Sensitive Rapid Detection Method for Viable Bacterial Cells

Author:

MacLeod Robert A.12,Light Marilyn12,White Lloyd A.12,Currie J. F.12

Affiliation:

1. Department of Microbiology, Macdonald College of McGill University, Ottawa, Ontario, Canada

2. Defense Chemical Biological and Radiation Laboratories, Defense Research Board of Canada, Ottawa, Ontario, Canada

Abstract

A rapid sensitive method for the detection of viable bacterial cells is described in which P 32 as inorganic orthophosphate is used to label the cells. Factors affecting the uptake of P 32 by cells as well as the sensitivity of the method have been explored with suspensions of Aerobacter aerogenes . The uptake of P 32 O 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.

Publisher

American Society for Microbiology

Subject

General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine

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