Method for reducing blood carbon dioxide background in bacterial

Details Method for reducing blood carbon dioxide background in bacterial Method for reducing blood carbon dioxide background in bacterial

1989-09-08

1991-02-19

Rosen, Sam

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving viable micro-organism

435 31, 435 34, C12Q 102

Patent

active

049943782

ABSTRACT:
A non-ionic lytic agent, preferably saponin for reducing the background carbon dioxide produced by blood cell metabolism is used in the testing of cultures for the presence of microorganisms. The hemolytic agent saponin is combined with a phospholipid, preferably L-.alpha.-Lecithin (phosphatidylcholine), to form mixed micelles which protect saponin from the effects of heat sterilization and high blood cholesterol levels, thus maintaining the lytic activity of saponin. The phospholipid/saponin mixed micelles are added to non-radiometric culture media vials such as Bactec.RTM. NR6A, NR7A and NR8A. The media vials are used in the Bactec.RTM. NR-660 and NR-730 instruments. However, the present invention may also be used in radiometric media such as Bactec.RTM. culture vials 6, 7 and 8 for reducing background carbon dioxide levels detected in C.sup.14 radiometric instruments such as the Bactec.RTM.-460, or in other instruments designed to detect bacteria in the presence of mammalian cells by monitoring the increase in CO.sub.2 produced by the metabolizing bacteria.

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Interaction of Saponin and Digitonin with Black Lipid Membranes and Lipid Monolayers, Gogelein et al., Biochimica et Biophysica Acta, 773, 32-38 (1984).
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The Phospholipid-Cholesterol Interaction Kinetics of Water Permeability in Liposomes, Bittmann et al., Biochemistry, 11, 4831-4839, (1972).
Fusion of Dipalmitoylphosphatidylcholine Vesicles, Schullery et al., Biochemistry, 19, 3919-3923 (1980).

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