目的:比较头孢硝噻吩液体(Cefinase)、头孢硝噻吩纸片(Dryslide)、三叶草实验(Cloverleaf)、青霉素边缘实验(Penicillin disk zone edge)检测金黄色葡萄球菌β-内酰胺酶的敏感性和特异性。方法:分别用Cefinase,Dryslide,Cloverleaf,Peni...目的:比较头孢硝噻吩液体(Cefinase)、头孢硝噻吩纸片(Dryslide)、三叶草实验(Cloverleaf)、青霉素边缘实验(Penicillin disk zone edge)检测金黄色葡萄球菌β-内酰胺酶的敏感性和特异性。方法:分别用Cefinase,Dryslide,Cloverleaf,Penicillin disk zone edge检测金黄色葡萄球菌β-内酰胺酶。结果:4种方法敏感度分别为83%、66%、91%、91%。特异度分别为100%、97%、100%、100%。结论:4种方法在检测应用上都能为临床提供可靠的结果,但Cloverleaf和Penicillin disk zone edge检测β-内酰胺酶的试验较敏感,在操作上用头孢硝噻吩纸片法更能被实验室所接受。展开更多
We developed a colorimetric assay to quantify clavulanic acid （CA） in culture broth of Streptomyces clavuligerus, to facilitate screening of a large number of S. clavuligerus mutants. The assay is based on a β-1act...We developed a colorimetric assay to quantify clavulanic acid （CA） in culture broth of Streptomyces clavuligerus, to facilitate screening of a large number of S. clavuligerus mutants. The assay is based on a β-1actamase-catalyzed reaction, in which the yellow substrate nitrocefin （λmax=390 nm） is converted to a red product （λmax=486 nm）. Since CA can irreversibly inhibit β-1actamase activity, the level of CA in a sample can be measured as a function of the A390]A486 ratio in the assay mixture. The sensitivity and detection window of the assay were determined to be 50 μg L -1 and 50 μg L to 10 mg L-1, respectively. The reliability of the assay was confirmed by comparing assay results with those obtained by HPLC. The assay was used to screen a pool of 65 S. clavuligerus mutants and was reliable for identifying CA over-producing mutants. Therefore, the assay saves time and labor in large-scale mutant screening and evaluation tasks. The detection window and the reliability of this assay are markedly better than those of previously reported CA assays. This assay method is suitable for high throughput screening of microbial samples and allows direct visual observation of CA levels on agar plates.展开更多
文摘目的:比较头孢硝噻吩液体(Cefinase)、头孢硝噻吩纸片(Dryslide)、三叶草实验(Cloverleaf)、青霉素边缘实验(Penicillin disk zone edge)检测金黄色葡萄球菌β-内酰胺酶的敏感性和特异性。方法:分别用Cefinase,Dryslide,Cloverleaf,Penicillin disk zone edge检测金黄色葡萄球菌β-内酰胺酶。结果:4种方法敏感度分别为83%、66%、91%、91%。特异度分别为100%、97%、100%、100%。结论:4种方法在检测应用上都能为临床提供可靠的结果,但Cloverleaf和Penicillin disk zone edge检测β-内酰胺酶的试验较敏感,在操作上用头孢硝噻吩纸片法更能被实验室所接受。
基金supported by the Young Scientists Fund (Grant No. 31000025) from the National Natural Science Foundation of ChinaNational High Technology Research and Development Program of China (Grant No. 2012AA021302)
文摘We developed a colorimetric assay to quantify clavulanic acid （CA） in culture broth of Streptomyces clavuligerus, to facilitate screening of a large number of S. clavuligerus mutants. The assay is based on a β-1actamase-catalyzed reaction, in which the yellow substrate nitrocefin （λmax=390 nm） is converted to a red product （λmax=486 nm）. Since CA can irreversibly inhibit β-1actamase activity, the level of CA in a sample can be measured as a function of the A390]A486 ratio in the assay mixture. The sensitivity and detection window of the assay were determined to be 50 μg L -1 and 50 μg L to 10 mg L-1, respectively. The reliability of the assay was confirmed by comparing assay results with those obtained by HPLC. The assay was used to screen a pool of 65 S. clavuligerus mutants and was reliable for identifying CA over-producing mutants. Therefore, the assay saves time and labor in large-scale mutant screening and evaluation tasks. The detection window and the reliability of this assay are markedly better than those of previously reported CA assays. This assay method is suitable for high throughput screening of microbial samples and allows direct visual observation of CA levels on agar plates.
