Method and unit for preparing a sample for the microbiological analysis of a liquid

• US 9,410,181 B2
• Filed: 05/23/2007
• Issued: 08/09/2016
• Est. Priority Date: 06/27/2006
• Status: Active Grant

• Current Assignee: EMD Millipore Corporation (Merck & Co., Inc.)
• Original Assignee: EMD Millipore Corporation (Merck & Co., Inc.)
• Inventors: Ribault, Sebastien, Waiche, Gael
• Primary Examiner(s): Gebreyesus, Kagnew H
• Assistant Examiner(s): MARTIN, PAUL C
• Application Number:
  US11/805,539
• Publication Number:
  US 20070298451A1
• Time in Patent Office: 3,366 Days
• Field of Search: 435/4, 435/30, 435/288.1
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/24 Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
• CPC Class Codes: G01N 2001/4088 filtration

Abstract

The preparation unit comprises a body (2) within which are fixed a first (3) and a second (4) membrane, said first membrane (3) having a first predetermined pore diameter and said second membrane (4) a second predetermined pore diameter smaller than said first predetermined pore diameter of said first membrane (3), said body also comprising means (16, 20) for retrieving said sample collected on said second membrane (4).

The method of preparing such a sample comprises the step of procuring such a preparation unit (1), the step of passing a predetermined volume of said liquid through said first membrane and through said second membrane (4) in order to collect a sample on said second membrane (4); and the step of retrieving said sample so collected on said second membrane (4).

Claims

1. A method of preparing a sample for the microbiological analysis of a liquid containing microorganisms of several different types each having predetermined morphological characteristics, which method comprises the step of determining the size of undesired microorganisms in said sample;

the step of procuring a unit for preparing said sample comprising a generally tubular body within which are fixed a first and a second membrane, said body having an opening for introduction of said liquid, a first compartment between said introduction opening and said first membrane, a second compartment between said first and second membranes and a third compartment on the side of said second membrane opposite the side facing said first membrane,

the step of selecting said first membrane in response to the determined size of the undesired microorganisms, to have a first pore diameter small enough to retain said undesired microorganisms and large enough to allow desired microorganisms to pass through,

the step of selecting said second membrane to have a predetermined second pore diameter less than said predetermined first pore diameter of said first membrane and small enough to retain said desired microorganisms;

the step of passing a predetermined volume of said liquid from the first to the second compartment through said first membrane;

the step of passing all the filtrate, having so reached the second compartment, from the second to the third compartment through said second membrane in order to collect said sample on said second membrane with said desired microorganisms selected from said different types of microorganisms which are those of which the size is less than the predetermined pore diameter of said first membrane and greater than the predetermined pore diameter of said second membrane;

the step of retrieving said sample so collected on said second membrane for further analysis of the desired microorganisms contained in said sample;

wherein the steps of passing said liquid through said membranes are performed by centrifuging said preparation unit.

2. A method according to claim 1, wherein the step of procuring a preparation unit comprises the step of selecting a preparation unit of which the body comprises a first portion to which is fixed said first membrane and a second portion to which is fixed said second membrane, said portions being detachable from each other, and in that the step of retrieving said sample comprises the step of detaching said second portion of said body from said first portion of said body.

3. A method according to any one of claims 1 to 2, wherein the step of procuring a preparation unit comprises the step of selecting as said first membrane a membrane of pore diameter greater than 1 μ

m.

4. A method according to any one of claims 1 to 2, wherein the step of procuring a preparation unit comprises the step of selecting as said second membrane a membrane of pore diameter less than 0.65 μ

m.

5. A method according to claim 4, wherein the step of procuring a preparation unit comprises the step of selecting as said second membrane a membrane of pore diameter less than 0.1 μ

m.

6. The method of claim 1, wherein said further analysis comprises extracting said second membrane and placing it in contact with a gel growth medium, followed by incubating said microorganisms on said second membrane.

7. The method of claim 1, wherein said further analysis comprises treating said second membrane with a reagent capable of revealing the presence of ATP of the microorganisms on said second membrane.

8. The method of claim 1, wherein said further analysis comprises analyzing the DNA of the microorganisms on said second membrane.