Method for detecting the presence of biologically active agents utilizing .sup.13 C-labeled substrates
First Claim
1. A method for detecting the presence of biologically active agents comprising the steps of:
- (a) providing a sterile, sealable container partially filled with a sterile, non-radioactive culture medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2 ;
the remainder of said container being filled with a culture gas;
said container comprising means for introducing a sample of material to be tested for the presence of biologically active agents into the container;
(b) introducing a sample of material to be tested for biological activity into said container and sealing said container;
(c) subjecting the sealed container and its contents to conditions conducive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ;
(d) thereafter measuring the relative proportions of 13 CO2 and 12 CO2 in the culture gas and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas in the container, and(e) comparing the ratio determined in step (d) to a reference standard representing the initial ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas to ascertain whether there has been an increase in the ratio of 13 C16 O2 to 12 CO2 indicative of the presence of a biologically active agent.
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Accused Products
Abstract
A sample of material to be tested for the presence of biologically active agents, such as bacteria, is introduced into a sealable container partially filled with a culture medium comprising a 13 C-labeled fermentable substrate, the remainder of the container being filled with a culture gas, the container and its contents are subjected to conditions conducive to biological activity for a predetermined period sufficient for fermentation of the medium to produce carbon dioxide after which the ratio of 13 CO2 to 12 CO2 in the culture gas in the container is determined and compared to the initial ratio of 13 CO2 to 12 CO2 in the culture gas in order to detect any differences indicating the presence of biologically active agents in the sample.
45 Citations
14 Claims
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1. A method for detecting the presence of biologically active agents comprising the steps of:
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(a) providing a sterile, sealable container partially filled with a sterile, non-radioactive culture medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2 ;
the remainder of said container being filled with a culture gas;
said container comprising means for introducing a sample of material to be tested for the presence of biologically active agents into the container;(b) introducing a sample of material to be tested for biological activity into said container and sealing said container; (c) subjecting the sealed container and its contents to conditions conducive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ; (d) thereafter measuring the relative proportions of 13 CO2 and 12 CO2 in the culture gas and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas in the container, and (e) comparing the ratio determined in step (d) to a reference standard representing the initial ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas to ascertain whether there has been an increase in the ratio of 13 C16 O2 to 12 CO2 indicative of the presence of a biologically active agent. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for detecting the presence of biologically active agents comprising the steps of:
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(a) providing a sterile, sealable container partially fillied with a previously sterilized culture medium to which has been added a sample of material to be tested for the presence of biological activity;
said medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2 ;
the remainder of said container being filled with a culture gas containing a known initial ratio of singly labeled 13 C16 O2 to 12 CO2 ;(b) subjecting the sealed container and its contents to conditions conductive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ; (c) thereafter measuring the relative proportions of 13 CO2 and 12 CO2 in the culture gas and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas in the container; and (d) comparing the ratio determined in (c) to the known initial ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas to ascertain whether there has been an increase in the ratio of singly labeled 13 C16 O2 to 12 CO2 indicative of the presence of a biologically active agent.
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12. A method for detecting the presence of biologically active agents comprising the steps of:
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(a) providing a sterile, sealable container partially filled with a sterile, non-radioactive culture medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2, the remainder of said container being filled with a culture gas;
said container comprising means for introducing a sample of material to be tested for the presence of biologically active agents into the container;(b) introducing a sample of material to be tested fo biological activity into said container and sealing said container; (c) determining the initial ratio of singly labeled 13 C16 O2 to 12 CO2 in said culture gas; (d) subjecting the sealed container and its contents to conditions conductive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ; (e) thereafter measuring the relative proportions of 13 CO2 and 12 CO2 in the culture gas and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas in the container; and (f) comparing the ratio determined in step (e) to the initial ratio determined in step (c) to ascertain whether there has been an increase in the ratio of singly labeled 13 C16 O2 to 12 CO2 indicative of the presence of a biologically active agent.
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13. A method for detecting the presence of biologically active agents comprising the steps of:
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(a) providing a sterile, sealable container partially filled with a sterile, non-radioactive culture medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2, the remainder of said container being filled with a culture gas containing a known initial ratio of singly labeled 13 C16 O2 to 12 CO2 ;
said container comprising means for introducing a sample of material to be tested for the presence of biologically active agents into the container;(b) introducing a sample of material to be tested for biological activity into said container and sealing said container; (c) subjecting the sealed container and its contents to conditions conducive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ; (d) thereafter withdrawing a sample of the culture gas from said container measuring the relative proportions of 13 CO2 and 12 CO2 in said culture gas sample and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in said culture gas sample; and (e) comparing the ratio determined in step (d) to the initial ratio of singly labeled 13 CO2 to 12 CO2 in the culture gas to ascertain whether there has been an increase in the ratio of singly labeled 13 C16 O2 to 12 CO2 indicative of the presence of a biologically active agent.
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14. A method for detecting the presence of biologically active agents comprising the steps of:
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(a) providing a pair of identical, sterile, sealable containers partially filled with equal amounts of a sterile, non-radioactive culture medium comprising a 13 C-labeled substrate fermentable to produce singly labeled 13 C16 O2, the remainder of said container being filled with a culture gas; (b) inoculating one of the containers by introducing a material to be tested for the presence of biological activity into that container and sealing both containers; (c) subjecting the sealed containers and their contents to conditions conducive to biological activity for a predetermined period of time up to 48 hours sufficient for fermentation of the 13 C-labeled substrate to produce singly labeled 13 C16 O2 ; (d) thereafter measuring the relative proportions of 13 C16 O2 and 12 CO2 in the culture gas and determining the ratio of singly labeled 13 C16 O2 to 12 CO2 in the culture gas in each of said containers; and (e) comparing the ratios of singly labeled 13 C16 O2 to 12 CO2 in the inoculated and uninoculated containers to ascertain whether there is a difference in the ratios indicative of the presence of a biologically active agent in the inoculated container.
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Specification