MXPA98002029A - Determination of microorganisms not parafinofili - Google Patents

Abstract

A method for determining the presence or absence of a non-parafinophilic microorganism in a sample taken from a patient includes providing a container containing an aqueous solution and inoculating the aqueous solution with the sample. A slide coated with a carbon source is placed inside the container. By analyzing the slide after exposure of the sample, the presence or absence of a non-parafinophilic microorganism can be determined in the sample. An associated apparatus is also described

Description

DETERMINATION OF NON-PARAFINOPHILIC MICROORGANISMS BACKGROUND OF THE INVENTION This invention is to a method for determining the presence or absence of a non-parafinophilic microorganism in a sample and associated apparatus. The identification of non-parafinophilic microorganisms in a clinical sample is an important part of the medical treatment of patients. Often, instructed assumptions are made about the nature of the microorganisms involved. It would therefore be beneficial to improve the identification process of those microorganisms with a simple and effective method and apparatus. As used herein, the term "non-paraffinic microorganism" represents any microorganism supported by a carbon source other than paraffin Examples of such nonparaffinophilic microorganisms include, but are not limited to the following: Mycobacterium tuberculosis. Mycobactenum paratuberculosis; Mycobacterium leprae; Staphylococcus Streptococcus, E. Coli, Listeria; Brucellae, Humemophilus Treponema; Pneumococcus, Clostridium, Cryptococcus, Coccidioides and Histoplasma Likewise, as used herein, the term "patient" refers to a member of the animal rowing including human beings, whose body sample is being processed by the method and apparatus of the animal. Invention U.S. Patent Nos. 5,153,119 and 5,316,918 disclose methods and apparatus for identifying and testing the antibiotic sensitivity of Mycobacterium avium-intracellulare ("MAI"), a nonparaffinophilic microorganism. The inventor named in those patents is Robert A. Ollar, one of the co-inventors of the invention described herein. This method involves providing a container containing an aqueous solution and injecting a sample into the solution. After this, a slide coated with paraffin is placed inside the container. The slide is then observed for the presence or absence of growth of MAL Despite the efficient, effective and economical method described in Dr. Ollar's patents, there is still a need for a simple and effective method to determine the presence or absence of a non parafinophilic microorganism.

BRIEF DESCRIPTION OF THE INVENTION The invention has met or exceeded the needs mentioned above as well as others. A method for determining the presence or absence of a non-parafinophilic microorganism in a sample taken from a patient comprising providing a container containing an aqueous solution and inoculating the aqueous solution with the sample. A slide coated with a carbon source is placed inside the container. By analyzing the slide after exposure to the sample, the presence or absence of a non-parafinophilic microorganism can be determined in the sample. An associated apparatus is also described. The apparatus comprises a container for holding an aqueous solution and a slide coated with a carbon source adapted to be placed in the container. The carbon source bathes the nonparaffinophilic microorganism. The carbon source can be included in a gelatinous matrix that is attached to the slide or can be an ionically bound or affinity carbon source linked to a plurality of gel beds that are adhered to the slide.

BRIEF DESCRIPTION OF THE DRAWING A complete understanding of the invention can be achieved from the following detailed description of the invention when read in conjunction with the accompanying drawing which shows a front elevation view of a test tube holding a slide covered with a fountain. of carbon in an aqueous solution inoculated with a sample.

DETAILED DESCRIPTION The method and apparatus of the invention provide an efficient, effective and economical way of identifying a nonparaffinophilic microorganism. Referring now to the sole Figure, a nonparaffinophilic microorganism identification apparatus 10 is shown. The apparatus 10 includes a standard test tube 12 which contains an aqueous solution 13 (such as Czapek broth) and a cotton plug 16 for sealing the test tube 12. In accordance with the invention, a sample to be tested for the presence or absence of a non-paraffinophilic microorganism is inoculated into the aqueous solution 13. A slide 18 having a coating comprising or containing a carbon source 20 is then placed inside the tube. Test 12. It will be appreciated that the aqueous solution 13 should not contain any carbon source, as desired to provide a single carbon source 20 on the slide 18 in order to effectively culture the non-parafinophilic microorganism to be identified in the slide 18 and not in the aqueous solution 13. The culture on the slide 18, which may or may not be observable by the human eye without help, may be analyzed to determine the presence or absence of a non-parafinophilic microorganism. Preferably, a minimum incubation time of twenty-four (24) hours is necessary for growth to occur. To analyze the slide 18 after the incubation period, the slide 18 can be scraped using a spatula sterilized to the flame and subcultured on a tryptic soy agar (TSA). Whether scrapings include growth on TSA can be analyzed using classical microbiological procedures or can be analyzed using an extraction process that involves either extraction of organic solvent or column chromatographic extraction. The sample to be inoculated into test tube 12 can be a blood sample; any biopsy or tissue sample; stomach fluid; urine; cerebrospinal fluid; Nasopharyngeal mucosa or saliva. These samples can be obtained from the patient in the doctor's office or in the emergency room of a hospital, for example, by means of techniques known in normal standard media. The carbon source 20 on the slide 18 may include a gelatinous matrix containing a carbon source. A carbon source may be one or more of those selected from the group comprising glucose, fructose, glycornol, mannitol. asparagine and casein among others. Another embodiment may include providing a coating and a slide with an adhesive and securing a plurality of gel beds to the adhesive. The carbon source can be either linked ionically or by affinity to the gel beds. The slide 18 with the gelatinous matrix containing a carbon source can be prepared by the following method. A container, such as a laboratory beaker, is first filled with 100 ml of distilled water. Two (2) grams of agar (gelatinous matrix) and three (3) grams of carbon source (such as glucose) are placed into the laboratory ford. This mixture is then boiled and the sterilized steam and molten gelatinous matrix is poured into the petri dish, which is seated on a hot plate. In this way the gelatinous matrix / carbon source remains fused. After this, a sterile slide 18 is dropped into the gelatinous matrix / carbon source and becomes coated therewith. The coated slide is removed from the petri dish and allowed to stand for a minute or two to solidify the coating 20 therein. The slide with the coating of a gelatinous matrix containing a carbon source is then easily placed in the test tube 12 containing the aqueous solution 13 and the sample. An alternative method of preparation of the slide involves first coating the slide with an adhesive, such as collodion and then applying a plurality of gel beds (commercially available from Pharmacia of Parsippany, New Jersey) to the adhesive. The gel beds are approximately one meter in diameter. The slide containing the coating of the gel beds is immersed in a pH regulating agent containing the carbon source (such as glucose) to bind the carbon source to the gel beds either ionically or by affinity. Non-paraffinophilic microorganisms that can be identified using the method of the invention include any microorganism supported by a carbon source other than paraffin. Non-paraffinophilic microorganisms include, but are not limited to, Mycobacterium tuberculosis; Mycobacterium paratuberculosis; Mycobacterium leprae; Staphylococcus: Streptococcus; E. Coli; Listeria; Brucellae; Humemophilus; Treponema; Pneumococcus; Clostridium; Cryptococcus; Coccidioides: and Histoplasma. It will be appreciated that a method for determining the presence or absence of a nonparaffinophilic microorganism in a sample of a non-parafinophilic microorganism in a sample and associated apparatus has been described. The method is effective and efficient and does not involve the use of expensive and complicated equipment. An associated apparatus is also described. While the specific embodiments of the invention have been described, those skilled in the art will appreciate that various modifications and alterations to those details could be developed in light of the general teachings of the description. Accordingly, the particular provisions described are intended to be illustrative only and do not limit the scope of the invention to be given by the broad scope of the appended claims and any and all equivalents thereof.

Claims (1)

1. CLAIMS 1 A method for determining the presence or absence of a non-parafinophilic microorganism in a sample taken from a patient, the method comprising providing a container containing an aqueous solution, inoculating the aqueous solution with the sample, placing a slide within said container coated with a carbon source, and analyzing the slide after exposure to said sample to determine the presence or absence of the non-parafinophilic microorganism The method of claim 1, which includes employing as said slide a one coated with a gelatinous matrix containing A carbon source 3 The method of claim 1, which includes providing the slide by first adhering a plurality of gel beds to the slide and then bonding the carbon source 4 to the gel beds. The method of claim 3, wherein the carbon source is ionically bound to the gel beds 5 The method of the claim 3 wherein the carbon source is affinity bound to the gel beds 6 The method of claim 3, which includes adhering the gel beds to the slide by means of an adhesive 7. The method of claim 6, which includes employing collodion as said adhesive. The method of claim 1, which includes employing as the carbon source one or more of the group comprising glucose, fructose, glycerol, mannitol, asparagine and casein. The method of claim 1, which includes using as sample one selected from the group comprising blood, stomach fluid, urine, cerebrospinal fluid, nasopharyngeal mucosa and saliva. 10. An apparatus for facilitating the determination of the presence or absence of a nonparaffinophilic microorganism in a sample taken from a patient, the apparatus comprising: a container for holding an aqueous solution; and a slide coated with a carbon source, the slide adapted to be placed in said container. The apparatus of claim 10, wherein the slide is coated with a gelatinous matrix containing the carbon source. 12. The apparatus of claim 10, wherein the slide is coated with a plurality of gel beds that have attached thereto the carbon source. The apparatus of claim 12, wherein the carbon source is ionically bound to the gel beds. The apparatus of claim 12, wherein the carbon source is affinity bound to the gel beds 15. The apparatus of claim 12, wherein the gel beds are adhered to the slide by an adhesive. 16. The apparatus of claim 15, wherein the adhesive is collodion. The apparatus of claim 10, wherein the carbon source is one or more of the group consisting of glucose, fructose, glycerol, mannitol, asparagine and casein.