Method for the detection of biologically active agents

• US 4,971,900 A
• Filed: 03/21/1988
• Issued: 11/20/1990
• Est. Priority Date: 04/06/1984
• Status: Expired due to Fees

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Perks, H. Mark, Tice, Gregory, Sussman, Mark L., Ahnell, Joseph E.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Waack, J. D.
• Application Number:
  US07/171,499
• Time in Patent Office: 974 Days
• Field of Search: 435/29, 435/34, 435/35, 435/38, 435/39, 435/40, 435/291, 435/299, 435/313, 435/807, 422/68, 436/146
• US Class Current: 435/29
• CPC Class Codes: C12M 41/34 of gas
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 2304/46 Carbon dioxide
• CPC Class Codes: Y10S 435/807 Gas detection apparatus
• CPC Class Codes: Y10T 436/235 In an aqueous solution [e.g., TOC, etc.]

Abstract

An apparatus and method for the detection of the growth of microorganisms through infrared analysis of a sample of the gas produced by growth of the microorganism is descirbed. In the method, a sample of the headspace gas in a vial containing a growth medium which has been inoculated with a sample suspected of containing a microorganism is removed and transferred to a sample cell where infrared analysis is used to determine the presence of carbon dioxide, if any, produced by the growth of the microorganism.

Claims

1. A method for detecting the presence of microorganisms in a sample of material to be analyzed comprising the steps of:

(a) introducing a test sample into a sterile vial having culture medium and head space gas wherein the culture medium has a carbon source and the head space gas has a known initial CO₂ concentration;

(b) incubating the vial at conditions conducive to metabolism of microorganisms;

(c) introducing, into a test head for determining carbon dioxide content of gas therein, a culture gas having a CO₂ concentration substantially equal to the known initial CO₂ concentration of the head space gas;

(d) determining the CO₂ concentration level of the culture gas with the test head, the test head having sensing and motive components for use in sampling the head space gas in the vial;

(e) establishing a fluid communication path between the head space and the test head;

(f) mixing the culture gas in the test head with the head space gas;

(g) measuring in the test head the concentration of CO₂ in the mixture of culture gas and the head space gas; and

(h) comparing the measured CO₂ concentration of the mixture with the CO₂ concentration of the culture gas determined in step (d) to detect the presence of microorganisms.

2. The method of claim 1, further characterized by the level of CO₂ concentration of a mixture of the culture gas with the test head space gas before said measuring step being within the range of between about 1 and 10 percent.

3. The method of claim 2, further characterized by said level of CO₂ concentration of the mixture being within the range of between about 2 and 5 percent.

4. The method of claim 1, wherein the sensing components of the test head have a measuring cell and said step of establishing the fluid communication path enables the measuring cell to measure the CO₂ concentration in the mixture.

5. The method of claim 4, further characterized by flushing the fluid communication path with the culture gas before step (d).

6. The method of claim 5, further characterized by providing the culture gas with a level of CO₂ concentration within the range of between about 1 and 10 percent.

7. The method of claim 1, further characterized by using whole blood as the sample to be tested.

8. The method of claim 1, further characterized by analyzing a plurality of culture vials in sequence.