Background and Prupose: Antibiotic resistance is a major global health concern. In addition to the existing data on the prevalence of bacterial resistance to antibiotics, there are patchy data on bacterial resistance ...Background and Prupose: Antibiotic resistance is a major global health concern. In addition to the existing data on the prevalence of bacterial resistance to antibiotics, there are patchy data on bacterial resistance to aminoglycosides in Burkina Faso. In this study, we determined the prevalence of aminoglycoside resistance genes in E. coli, including aac(3)-IIc, aac(6)-Ib and armA in Ouagadougou, and determined which antibiotics in this class are most affected by resistance. Material and Methods: This study was conducted on 216 E. coli strains collected from the biomedical analysis laboratories of Saint Camille and Schiphra hospitals. E. coli strains were isolated from pus and urine samples collected between September 2018 and January 2019. Antibiotic susceptibility testing was performed using aminoglycosides, β-lactams, fluoroquinolones, and sulfonamides. Aminoglycoside resistance genes were detected in strains with at least one aminoglycoside resistance gene using conventional/multiplex PCR. Results: Aminoglycoside resistance was observed in 46.8% (101/216) of strains. The resistance rates were respectively 45.37% for Tobramycin, 32.40% for Gentamicin, 14.81% for Kanamycin, 2.31% for Netilmicin, 1.84% for Neomycin, and 0.46% for Amikacin. PCR showed that 86 strains (85.15%) possessed the aac(3)-IIc gene, 71 strains or 70.30%) possessed the aac(6’)-Ib gene, and nine strains (8.91%) possessed the armA gene. Conclusion: Aminoglycoside resistance in pathogenic E. coli strains is mainly due to the presence of the aac(3’)-IIc and aac(6’)-Ib genes. The presence of armA was first reported in Burkina Faso. Netilmicin, Neomycin and Amikacin are good therapeutic options for treating urinary tract and pus-forming infections.展开更多
以E.coliDH5α基因组为模板PCR扩增得到无信号肽的碱性磷酸酶基因片段,与胞内融合表达型T载体连接得到重组质粒,转化表达宿主E.coliBL21(DE3)和E.coliorigami(DE3),经0.1 mM IPTG诱导表达,超声破碎细胞后,SDS-PAGE分析可溶性,融合蛋白在...以E.coliDH5α基因组为模板PCR扩增得到无信号肽的碱性磷酸酶基因片段,与胞内融合表达型T载体连接得到重组质粒,转化表达宿主E.coliBL21(DE3)和E.coliorigami(DE3),经0.1 mM IPTG诱导表达,超声破碎细胞后,SDS-PAGE分析可溶性,融合蛋白在E.coliorigami(DE3)中的可溶蛋白含量比E.coliBL21(DE3)中高.融合蛋白的可溶部分经Ni2+螯合亲和纯化,纯化后E.coliorigami(DE3)中蛋白活性比E.coliBL21(DE3)中明显提高,达到1 614.3U/mg蛋白.展开更多
本文继大肠杆菌表达含凝血酶切点的人重组IL-3融合蛋白成功的基础上进一步探讨了天然型rhIL-3的纯化。超声破碎细菌细胞得包涵体,经洗涤处理可使包涵体纯度达90％,用8mol/L尿素变性溶解包涵体沉淀后直接稀释复性,再超滤浓缩、凝血酶消化...本文继大肠杆菌表达含凝血酶切点的人重组IL-3融合蛋白成功的基础上进一步探讨了天然型rhIL-3的纯化。超声破碎细菌细胞得包涵体,经洗涤处理可使包涵体纯度达90％,用8mol/L尿素变性溶解包涵体沉淀后直接稀释复性,再超滤浓缩、凝血酶消化,释放天然型rhIL-3。经DEAE Sepharose Fast Flow和Sepharcyl-100 HR层析得到天然型IL-3,纯度达96％,回收率20％以上,具有刺激正常人骨髓细胞形成集落的活性。本实验为大批量重组IL-3的生产创造了条件。展开更多
文摘Background and Prupose: Antibiotic resistance is a major global health concern. In addition to the existing data on the prevalence of bacterial resistance to antibiotics, there are patchy data on bacterial resistance to aminoglycosides in Burkina Faso. In this study, we determined the prevalence of aminoglycoside resistance genes in E. coli, including aac(3)-IIc, aac(6)-Ib and armA in Ouagadougou, and determined which antibiotics in this class are most affected by resistance. Material and Methods: This study was conducted on 216 E. coli strains collected from the biomedical analysis laboratories of Saint Camille and Schiphra hospitals. E. coli strains were isolated from pus and urine samples collected between September 2018 and January 2019. Antibiotic susceptibility testing was performed using aminoglycosides, β-lactams, fluoroquinolones, and sulfonamides. Aminoglycoside resistance genes were detected in strains with at least one aminoglycoside resistance gene using conventional/multiplex PCR. Results: Aminoglycoside resistance was observed in 46.8% (101/216) of strains. The resistance rates were respectively 45.37% for Tobramycin, 32.40% for Gentamicin, 14.81% for Kanamycin, 2.31% for Netilmicin, 1.84% for Neomycin, and 0.46% for Amikacin. PCR showed that 86 strains (85.15%) possessed the aac(3)-IIc gene, 71 strains or 70.30%) possessed the aac(6’)-Ib gene, and nine strains (8.91%) possessed the armA gene. Conclusion: Aminoglycoside resistance in pathogenic E. coli strains is mainly due to the presence of the aac(3’)-IIc and aac(6’)-Ib genes. The presence of armA was first reported in Burkina Faso. Netilmicin, Neomycin and Amikacin are good therapeutic options for treating urinary tract and pus-forming infections.
文摘以E.coliDH5α基因组为模板PCR扩增得到无信号肽的碱性磷酸酶基因片段,与胞内融合表达型T载体连接得到重组质粒,转化表达宿主E.coliBL21(DE3)和E.coliorigami(DE3),经0.1 mM IPTG诱导表达,超声破碎细胞后,SDS-PAGE分析可溶性,融合蛋白在E.coliorigami(DE3)中的可溶蛋白含量比E.coliBL21(DE3)中高.融合蛋白的可溶部分经Ni2+螯合亲和纯化,纯化后E.coliorigami(DE3)中蛋白活性比E.coliBL21(DE3)中明显提高,达到1 614.3U/mg蛋白.
文摘本文继大肠杆菌表达含凝血酶切点的人重组IL-3融合蛋白成功的基础上进一步探讨了天然型rhIL-3的纯化。超声破碎细菌细胞得包涵体,经洗涤处理可使包涵体纯度达90％,用8mol/L尿素变性溶解包涵体沉淀后直接稀释复性,再超滤浓缩、凝血酶消化,释放天然型rhIL-3。经DEAE Sepharose Fast Flow和Sepharcyl-100 HR层析得到天然型IL-3,纯度达96％,回收率20％以上,具有刺激正常人骨髓细胞形成集落的活性。本实验为大批量重组IL-3的生产创造了条件。
