Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that represents a major problem in many hospitals because of its increased resistance to antibiotics and the ability to cause nosocomial ...Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that represents a major problem in many hospitals because of its increased resistance to antibiotics and the ability to cause nosocomial infections. The present study aimed to phenotype and genotype isolates of P. aeruginosa from inpatients with UTIs at Urology and Nephrology center, Mansoura, Egypt to study their relatedness. Methods: Thirty nine isolates of P. aeruginosa were phenotypically typed by determination of O-serotypes by slide agglutination technique and antimicrobial resistance patterns by disk-diffusion method. The genetic diversity of isolates was illustrated by performing RAPD-PCR using M13 primer. Results: Serotypes O11, O6 and O10 were the most prevalent. Isolates showed high resistance rates to antipseudmonal antibiotics with high incidence (51.3%) of multidrug resistance (MDR). Amikacin was the most effective. A significant correlation was found between O6, O10 and MDR. A relatively high polymorphism was demonstrated among P. aeruginosa isolates by using RAPD-M13 fingerprinting. Cross transmission was suggested by phenotypically and clonally identical isolates. Conclusion: The study demonstrates the role of combining both classical and molecular typing as a valuable mean to study the origin and cross transmission of P. aeruginosa in UTIs for better assessment of treatment and infection control.展开更多
文摘Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that represents a major problem in many hospitals because of its increased resistance to antibiotics and the ability to cause nosocomial infections. The present study aimed to phenotype and genotype isolates of P. aeruginosa from inpatients with UTIs at Urology and Nephrology center, Mansoura, Egypt to study their relatedness. Methods: Thirty nine isolates of P. aeruginosa were phenotypically typed by determination of O-serotypes by slide agglutination technique and antimicrobial resistance patterns by disk-diffusion method. The genetic diversity of isolates was illustrated by performing RAPD-PCR using M13 primer. Results: Serotypes O11, O6 and O10 were the most prevalent. Isolates showed high resistance rates to antipseudmonal antibiotics with high incidence (51.3%) of multidrug resistance (MDR). Amikacin was the most effective. A significant correlation was found between O6, O10 and MDR. A relatively high polymorphism was demonstrated among P. aeruginosa isolates by using RAPD-M13 fingerprinting. Cross transmission was suggested by phenotypically and clonally identical isolates. Conclusion: The study demonstrates the role of combining both classical and molecular typing as a valuable mean to study the origin and cross transmission of P. aeruginosa in UTIs for better assessment of treatment and infection control.
文摘为构建含有海肾萤光素酶(Rluc)报告基因的O型口蹄疫病毒(FMDV)亚基因组复制子,本研究在前期构建的O型FMDV cDNA感染性克隆的基础上,利用融合PCR的方法,以Rluc报告基因替换FMDV的前导蛋白Lb基因和结构蛋白P1基因,构建含有Rluc报告基因的FMDV亚基因组复制子FMDVRep。将该复制子质粒线性化后,经体外转录制备复制子全长RNA,转染BHK-21细胞,通过RT-PCR方法和间接免疫荧光试验分别检测该复制子RNA的自主复制能力以及FMDV非结构蛋白3B的表达情况。结果显示,复制子RNA在BHK-21细胞中能够进行自主复制并且能够表达病毒的非结构蛋白。Rluc活性检测结果表明,该复制子RNA在BHK-21细胞中可以高效表达Rluc,在转染后2 h^12 h Rluc活性呈线性递增,进一步反映该复制子具有良好的表达外源基因的能力。O型FMDV亚基因组复制子的构建,为深入研究FMDV的复制和翻译机制奠定了基础。
