噬菌体phiYe-F10的裂解谱的测定以及裂解能力与宿主毒力基因间关系分析

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目的为测定小肠结肠炎耶尔森菌噬菌体phiYe-F10的裂解谱,分析噬菌体phiYe-F10裂解能力与宿主毒力基因间的关系。方法采用双层平板法观察噬菌体phiYe-F10对213株小肠结肠炎耶尔森菌,36株假结核耶尔森菌和1株鼠疫耶尔森菌的裂解能力;同时对不同来源不同地区的小肠耶尔森菌进行黏附侵袭位点基因(ail)、肠毒素基因(ystA、ystB)、黏附素基因A(yadA)、毒力因子基因F(virF)、O∶3血清型特异性基因(rfbc)检测和血清型别鉴定。结果表明213株小肠结肠炎耶尔森菌包括O∶3血清型84株(其中rfbc+78株),O∶5血清型10株,O∶8血清型13株,O∶9血清型34株,其它及未分型的共72株。携带毒力质粒的典型致病性小肠耶尔森菌(ail+,ystA+,ystB-,yadA+,virF+)77株,毒力质粒丢失的致病性小肠结肠炎耶尔森菌(ail+、ystA+、ystB-、yadA-、virF-)15株,其它非致病基因型121株。噬菌体phiYe-F10裂解71株O∶3小肠结肠炎耶尔森氏菌,其中致病性小肠结肠炎耶尔森菌52株,非致病性小肠结肠炎耶尔森菌19株。对O∶5,O∶9血清型和其它菌株均不裂解,对O∶3的裂解率高达84.5%(71/84)。噬菌体phiYe-F10对假结核耶尔森菌和鼠疫耶尔森氏菌均不裂解。结论:phiYe-F10噬菌体是一株小肠结肠炎耶尔森菌的裂性噬菌体,在25℃时表现出一个典型的窄裂解谱,高度专一性裂解O∶3血清型小肠结肠炎耶尔森菌。phiYe-F10对典型致病性小肠结肠炎耶尔森菌的裂解能力明显高于非致病性的小肠结肠炎耶尔森菌 The purpose of this study was to determine the lytic spectrum of phage phiYe-F10 of enterocolitis Yersinia enterocolitica, and to analyze the relationship between phiYe-F10 lytic capacity of phage and host virulence genes. Methods The phage phiYe-F10 was used to detect the lytic capacity of 213 strains of Yersinia enterocolitica, 36 strains of Yersinia pseudotuberculosis and 1 strain of Y. pestis by double-layer plate method. Meanwhile, Yersinia enterocolitica was used to detect the expression of ail, ystA and ystB, yadA, virF, O: 3 serotype specific gene (rfbc ) Testing and serological type identification. The results showed that 213 Yersinia enterocolitica strains including O: 3 serotype 84 (of which rfbc + 78), O: 5 serotype 10, O: 8 serotype 13, O: 9 serotype 34 , Other and undifferentiated a total of 72 strains. 77 pathogenic Yersinia enterocolitica (ail +, ystB +, yadA +, virF +) carrying virulence plasmids, Yersinia enteritidis (ail +, ystA +, ystB -, yadA-, virF-) 15 strains, 121 other non-pathogenic genotypes. The bacteriophage phiYe-F10 lysed 71 strains of O: 3 Yersinia enterocolitica, including 52 pathogenic Yersinia enterocolitica and 19 Yersinia enteritidis. No O: 5, O: 9 serotypes and other strains were cleaved, the cleavage rate of O: 3 was as high as 84.5% (71/84). Phage phiYe-F10 did not lyse against Yersinia pestis and Y. pestis. Conclusions: The phiYe-F10 phage is a fission phage from Yersinia enterocolitica exhibiting a typical narrow cleavage profile at 25 ° C and highly specific cleavage of O: 3 serotypes of enterocolitis Yersinia bacteria. The lytic capacity of phiYe-F10 against Yersinia enterocolitica was significantly higher than that of non-pathogenic Yersinia enterocolitica
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