Detection of micro-organisms

• US 8,039,206 B1
• Filed: 08/07/2000
• Issued: 10/18/2011
• Est. Priority Date: 08/06/1999
• Status: Expired due to Fees

• Current Assignee: EMD Millipore Corporation (Merck & Co., Inc.)
• Original Assignee: Millipore Corporation
• Inventors: Keenan, Elizabeth Ann
• Primary Examiner(s): Kim; Taeyoon
• Application Number:
  US10/048,751
• Time in Patent Office: 4,089 Days
• Field of Search: 435/4, 435/7.2, 435/7.31, 435/7.32, 210/500.23, 210/321.6
• US Class Current: 435/4
• CPC Class Codes: B01D 2313/243 Pumps
• CPC Class Codes: B01D 2315/06 Submerged-type; Immersion type
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 63/021 Manufacturing thereof
• CPC Class Codes: B01D 63/024 with a single potted end
• CPC Class Codes: B01D 67/0088 Physical treatment with compounds, e.g. swelling, coating or impregnation
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms

Abstract

Improved methods for detecting microorganisms, such as yeast and bacteria in mixtures, are disclosed. Methods include passing a sample mixture through a filter device, which has been pretreated with a detergent, resuspending the filtrand in the filter membranes and detecting the presence of microorganisms in the filtrand.

Claims

1. A method for recovering microorganisms from a sample mixture, comprising the steps of:

i) passing the sample mixture through a sample inlet that opens into the interior of a filter, the filter comprising a plurality of hollow fibre filter membranes which have been pre-treated with a detergent for increased flow and retention of the microorganisms onto the membranes for re-suspension, the membranes having first and second ends, and an outer side surface and an inner side surface defining a lumen, the first end of each of the membranes being open and communicating with the sample inlet and the second end of each of the membranes being closed such that the flow of the sample mixture is prevented from exiting the second end and directed transverse through the sides of membranes, leaving a filtrand in the lumens of the membranes;

ii) re-suspending the filtrand in the membranes by drawing re-suspension solution into the lumens of the membranes using a syringe;

iii) removing the re-suspended filtrand from the membranes of the filter; and

,

iv) detecting the presence of any microorganisms in the removed filtrand.

2. The method according to claim 1, wherein the re-suspension step further includes passing a solution of lysing agent through the lumen of the membranes.

3. The method according to claim 1, wherein the microorganisms include yeast or bacteria.

4. The method according to claim 1 wherein the membranes have an average pore diameter of 0.2 μ

m.

5. The method according to claim 1 wherein the membranes comprise polypropylene or polysulfone.

6. The method according to claim 1 wherein the detergent comprises a non-ionic detergent.

7. The method according to claim 1 further comprising the step of culturing microorganisms in the filtrand in the filter device for a period of at least 12 hours prior to the detection step.

8. The method according to claim 7, the microorganisms being cultured for at least 24 or 36 hours prior to the detection step.

9. A method for detecting microorganisms in a sample mixture, comprising the steps of:

i) passing the sample mixture through a filter having a sample inlet and an outlet, the sample inlet opening into the interior of the filter, the filter comprising a plurality of hollow fibre filter membranes having an outer side surface and an inner side surface defining a lumen, which have been pre-treated with a detergent for increased flow of the sample and retention of the microorganisms onto the membranes for re-suspension, the sample inlet being open and the outlet being closed such that the flow of the sample mixture is prevented from exiting the outlet and directed transverse through the side surfaces of membranes, leaving a filtrand in the lumens of the membranes;

ii) re-suspending the filtrand in the membranes by drawing lysing agent solution into the lumens of the membranes using a syringe;

iii) removing the re-suspended filtrand from the membranes of the filter; and

,

iv) detecting the presence of cellular ATP in the removed filtrand to determine the presence of any microorganisms.