Macleod R A, Light M, White L A, Currie J F
Department of Microbiology, Macdonald College of McGill University, Ottawa, Ontario, Canada.
Appl Microbiol. 1966 Nov;14(6):979-84. doi: 10.1128/am.14.6.979-984.1966.
A rapid sensitive method for the detection of viable bacterial cells is described in which P as inorganic orthophosphate is used to label the cells. Factors affecting the uptake of P by cells as well as the sensitivity of the method have been explored with suspensions of Aerobacter aerogenes. The uptake of PO(4) is dependent on several factors. Of various incubation media tested, one composed of 0.005 m KCl, 0.002 m MgSO(4) and 10 mg/ml of glucose was found to best stimulate the uptake of the tracer. Incubation time and temperature and level of isotope and of unlabeled P also affected uptake. Labeled cells were collected on a membrane filter for measurement of radioactivity. Under optimal conditions, as few as 23 viable cells per milliliter were detected in 1 hr with 95% confidence.
本文描述了一种快速灵敏的检测活细菌细胞的方法，该方法使用无机正磷酸盐形式的磷来标记细胞。利用产气气杆菌悬浮液，研究了影响细胞对磷摄取的因素以及该方法的灵敏度。磷酸根的摄取取决于几个因素。在测试的各种培养基中，发现一种由0.005m氯化钾、0.002m硫酸镁和10mg/ml葡萄糖组成的培养基能最佳地刺激示踪剂的摄取。孵育时间、温度、同位素和未标记磷的水平也会影响摄取。将标记的细胞收集在膜滤器上以测量放射性。在最佳条件下，每毫升低至23个活细胞在1小时内即可被检测到，置信度为95%。
