FIELD : The present invention relates to a device for the rapid detection and testing of diseases, disorders and immunity 'in humans and animals from a s-ample containing antibodies or antigens, as well as testing for disease-causing agents in the environment .

BACKGROUND

Antibodies are "natural" proteins produced by the body' s immune system in response to infecting micro-organisms such as bacteria or viruses. More specifically, each antibody is produced by lymphocytes, or specialized white cells, as a result of exposure to specific substances (mainly proteins and carbohydrates) called antigens, usually found on the outside of an invading organism. Antibodies are proteins, which are transported around the body in the plasma portion of the bloodstream. Antibodies are known to be shaped like the letter "Y" with a reactive site at the tip of each branch, ^• so that antibodies can become attached to antigens on the basis of their molecular shape. This can cause the infecting micro-organisms

to aggregate and become neutralized until they are taken away and dealt with by other white cells.

Antigens are large molecules (usually proteins) that act as "name tags" on the surface of cells, viruses, fungi, bacteria, and some non-living substances such as toxins, chemicals, drugs, and foreign particles and are considered "non-self" in the human body (e.g., not manufactured by the body). The immune system recognizes antigens as a threat to health and produces antibodies that attempt to destroy substances containing antigens identified by immune cells as dangerous pathogens in the body.

Some antibodies adhere to the surface of B cells (lymphocytes) , while others circulate throughout the body. When a circulating antibody attacks and grabs an antigen and its associated pathogen, the resulting combo-organism is called an antigen/antibody complex or immune complex. This complex is then engulfed by a phagocyte such as a macrophage, or filtered out of the blood by the immune system.

Antibodies can also attack antigens directly. At any given time the body has 10 million antibody shapes carried on the

surfaces of B cells ready to attack antigens in defense of the body. The following are the five main classes of antibodies:

Immunoglobulin A (IgA) - these protect mucous membranes and are present in salilva, tears 'and other secretions.

Immunoglobulin D (IgD) - rare and not tested due to no useful knowledge of function to date..

Immunoglobulin E (IgE) - their presence in elevated levels in the blood can be an indication of allergic reactions including hay fever, eczema and asthma.

Immunoglobulin G (IgG) - their presence is a sign of an ongoing or established infection.

Immunoglobulin M (IgM) - first antibody mobilized following an attack by a pathogen such as a virus, bacteria, fungus or toxin.

Testing for many diseases and disorders can take days, weeks and, in some cases even longer. In many instances such a delay is unacceptable. A second important factor in disease testing is that no test is viable unless it can offer a high degree of accuracy.

SUMMARY OF THE INVENTION

According to the invention there is provided a. device and method for testing for diseases, disorders and immunity in

humans, animals and the environment. The device includes a bottom membrane support assembly having a lower opening and operative to receive a bottom membrane extending over said lower opening and a top membrane support assembly having an upper opening and operative to receive a top membrane extending across said upper opening. The top membrane support assembly is positionable over the bottom membrane support assembly with the upper opening aligned with the lower opening.

The top and bottom membranes are aligned and parallel.

An absorbent pad may be located below the bottom membrane to absorb the various liquids that are poured over the membranes. Needless to say, the membranes are water permeable.

The bottom membrane support assembly may include a frame, an absorbent pad within said frame, and a bottom membrane mask plate having an opening, the opening positioned to overlay a membrane placed on the absorbent pad.

The top membrane support assembly may include an upper section, a top membrane support plate juxtaposed to a bottom of the upper section, and a lower section, the upper section, the top membrane support plate and the lower section all having

aligned openings, the upper section able to be positioned to overlay a top membrane which extends across the opening in both the upper section and the top membrane support plate.

The lower section has an opening in one side to permit sliding over and away from the bottom membrane support assembly.

The lower section may be hinged at one end to a hinge plate.

In another aspect of the invention there is provided a method of testing for diseases and disorders, which includes providing spaced -apart water permeable, parallel top and bottom membranes having deposited thereon desired reagents, such as antibodies or antigens and a protein marker, diluting a drop of sample liquid with a first liquid diluent, pouring the first diluent and dissolved sample over the top membrane and after being ^' filtered by the top membrane, over the bottom membrane and pouring a second liquid over said top membrane, and after being filtered by the top membrane, over the bottom membrane, to render visible any spots resulting from reaction with the sample by the reagents on the top and bottom membranes.

One of the membranes may be used as a control.

The membranes may be treated in localized areas with antigens and/or antibodies corresponding to up to 4 diseases or disorders .

The sample liquid may be a single drop of human blood.

BRIEF DESCRIPTION OF THE DRAWINGS

Further features and advantages will be apparent from the following detailed description, given by way ^' of example, of a preferred embodiment taken in conjunction with the accompanying drawings, wherein:

Fig. 1 is a perspective view of the testing device;

Fig. 2 is a perspective view of the device with the bottom portion exploded;

Fig. 3 is a perspective view of the device with the top portion exploded;

Fig. 4 is perspective view of an alternative embodiment of the device in which the -lower tray slides out of the side of the device with the lower portion exploded;

Fig. 5 is perspective view of the device of Fig-. 4 in which the lower tray is not in exploded form;

Fig. 6 is a perspective view of yet another embodiment which has the upper portion hinged to a base portion;

Fig. 7 is a perspective view of the embodiment of Fig. 6 in which a first diluent is being poured onto the first membrane;

Fig. 8 is a perspective view of the embodiment of Fig. 6 in which a marker fluid is being poured onto the first membrane; and

Fig. 9 is a perspective view of the embodiment of Fig. 6 in which a liquid used to cleanse the surface of the membrane is being poured onto the first membrane.

DETAILED DESCRIPTION WITH REFERENCE TO THE DRAWINGS Referring to Figs. 1-3, the testing device 10 has a top membrane support assembly 30 having an upper section 12 and a lower section 14. ' The upper section has a bowl-shaped recess 16 enclosing a circular opening 18. Below the upper section is a top membrane support plate 22 having an opening 24 identical in

size to and, when assembled, aligned with opening 18. At each corner there are holes 27 which are dimensioned to receive legs 20 extending down from upper section 12. Lower section 14 has a lower section plate 25, which also has a large opening 26 of the same diameter as openings 24 and 18 and is aligned with the latter openings when assembled. Lower section plate 25 also has holes 28 at each corner equal to in diameter and aligned with corresponding holes 27.

In Figs. 1-9, reference number 20 identifies a support plate on which the device 10 rests. The support plate 20 allows the lower section 14 to slide over the bottom membrane section 37 as shown in Figs. 4 and 5.

As seen in Figs. 2 and 3 a bottom membrane support assembly 37 has a rectangular opening 33 in a rectangular frame 32 having a ledge 42 around an interior periphery thereof, said rectangular opening 33 which receives an absorbent cotton inside of a plastic cartridge 34 and, on top of the absorbent cotton 34, a lower synthetic top membrane 36. Legs 38 of a bottom membrane mask plate 15 having a central opening 17 with the legs at each corner that pass through holes 40 and a bottom membrane mask plate 15 that rests atop ledge 42 and overlies top membrane

36 and absorbent cotton 34. The assembled bottom membrane section 37 is seen in Fig. 3.

Referring to Fig. 4, the rectangular frame 32, the absorbent cotton 34, the top membrane 36, and the bottom membrane mask plate 15 are shown in exploded form adjacent to the lower section 14, which has an opening 44 into which slides the bottom membrane section 37. This can be seen more clearly in Fig. 5.

An alternate embodiment having a hinge is shown in Fig. 6. In this case a hinge sleeve 48 is- integrally attached to an upper edge of plate 14. A pair of spaced apart hinge sleeve portions 50 are integrally attached to an upper edge of a hinge plate 51. With sleeve 48 aligned with sleeve portions 50, hinge pin 46 slides through the sleeve portions 50 and sleeve 48 locking the three together in a hinged connection. The remaining parts are the same as for the two embodiments discussed above. The device 10 is made of plastic both for ease of manufacture and cost.

Referring to Figs. 6-9, in use the device 10 is assembled with the top synthetic membrane 29 and the bottom synthetic membrane 36 in place. Each membrane 29 and 36 has deposited on

it in a given location either the desired antibodies or antigens for the test. The deposit on the synthetic membranes 29 and 36 are antibodies as well as a control, for example a protein- treated spot capable of capturing IgG antibodies normally present in blood and blood components. First a single drop of blood is transferred to the vial 52 which contains an inert diluent. Next the diluted blood sample is poured into the test device 10 where it reacts with the antibodies both on the top membrane 29 and on the bottom membrane 36 in this case producing circular dots or coloured regions. The control, for example a protein sens-itive only to human blood of the person being tested, reacts with the protein to produce a circular dot. If both the top membrane 29 and the bottom membrane 36 both show a control spot' then the test is valid. In the event that any of the other areas of the membranes 29 and 36 react to form a visible spot then the person being tested tests positive for whichever condition, disease or disorder is being tested. If the top and bottom membranes 29 and 36 are treated with antibodies or antigens corresponding to the same disease or disorder then the fact that spots show up on both membranes lends redundancy to the results and a consequent high reliability. Ordinarily the bottom membrane 36 is used as the control unit.

Alternatively, the top membrane 29 and bottom 36 membrane can be treated with antibodies or antigens corresponding to different diseases and/or disorders. In this case there can be up to four different diseases and/or disorders tested at one time (plus two controls), i For example,, the synthetic membranes 29 and 36 may be treated with antigens or antibodies corresponding to hepatitis, A, hepatitis B and Avian Flu (e.g., H5 antigen) . Potentially the apparatus can be used to test six different diseases, conditions or disorders at once, if no control is used.

Referring to Fig. 8, the second step is to pour the second liquid in vial 54 over the top and bottom membranes 29 and 36, respectively. This liquid causes the spots to become visible. Referring to Fig. 9, the final vial 56 is poured over the top and bottom membranes 29 and 36, respectively, to cleanse the surfaces of the membranes for a better showing of the spots. Both liquids are known in the industry.

The number of individual spots deposited in the membrane (i.e. the number of individual disease, condition or control spots) is limited only by the available space and sensitivity/concentration of the liquids being used. Although specific examples, wherein 4 or 6 conditions are tested at once

are discussed above, the number of "spots" used is ultimately up to the user.

The method may include applying a control spot, protein, antigen, etc. on one or both membranes. Alternatively, one or more of the spots, proteins, antigens, etc. may be duplicated on both membranes so as to provide redundancy.

Accordingly, while this invention has been described with reference to illustrative embodiments, this description is not intended to be construed in a limiting sense. Various modifications of the illustrative embodiments, as well as other embodiments of the invention, will be apparent to persons skilled in the art upon reference to this description. It is therefore contemplated that the appended claims will cover any such modifications or embodiments as fall within the true scope of the invention.