Utilizing lipopolysaccharide in exhaled breath condensate to diagnose gram negative pneumonia

• US 7,828,741 B2
• Filed: 05/23/2005
• Issued: 11/09/2010
• Est. Priority Date: 12/20/2002
• Status: Expired due to Fees

• Current Assignee: Charlotte Mecklenburg Hospital Authority
• Original Assignee: Charlotte Mecklenburg Hospital Authority
• Inventors: Hernandez, Jackeline, Watts, John Albert Jr., Kline, Jeffrey A.
• Primary Examiner(s): Mallari; Patricia C
• Application Number:
  US11/135,265
• Publication Number:
  US 20050208614A1
• Time in Patent Office: 1,996 Days
• Field of Search: 436/900, 600/532
• US Class Current: 600/532
• CPC Class Codes: A61B 5/0813 Measurement of pulmonary parameters by tracers, e.g. radioactive tracers
• CPC Class Codes: A61B 5/083 Measuring rate of metabolism by using breath test, e.g. measuring rate of oxygen consumption
• CPC Class Codes: A61B 5/097 Devices for facilitating collection of breath or for directing breath into or through measuring devices
• CPC Class Codes: A61B 5/412 Detecting or monitoring sepsis
• 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 1/14 Streptococcus; Staphylococcus
• CPC Class Codes: G01N 2400/50 Lipopolysaccharides; LPS
• CPC Class Codes: G01N 33/579 involving limulus lysate
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

A method for determining whether a subject has Gram negative bacterial pneumonia based on the presence of lipopolysaccharide in exhaled breath condensate collected from the subject. The collection devices utilized to collect exhaled breath condensate from both spontaneously breathing and mechanically ventilated subjects and the devices utilized to determine whether lipopolysaccharide is present in the collected exhaled breath condensate.

Claims

1. A method for diagnosing and monitoring intrapulmonary Gram negative bacterial infection in an air-breathing vertebrate subject, comprising:

(a) collecting exhaled breath condensate from an air-breathing vertebrate subject;

(b) providing a reaction chamber with an interior surface, wherein said reaction chamber has a material that binds lipopolysaccharide disposed therein;

(c) delivering at least a portion of the collected breath condensate to the reaction chamber;

(d) washing the reaction chamber with a buffer;

(e) delivering a reaction reagent to the reaction chamber;

(f) determining whether the subject has an intrapulmonary Gram negative bacterial infection based on a physical change that occurs when the at least a portion of the collected exhaled breath condensate is delivered to the reaction chamber, wherein said physical change is caused by the presence of lipopolysaccharide in the exhaled breath condensate, and

(g) identifying the particular strain of the intrapulmonary Gram negative bacteria;

wherein identifying includes identifying the particular strain of intrapulmonary Gram negative bacteria based upon a determination of the O-antigen portion of the lipopolysaccharide molecule,

wherein the binding material is embedded in a matrix surface disposed within the reaction chamber.

2. A method for diagnosing and monitoring intrapulmonary Gram negative bacterial infection in an air-breathing vertebrate subject, comprising:

collecting exhaled breath condensate from the air-breathing vertebrate subject;

providing a reaction chamber, wherein said reaction chamber has a reaction reagent disposed therein;

delivering at least a portion of the collected exhaled breath condensate to the reaction chamber;

determining whether the subject has an intrapulmonary Gram negative bacterial infection based on a physical change that occurs when at least a portion of the collected exhaled breath condensate is delivered to the reaction chamber, wherein said physical change is caused by the presence of lipopolysaccharide in the exhaled breath condensate; and

identifying the particular strain of the intrapulmonary Gram negative bacteria;

wherein identifying includes identifying the particular strain of intrapulmonary Gram negative bacteria based upon a determination of the O-antigen portion of the lipopolysaccharide molecule.

3. A method for diagnosing and monitoring intrapulmonary Gram negative bacterial infection in an air-breathing vertebrate subject, comprising:

collecting exhaled breath condensate from an air-breathing vertebrate subject;

providing a reaction chamber, wherein said reaction chamber has a reaction reagent disposed therein;

delivering at least a portion of the collected exhaled breath condensate to the reaction chamber;

determining whether the subject has an intrapulmonary Gram negative bacterial infection based on a physical change that occurs when the at least a portion of the collected exhaled breath condensate is delivered to the reaction chamber, wherein said physical change is caused by the presence of lipopolysaccharide in the exhaled breath condensate;

visually matching a color change in the reaction chamber to a standard, wherein said standard comprises a printed strip of color patches of increasing hue intensity, and wherein each color patch corresponds to increasing concentrations of lipopolysaccharide, respectively, and

identifying the particular strain of the intrapulmonary Gram negative bacteria;

wherein identifying includes identifying the particular strain of intrapulmonary Gram negative bacteria based upon a determination of the O-antigen portion of the lipopolysaccharide molecule.

4. A method for diagnosing and monitoring intrapulmonary Gram negative bacterial infection in an air-breathing vertebrate subject, comprising:

collecting exhaled breath condensate from an air-breathing vertebrate subject;

providing a reaction chamber with an interior surface, wherein said reaction chamber contains a material that binds lipopolysaccharide disposed therein;

delivering at least a portion of the collected breath condensate to the reaction chamber;

washing the reaction chamber with a buffer;

delivering a reaction reagent to the reaction chamber; and

determining whether the subject has an intrapulmonary Gram negative bacterial infection based on a physical change that occurs when the at least a portion of the collected exhaled breath condensate is delivered to the reaction chamber, wherein said physical change is caused by the presence of lipopolysaccharide in the exhaled breath condensate, and

identifying the particular strain of the intrapulmonary Gram negative bacteria;

wherein identifying includes identifying the particular strain of intrapulmonary Gram negative bacteria based upon a determination of the O-antigen portion of the lipopolysaccharide molecule.