抗結核作用,為"雞尾酒療法"篩選候選噬菌體。方法采用雙層平板法制備噬菌體Leo,觀察噬菌斑形態(tài);電鏡觀察噬菌體顆粒形態(tài);提取噬菌體DNA,限制性內切酶酶切分析確定其核酸類型;以不同MOI擴增噬菌體Leo,確定最佳MOI及最低MOI;通過一步生長曲線,確定潛伏期及裂解量;采用終點滴定反濁法測定恥垢分支桿菌對噬菌體Leo的耐受突變率;檢測噬菌體Leo對酒精、溫度的耐受性及在不同pH下的裂解能力;通過體外殺結核分枝桿菌實驗,明確噬菌體Leo對結核分枝桿菌的殺滅作用。結果 Leo噬菌斑圓形透明,邊緣清晰,含對稱的多面體頭部,直徑(70±3.0)nm,可彎曲的尾部,尾長(211±31.7)nm;其基因組能被限制性內切酶HindⅢ、BglⅠ切開;最佳MOI為0.0 0001,最低MOI為0.0 001,潛伏期為150min,裂解量74,耐受突變率為10-7。Leo對溫度、酒精敏感,在pH 7.4和pH 5.0時均能裂解宿主菌。Leo作用于結核分支桿菌72h后,Leo組菌量明顯低于空白組(P0.05)。結論分支桿菌噬菌體Leo屬長尾噬菌體科,雙鏈DNA噬菌體,能夠殺滅結核分支桿菌,可作為"雞尾酒療法"的候選噬菌體。
[Abstract]:Objective to identify the biological characteristics and anti tuberculosis effect of Mycobacterium phage Leo, to screen candidate phage for "cocktail therapy". Method using double plate method to prepare phage Leo, observe phage plaque morphology, observe the morphology of phage particles, extract phage DNA, restriction endonuclease digestion analysis to determine its nucleic acid type; The phage Leo was amplified with MOI to determine the best MOI and the lowest MOI; the incubation period and the amount of lysis were determined by one step growth curve. The resistance mutation rate of Mycobacterium phage to phage Leo was determined by the endpoint titration anti turbid method; the resistance of phage Leo to alcohol, temperature and the ability to crack under different pH were detected. Bacteriophage test, clear bacteriophage Leo killing effect of phage on Mycobacterium tuberculosis. Results Leo phage plaque is transparent, edge clear, symmetrical polyhedral head, diameter (70 + 3) nm, curved tail, tail length (211 + 31.7) nm; its genome can be restricted by Hind III, Bgl I incision, the optimum MOI is 0.00001, the lowest MOI is 0.0001, latent MOI is 0.0001, latent MOI is 0.0001, latent MOI is potential 0.0001. The lowest MOI is 0.0001. The lowest MOI is 0.0001. The lowest MOI is 0.0001. The lowest MOI is 0.0001. The lowest MOI is 0.0001, latent MOI is latent and latent MOI is potential 0.0001. Latent MOI is 0.0001, latent MOI potential 0.0001. Latent MOI is 0.0001. Latent MOI potential 0.0001. Latent MOI potential 0.0001. Latent MOI potential 0.0001. Latent MOI potential 0.0001. Latent MOI potential 0.0001. Latent MOI potential 0.0001 The volt period was 150min, the amount of lysis was 74, the tolerance mutation rate was 10-7.Leo to temperature, alcohol sensitive, pH 7.4 and pH 5 could cleave the host bacterium.Leo to act on the Mycobacterium tuberculosis 72h, and the amount of bacteria in Leo group was obviously lower than that of the blank group (P0.05). Conclusion the Mycobacterium phage is a long tail phage, double stranded DNA phage can kill Mycobacterium tuberculosis. It can be used as a candidate phage for "cocktail therapy".
【作者單位】： 重慶醫(yī)科大學附屬第一醫(yī)院呼吸與危重癥醫(yī)學科;成都市第三人民醫(yī)院呼吸內科;重慶市長壽區(qū)人民醫(yī)院呼吸內科;
【基金】：國家“十二五”重大專項課題(2012ZX10003009) 國家臨床重點�？茖ｍ椊涃M資助項目(No.2012-649)資助~~
【分類號】：R378.911

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