Methods for diagnosing infectous diseases and methods for detecting and identifying microorganisms

• US 5,358,846 A
• Filed: 02/02/1990
• Issued: 10/25/1994
• Est. Priority Date: 04/20/1988
• Status: Expired due to Term

• Current Assignee: FUSO Yakuhin Kogyo Kabushiki Kaisha
• Original Assignee: FUSO Yakuhin Kogyo Kabushiki Kaisha
• Inventors: Matsuhisa, Akio, Ohno, Tsuneya
• Primary Examiner(s): Parr, Margaret
• Assistant Examiner(s): Sisson, Bradley L.
• Application Number:
  US07/449,931
• Time in Patent Office: 1,726 Days
• Field of Search: 435/6, 435/7.1, 435/7.31, 435/7.32, 435/7.5, 435/822, 435/911, 435/973, 536/24.32, 935/76-78
• US Class Current: 435/6.15
• 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 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/6888 for detection or identification of organisms
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: Y10S 435/822 using bacteria or actinomycetales
• CPC Class Codes: Y10S 435/911 using fungi
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte

Abstract

The invention provides methods and materials for the detection and identification of biotin-containing bacteria and/or yeast in phagocyte-containing samples obtained from patients.

Claims

1. In a method for detecting biotin-containing bacteria or yeast in a sample obtained from a patient comprising fixing a phagocyte containing specimen obtained from the patient on a solid support for a time and under conditions sufficient for said phagocytes to become fixed to said support, the improvement comprising exposing the fixed phagocytes to a reagent having a preferential binding affinity for biotin, and detecting any product formed by binding of the reagent to bacterial or yeast biotin contained within said fixed phagocytes.

2. The method of claim 1 wherein said reagent is selected from the group consisting of avidin or streptavidin.

3. The method of claim 1 wherein the step of detecting said product comprises exposing said product to biotin labelled with a detectable label and detecting any complex formed between the product and the labelled biotin.

4. The method of claim 1 wherein said detectable label is an enzyme, an antibody, a dye, gold, or a radioisotopic element.

5. The method of claim 1 wherein the phagocyte-containing specimen is selected from the group consisting of blood, ascites fluid and pus.

6. The method of claim 1 wherein said specimen is fixed to said solid support by a fixing step which comprises immersing the specimen in Carnoy'"'"'s B fixative.

7. The method of claim 1 further comprising identification of the specific bacteria or yeast causative agent of an infection detected by means of the formation of said product through the subsequent step of hybridization of bacteria or yeast nucleic acids in said fixed phagocytes with one or more detectable oligonucleotide probes specific for bacterial or yeast species.

8. The method of claim 7 wherein said detectable oligonucleotide probe or probes are labelled with an enzyme, an antibody, gold, or a radioisotopic element.