图4. BacTiter-Glo™ 检测与典型培养基及溶剂兼容。
每次检测大约使用1 x 10-12 mole ATP。用Veritas™微孔板发光光度计(Cat.#
E6501)记录生物发光。 图 A. 测定4种不同培养基 Luria-Bertani
(LB), Mueller Hinton II Broth (MH)， Trypticase Soy
Broth (TSB)或Yeast Peptone Dextrose (YPD) 样品的生物发光。图
B. 测定MH培养基中不含或含有3%溶剂的样品的生物发光。
结 论
新的BacTiter-Glo™微生物活细胞检测是一种用于微生物活细胞检测及定量的简单，灵敏的方法。本检测使用单一试剂从微生物细胞中释放并测量ATP。它是同类型中唯一的均质一步检测。本检测“加入，混合并测量”的方式使其成为抗微生物药物开发及普通微生物学的理想选择。
参考文献
1. DeLuca, M.A. and McElroy, W.D.
(1978) Meth. Enzymol.57, 3-15.
2. McElroy, W.D. and DeLuca, M.A. (1983) J. Applied
Biochem.5,197-209.
3. Lundin, A. and Thore, A. (1975) Anal. Biochem.66,
47-63.
4. Hall, M.P. et al.(1998) Stabilization of firefly
luciferase using directed evolution. In: Bioluminescence
and Chemiluminescence, Perspectives for the 21st Century.
Roda, A.
et al.(eds) New York: John Wiley & Sons., 392-5.
5. Zhang, J-H., Chung, T.D.Y. and Oldenburg, K.F. (1999)
J. Biomol. Screen.4, 67-73.
6. Association of Official Analytical Chemists. (1995)
Bacteriological Analytical Manual, 8th ed. AOAC International,
Gaithersburg, MD.
7. National Committee for Clinical Laboratory Standards.
(2000) Methods for Dilution Antimicrobial Susceptibility
Tests for Bacteria that Grow Aerobically; Approved Standard-Fifth
edition M7-A5. National Committee for Clinical Laboratory
Standards, Wayne, PA. |
|
表1。BacTiter-Glo™ 检测法成功测定的微生物 |
Gram- Bacteria |
Gram+ Bacteria |
Others |
Escherichia coli1 |
Staphylococcus aureus2 |
accharomyces cerevisiae1 |
Psuedomonas aeruginosa2 |
Enterococcus faecalis2 |
Candida albicans2 |
Enterobacter cloacae |
Streptococcus pneumoniae2 |
|
Flavobacterium okeanokoites |
Bacillus subtilis1 |
|
Haemophilus influenzae2 |
Bacillus cereus3 |
|
Proteus vulgaris |
Arthrobacter luteus |
|
Salmonella typhimurium |
|
|
Yersinia enterocolitica3 |
|
|
Francisella philomiragia3 |
|
|
1典型微生物
2药物开发
3生物防御 |
|
|
操作手册
BacTiter-Glo™微生物活细胞检测操作手册#TB337, Promega公司。(http://www.promega.com/tbs/tb337/tb337/html）
定货信息 |
产品 |
包装 |
目录号 |
BacTiter-Glo™ Microbial Cell
Viability Assay |
10ml |
G8230 |
|
10 x 10ml |
G8231 |
|
100ml |
G8232 |
|
10 x100ml |
G8233 |
实验室使用 |
|