WO2009079232A2

WO2009079232A2 - Method and apparatus for micro-organism capture

Info

Publication number: WO2009079232A2
Application number: PCT/US2008/085659
Authority: WO
Inventors: Paul W. Fogle, Jr.; Jason Kenneth Holt; Brian G. Shelton
Original assignee: Pathogen Control Associates (Dba Pathcon Laboratories); Elutrasep, Inc.
Priority date: 2007-12-05
Filing date: 2008-12-05
Publication date: 2009-06-25
Also published as: WO2009079232A9; WO2009079232A3

Abstract

A micro-organism capture apparatus, including a housing having an inlet and outlet, the housing including therein a filter having an attracting mechanism for attracting and retaining micro-organisms, and a release mechanism for releasing the attracting mechanism operatively attached to the housing. A kit for sampling a liquid for micro-organisms. A method of attracting and retaining micro-organisms by flowing liquid through the micro-organism capture apparatus, and attracting, retaining, and concentrating micro-organisms within the liquid with the attracting mechanism. A method of isolating micro-organisms from a liquid sample by flowing liquid through the micro-organism capture apparatus, attracting, retaining, and concentrating micro-organisms within the liquid with the attracting mechanism, releasing the attracting means with the release mechanism, and isolating the micro-organisms retained by eluting the micro-organisms from the attracting mechanism. A method of indicating a liquid is in need of remediation.

Description

METHOD AND APPARATUS FOR MICRO-ORGANISM CAPTURE

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority 35 U. S. C. Section 119(e) of United States Provisional Patent Application No. 60/992,509, filed December 5, 2007, which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

(1) Field of the invention

[0002] The present invention relates to capture by filtration. In particular, the present invention relates to filtration as a part of a system for capturing micro-organisms such as viruses by using an attraction mechanism.

(2) Description of the Prior Art

[0003] There are many viruses in our environment that can contaminate drinking water for human consumption. For example, human Norovirus, a genus of the family Caliciviridae, constitutes up to 50% of the reported cases of gastroenteritis. Persons with Norovirus are generally characterized by the symptoms of nausea, vomiting, diarrhea, and abdominal pain. The Norwalk virus is a common type of Norovirus. Genotypes I, II, and IV of Norovirus typically infect humans. Infection generally occurs through contaminated food and drinks (i.e. drinking water). Livestock waste has been shown to test positive for Norwalk-like viruses, other Noroviruses, and other types of caliciviruses.

[0004] Currently, there are no user-friendly tests to sample large volumes of water to test for these environmental contaminants. Elution volumes and concentrated retainate of viruses from current methods of testing are still too large for manageable analysis. [0005] The top two current methodologies are the Cuno, 1 MDS charged fiberglass filter and hollow-fiber ultrafiltration to concentrate viruses from environmental water sources, wastewater effluent, and finished, potable drinking water. These methods have several drawbacks. For example, large volumes of water (100 L - 1000L) are used to concentrate and detect viruses, which is often not practical. The methodologies may take days to weeks to concentrate the viruses and complete the analysis. Typically less than 10% of the concentrated sample is analyzed due to high volume of concentrated retainate. For current methodologies, recovery is typically less than 60% for most viruses. The effective sample volume from 100 L of water is typically less than 6 L. Furthermore, these procedures are costly, running around $1 ,000 per sample. [0006] Other methodologies and filters are able to capture viruses and concentrate them but are not able to elute the viruses for detection or they have low recoveries as described below. Also, most methodologies use antibody coated microspheres or particles which will only capture a specific target virus and not all of the viruses present in the sample population.

[0007] International Patent Publication No. WO/2001/0011006 to Fluid Technologies, PLC discloses a method of detecting yeast in beer by using a hollow fiber membrane filter to collect a sample and then analyzing the sample for the presence of yeast. [0008] United States Patent No. 6,051 ,189 to Wick, et al. discloses a system and method for detection, identification, and monitoring of submicron sized particles. The method generally involves collecting a sample, extracting existing submicron particles from the collected sample based on density, purifying the extracted submicron particles by concentrating the extracted submicron particles based on size, and, detecting and identifying the purified extracted submicron particles based on size and density thereby determining submicron particles present in the collected sample. The submicron particles detected and identified include viruses and virus-like agents such as prions. Essentially, the method relies on the synergy of centrifugation and ultrafiltration. [0009] United States Patent No. 5,498,550 to Fujiwara, et al. discloses a method and device for collecting a specimen and for preparing a specimen characterized by magnetically labeling a specimen with a magnetic-labeled body composed of a micro- particle of a magnetic substance and an antibody or antigen bound thereto, and applying a gradient magnetic field to the magnetic-labeled specimen to locally concentrate the labeled specimen to a predetermined position and recovering an immunocomplex efficiently. Through this method, a small amount of a virus can be detected. [00010] United States Patent No. 3,970,518 to Giaever also relates to magnetic separation and discloses a method for sorting out and separating a select cell population from a mixed cell population comprising the steps of applying to the surface of small magnetic particles a coating of an antibody to the select cell, bacteria or virus population: moving these antibody-coated magnetic particles through a liquid containing the mixed population whereby the members of the cell, bacteria or virus population become affixed to the antibody coatings on the particles, separating the coated magnetic particles with such members attached thereto from the rest of the mixed population; introducing the coated magnetic particles and attached members into a solution of cleaving agent whereby the bonds between the antibody and the members of the select population is broken and separating the antibody-coated magnetic particles and solution containing the select population of cells, bacteria or viruses from each other.

[00011] United States Patent No. 4,663,277 to Wang discloses a method for detection of viruses in a specimen, wherein the specimen, treated to remove undesired components, is contacted with an extended solid phase having conjugated thereon antiviral antibody (Ab. sub. v) to form immuno-complexes with antigens characteristic of the viruses to be detected; the extended solid phase is separated from the specimen; the separated extended solid phase is contacted with a mobile solid phase consisting of dispersed microspheres having conjugated thereto said Ab. sub. v to bind the microspheres to the immuno-complexes; the extended solid phase is separated from the mobile solid phase; and the presence of microspheres bound to the extended solid phase is detected, whereby the presence of viruses in said specimen is detected or determined.

[00012] There is an Environmental Protection Agency (EPA) initiative to develop a regulation to monitor finished drinking water for viruses by 2009, but currently, there are no regulations to address the presence of viruses due to the limitations described above. Currently, no feasible method exists for the testing of viruses in drinking water. Therefore, there is a need for a method of testing for viruses in drinking water that is quick, manageable, and inexpensive. Furthermore, there is a need for a rapid pre- analytical method to concentrate viruses and other cellular material for rapid detection with greater efficiency and from lower sample volumes than currently available.

SUMMARY OF THE INVENTION

[00013] The present invention provides for a micro-organism capture apparatus, including a housing having an inlet and outlet, the housing including therein a filter having an attracting mechanism for attracting and retaining micro-organisms, preferably viruses, and a release mechanism for releasing the attracting mechanism operatively attached to the housing.

[00014] The present invention provides for a kit for sampling a liquid for microorganisms, including the micro-organism capture apparatus, tubing, luer hose barb fittings, luer lock end caps, a plug, mailing supplies, and an instruction manual. [00015] The present invention also provides for a method of attracting and retaining micro-organisms, including the steps of flowing liquid through the micro-organism capture apparatus, and attracting, retaining, and concentrating micro-organisms within the liquid with the attracting mechanism.

[00016] The present invention further provides for a method of isolating microorganisms from a liquid sample, including the steps of flowing liquid through the microorganism capture apparatus, attracting, retaining, and concentrating micro-organisms within the liquid with the attracting mechanism, releasing the attracting means with the release mechanism, and isolating the micro-organisms retained by eluting the microorganisms from the attracting mechanism.

[00017] The present invention also provides for a method of indicating a liquid is in need of remediation, including the steps of isolating at least one micro-organism with the micro-organism capture apparatus, and positively detecting the micro-organism with an assay, and indicating that the liquid is in need of remediation.

BRIEF DESCRIPTION ON THE DRAWINGS

[00018] Other advantages of the present invention will be readily appreciated as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings, wherein: [00019] FIGURE 1A is a representative cross-sectional view of a hollow-fiber filter described in the present invention, shown in dead-end mode and FIGURE 1 B is a close-up view of the filter;

[00020] FIGURE 2 is a representation of a flat-bed membrane filter described in the present invention;

[00021] FIGURE 3 is a illustration of the micro-organism capture apparatus with a hollow-fiber membrane; and

[00022] FIGURE 4A is an illustration of the flow of liquid in the micro-organism capture apparatus and FIGURE 4B is an illustration of the collection of a sample with the micro-organism capture apparatus.

DETAILED DESCRIPTION

[00023] The present invention provides a rapid and sensitive assay for the capture of micro-organisms, preferably viruses, from the environment, particularly from drinking water.

[00024] The term "micro-organism" as used herein includes viruses as well as bacteria, protozoa, and fungi, various types of which are further described below. Bacteria, protozoa, and fungi are relatively easy to capture by size exclusion with the filter of the present invention. Viruses pose the most difficult challenge in testing liquids (ex. potable water) because size-exclusion filtration has either not been effective in retaining viruses due to their small size, or the virus becomes trapped in the membrane and is extremely difficult to elute from the filter,, thus having a very low, or inconsistent recovery rate. Therefore, the present invention uses a novel attracting mechanism to attract and retain viruses as well as other micro-organisms. The present invention is efficient in capturing each of these types of micro-organisms, especially viruses. [00025] More specifically, the present invention provides a micro-organism capture apparatus 10 including a housing 12 having an inlet 14 and an outlet 16, shown generally in FIGURES 1-3. The housing 12 includes therein a filter 18 including an attracting mechanism 20 for attracting and retaining micro-organisms, which is preferably a positively charged nano-particle. The housing 12 also includes a release mechanism 22 for releasing the attracting mechanism 20 for concentration and elution of the micro-organisms.

[00026] The housing 12 can be made of any suitable materials such as a plastic. The inlet 14 is designed to receive a liquid from a source such as a tube 30 through which the liquid is flowing. The liquid can be received from a smaller sample 32 than the liquid source (as shown in FIGURE 4B), or directly from the liquid source itself. The tube 30 can include a pump 34, such as a peristaltic pump, to extract the liquid from the sample 32 or liquid source. The outlet 16 dispenses the liquid that has flowed through the attracting mechanism 20 (i.e. the filtrate) to another tube or collection vessel 36 (shown in FIGURE 4A). The housing is manufactured according to methods known in the art. [00027] The filter 18 can be any suitable filter that is able to retain the attracting mechanism 20 within the filter 18 while allowing the micro-organism to collide with the attracting mechanism 20 when a liquid is flowed through the filter 18. In other words, the pore size 24 of the filter 18 is less than the diameter of the attracting mechanism 20 but large enough to allow for flow of the liquid readily through the filter 18. Thus, the filtration of the micro-organism attracting mechanism is size-exclusion filtration. Preferably, the filter 18 is a hollow-fiber filter membrane designed for microfiltration, as shown in FIGURES 1A and 1 B; however, any other filter 18 or material with the necessary pore size can be used, such as a micro-porous flat-bed membrane shown in FIGURE 2. The filter 18 of FIGURE 1 can also be used with a pressure regulator valve attached to a loop to create a restricted flow, without dead-end filtration, but creating enough pressure to enable titration across the hollow-fiber membrane and creating a concentrated sample retainate containing the attracting mechanism particles. The filter 18 can be manufactured by methods known in the art, and particularly, the attracting mechanism 20 can be pre-loaded within the filter 18.

[00028] The attracting mechanism 20 is preferably a positively charged material or structurally modified magnetic particle, such as magnetic beads, that is designed to attract and retain viruses by electrostatic interaction in a liquid sample. The attracting mechanism 20 can also be negatively charged in order to attract positively charged micro-organisms. Preferably, the attracting mechanism 20 is in the shape of nano- spheres or nano-particles. Although nano-particles provide a high liquid-to-surface contact area, one skilled in the art can easily understand that one can use an attracting mechanism 20 other than nano-particles, i.e. charged particles of various shapes and sizes, and with various modified surfaces to attract targeted viruses, protozoa and other micro-organisms. The attracting mechanism 20 can include an antibody coating (with a positive or negative charge, such as silica coated Ab beads) or any other coating with which to attract and retain the viruses or other micro-organisms. In other words, the charged attracting mechanism 20 attracts any oppositely charged body that it comes in contact or close proximity with, i.e. viruses. Preferably, the attracting mechanism 20 can attract and retain several different viruses. Alternatively, the attracting mechanism 20 can be designed to attract and retain a specific micro-organism of interest. Thus, viruses and other micro-organisms that would normally pass through filtration membranes are retained by the attracting mechanism 20.

[00029] The filter 18 with the attracting mechanism 20 acts as a fluidized bed for collecting and concentrating viruses and micro-organisms therein. Retention of the attracting mechanism 20 can be accomplished by establishing a fluidized bed in the filter 18. Examples include dead-end filtration, pressurization (when the filter is a hollow-fiber membrane), or other filtration methods that use a pore size small enough to retain the attracting mechanism 20 in the contact zone, and also provide for easy and efficient elution of the attracting mechanism 20.

[00030] The filter 18 includes operatively attached thereto a release mechanism 22. The release mechanism 22 is necessary for releasing the attracting mechanism 20 from the filter 18. Concentration and elution of the viruses and micro-organisms can then be performed on the attracting mechanism 20. When the filter 18 is in the form of a hollow- fiber membrane, shown in FIGURE 1 , the release mechanism 22 can be a removable cap, valve, seal, plug-end, or any form of restriction that will create a backpressure to the fluidized bed, causing the fluid to permeate the pores 16 of the filter 18. In the case of other filtration, such as flat-bed filtration (FIGURE 2), the release mechanism 22 can be the removal of a disposable flat-bed filter 18 containing the attracting mechanism 20. [00031] Preferably, the micro-organisms that are attracted and retained with the attracting mechanism 20 include Noroviruses, Adenoviruses and polioviruses. Also, other waterborne micro-organisms such as Cryptosporidium parvum, Giardia lamblia and bacteria such as Bacillius anthraicis and Vibrio cholera can be attracted with the embodiment of an antibody-coated bead. Any other micro-organisms that are potentially harmful to humans and can be present in drinking water or other liquids, or are contaminants or indicators of contamination can be attracted with the device of the present invention as well.

[00032] The micro-organism capture apparatus 10 can be provided in a kit for taking a sample. As shown in TABLE 1 below, several components can be included in the kit.

TABLE 1

Additional equipment can be provided by the user of the apparatus, such as a peristaltic pump 34 that fits the tubing 32 (which is preferably 3.2 mm ID, 1.6 mm wall thickness), and a 25 L or less collection container 30, optionally with single use liners for easy reuse of the collection container 30. Optionally, this additional equipment can be provided in the kit. Sizes of each of the components in the kit can vary for different uses. Elements of the kit are manufactured according to standard methods. Use of the kit is further described in the example below.

[00033] There are several advantages to the present invention over the prior art. For example, the cost of the analysis of viruses is significantly less than the prior art. Samples can be concentrated in the field, eliminating the need for shipping high volume (100 - 1000 L) water samples to the laboratory for analysis. The assay can be performed in less than three hours, saving much time over the prior art. The small concentrated volumes obtained with the present invention allow for 100% of the concentrated sample to be easily analyzed, a significant improvement that improves the recovery and sensitivity over current methods.

[00034] Viruses and other micro-organisms are attracted and retained with the microorganism capture apparatus 10 as follows. Liquid is flowed through the micro-organism capture apparatus 10 in the inlet 14 and through the filter 18. The viruses or microorganisms within the liquid are attracted, retained, and concentrated with the attracting mechanism 20. The liquid then flows out of the outlet 16.

[00035] More specifically, the flowing step can be accomplished by pumping the liquid from a collection sample 30 (a smaller sample of a larger liquid source, or the liquid source itself) into the inlet 14, through the filter 18 and out of the outlet 16 of the microorganism capture apparatus 10. Preferably, the pump 34 is first set at a speed of 200 rpm, at least 10 L of the liquid is filtered, the pump speed is increased to 220 rpm for 30 seconds, the pump 34 is turned off, pressure is relieved by opening a pump head locking device 44, and tubing 32 is removed from a pump head 46. Next, tubing 32 can be removed from the micro-organism capture apparatus 10 by unscrewing a luer locking hose barb 38 on the inlet 14 and replacing with a luer lock end cap, the micro-organism capture apparatus 10 can be tapped at the outlet 16 and excess fluid removed, and a plug can be inserted at the outlet 16. A chain of custody form can then be filled out, the form and the micro-organism capture apparatus 10 can be inserted into return packaging, mailing supplies filled out, and the whole package can be returned to a laboratory for analysis. Essentially, this method can be performed with the kit described above. [00036] A fluidized bed can be created in the filter 18 as described above in order to retain the attracting mechanism 20 by performing dead-end filtration or pressurizing the filter 18 when the filter 18 is a hollow-fiber membrane. Multiple types of viruses or other micro-organisms as described above can be attracted and retained with this method. [00037] The present invention also provides a method of isolating viruses or other micro-organisms from a liquid sample, including the steps of flowing a liquid through the micro-organism capture apparatus 10 as described above, attracting, retaining, and concentrating the viruses or micro-organisms within the liquid with the attracting mechanism 20, releasing the attracting mechanism 20 from the filter 18 with the release mechanism 22, and isolating the viruses or micro-organisms retained by eluting the viruses or micro-organisms from the attracting mechanism 20. The flowing step can be performed as described above and with the kit as described above. A fluidized bed can be created in the filter 18 as described in the above method. Multiple types of viruses or other micro-organisms as described above can be attracted and retained with this method. Various assays can then be applied to the viruses isolated to determine what types of micro-organisms are present.

[00038] The present invention also provides a method of indicating that a liquid is need of remediation, including the steps of isolating at least one micro-organism from the liquid with the micro-organism capture apparatus 10, positively detecting the microorganism with an assay, and indicating that the liquid is in need of remediation. The isolating step can be performed according to the flowing step described above and with the kit as described above. Preferably, the liquid is water. Thus, the present invention is able to aid in water remediation and cleaning of sources of drinking water for future and continued use of those sources. This method allows for the determination of whether a large volume of water or liquid is contaminated by taking only a small sample of the water or liquid.

[00039] The invention is further described in detail by reference to the following experimental examples. These examples are provided for the purpose of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the present invention should in no way be construed as being limited to the following examples, but rather, be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

EXAMPLE 1

Sampling guidelines

[00040] 10 liters (~2.6 gallons) or greater of pool water sample should be taken for approximately every 25,000 gallons of water in the pool. Sample water from a depth of greater than 10" from the water's surface. In the case of a fecal accident, samples should be taken in close proximity to the accident. A back-flushed filter sample is also recommended.

[00041] While this example uses pool water, it is understood that any other liquid source can be used and can be collected in the same manner.

Instructions for use

1. Fill out Chain of custody form for the sample to be collected.

2. Remove micro-organism capture apparatus 10 from packaging (FIGURES 3 and 4A) and attach tubing at inlet 14.

3. Insert tubing into peristaltic pump system and close pump-head locking device (FIGURE 4B). Attaching the sample tubing to a pressurized line may be used in place of steps 3-8 (below). Do not collect samples at a pressure greater than 50psi.

4. Place tubing into sample.

5. Set peristaltic pump to a speed less than 275 rpm and turn the pump 'ON'

6. Filter the required 10 L (2.64 gal) volume of water (or more).

7. Before switching off pump, increase the pump speed 10% without stopping the pump for 30 seconds.

8. Switch 'OFF' the pump, open pump head locking device to relieve pressure, and remove tubing from pump head.

9. Remove the tubing by unscrewing the luer lock and replace with luer lock end cap. 10. Gently tap the micro-organism capture apparatus 10 at the filtrate exit point (outlet 16) to remove any excess fluid.

11. Insert a plug into the filtrate exit point (outlet 16).

12. Insert Chain of Custody Form and the micro-organism capture apparatus 10 into return packaging provided.

13. Fill out shipping Mailer and return to laboratory for assay analysis.

[00042] Throughout this application, various publications, including United States patents, are referenced by author and year and patents by number. The disclosures of these publications and patents in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.The invention has been described in an illustrative manner, and it is to be understood that the terminology which has been used is intended to be in the nature of words of description rather than of limitation.

[00043] Obviously, many modifications and variations of the present invention are possible in light of the above teachings. It is, therefore, to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described.

Claims

CLAIMSWhat is claimed is:

1. A micro-organism capture apparatus, comprising a housing including an inlet and outlet, said housing including therein a filter having attracting means for attracting and retaining micro-organisms, and release means for releasing said attracting means operatively attached to said housing.

2. The micro-organism capture apparatus of claim 1 , wherein said attracting means are chosen from the group consisting of positively and negatively charged particles.

3. The micro-organism capture apparatus of claim 2, wherein said attracting means are magnetic.

4. The micro-organism capture apparatus of claim 2, wherein said attracting means are nano-particles.

5. The micro-organism capture apparatus of claim 1 , wherein said attracting means include a positively charged coating.

6. The micro-organism capture apparatus of claim 5, wherein said positively charged coating is an antibody coating.

7. The micro-organism capture apparatus of claim 1 , wherein said attracting means further include means for attracting multiple types of viruses.

8. The micro-organism capture apparatus of claim 1 , wherein said filter is a hollow- fiber filter membrane.

9. The micro-organism capture apparatus of claim 8, wherein said release means is chosen from the group consisting of a removable cap, valve, seal, and plug-end.

10. The micro-organism capture apparatus of claim 1 , wherein said filter is a micro- porous membrane.

11. The micro-organism capture apparatus of claim 10, wherein said release means is a removable and disposable flat-bed filter.

12. The micro-organism capture apparatus of claim 1 , wherein said micro-organism is a virus is chosen from the group consisting of Caliciviruses (Noroviruses), Adenoviruses, and Polioviruses.

13. The micro-organism capture apparatus of claim 6, wherein said micro-organism is chosen from the group consisting of waterborne protozoa, Cryptosporidium parvum, and Giardia lamblia

14. The micro-organism capture apparatus of claim 1 , wherein said filter includes said attracting means pre-loaded therein.

15. The micro-organism capture apparatus of claim 1 , further including a pressure regulator valve operatively attached to a loop.

16. A kit for sampling a liquid for micro-organism, comprising the micro-organism capture apparatus of claim 1 , tubing, luer hose barb fittings, luer lock end caps, a plug, mailing supplies, and an instruction manual.

17. The kit of claim 16, further including a peristaltic pump and a collection container having a volume of less than 25 L.

18. A method of attracting and retaining micro-organisms, including the steps of: flowing liquid through the micro-organism capture apparatus of claim 1 ; and attracting, retaining, and concentrating micro-organisms within the liquid with the attracting means.

19. The method of claim 18, wherein said flowing step is further defined as flowing liquid into the inlet, through the filter, and out of the outlet of the micro-organism capture apparatus.

20. The method of claim 19, wherein said flowing liquid step is further defined as pumping the liquid from a collection sample into the inlet, through the filter and out of the outlet of the micro-organism capture apparatus.

21. The method of claim 20, wherein said pumping step is further defined as pumping at a speed of 200 - 300 rpm, filtering up to 25 L of the liquid, increasing the pump speed 10% for 30 seconds, turning the pump off, relieving pressure by opening a pump head locking device, and removing tubing from a pump head.

22. The method of claim 21 , further including the steps of removing tubing from the micro-organism capture apparatus by unscrewing a luer lock on the inlet and replacing with a luer lock end cap, tapping the micro-organism capture apparatus at the outlet and removing excess fluid, and inserting a plug at the outlet for safe transport.

23. The method of claim 22, further including the steps of filling out a chain of custody form, inserting the form and micro-organism capture apparatus into return packaging, filling out mailing supplies, and returning to a laboratory for analysis.

24. The method of claim 18, further including the step of creating a fluidized bed of the attracting means in the filter to be retained.

25. The method of claim 20, wherein said step of creating is further defined as containing attraction means by performing dead-end or restricted flow filtration.

26. The method of claim 20, wherein said step of creating is further defined as pressurizing the filter and wherein the filter is a hollow-fiber membrane.

27. The method of claim 18, wherein said attracting, retaining, and concentrating steps are performed for multiple types of micro-organisms.

28. The method of claim 18, wherein the micro-organisms are chosen from the group consisting of Noroviruses, Adenoviruses, polioviruses, Cryptosporidium parvum, Giardia lamblia, Bacillius anthraicis, and Vibrio cholera.

29. A method of isolating micro-organisms from a liquid sample, including the steps of: flowing liquid through the micro-organism capture apparatus of claim 1 ; attracting, retaining, and concentrating micro-organisms within the liquid with the attracting means; releasing the attracting means with the release means; and isolating the micro-organisms retained by eluting the micro-organisms from the attracting means.

30. The method of claim 29, wherein said flowing step is further defined as flowing liquid into the inlet, through the filter, and out of the outlet of the micro-organism capture apparatus.

31. The method of claim 30, wherein said flowing liquid step is further defined as pumping the liquid from a collection sample into the inlet, through the filter and out of the outlet of the micro-organism capture apparatus.

32. The method of claim 29, further including the step of creating a fluidized bed in the filter to retain the attracting means.

33. The method of claim 32, wherein said step of creating is further defined as performing dead-end filtration.

33. The method of claim 32, wherein said step of creating is further defined as pressurizing the filter and wherein the filter is a hollow-fiber membrane.

34. The method of claim 29, wherein said attracting, retaining, and concentrating steps are performed for multiple types of micro-organisms.

35. The method of claim 29, further including the step of analyzing the eluted microorganisms and determining the type of micro-organisms present.

36. The method of claim 29, wherein the micro-organisms are chosen from the group consisting of Noroviruses, Adenoviruses, polioviruses, Cryptosporidium parvum, Giardia lamblia, Bacillius anthraicis, and Vibrio cholera.

37. A method of indicating a liquid is in need of remediation, including the steps of: isolating at least one micro-organism with the micro-organism capture apparatus of claim 1 ; positively detecting the micro-organism with an assay; and indicating that the liquid is in need of remediation.

38. The method of claim 37, wherein the liquid is water.

39. The method of claim 37, wherein the micro-organisms are chosen from the group consisting of Noroviruses, Adenoviruses, polioviruses, Cryptosporidium parvum, Giardia lamblia, Bacillius anthraicis, and Vibrio cholera.