WO1994024557A1

WO1994024557A1 - Article and method for detecting the presence of pathogens in excreta

Info

Publication number: WO1994024557A1
Application number: PCT/US1994/004073
Authority: WO
Inventors: James G. Dobbins; Marion Koopmans; John A. Stewart; Philip E. Pellett
Original assignee: The United States Of America, As Represented By Th
Priority date: 1993-04-16
Filing date: 1994-04-13
Publication date: 1994-10-27
Also published as: AU6704294A

Abstract

An article and method for indicating potential affliction with pathologies that can be detected in excreta. The article comprises a means for detecting the presence of substances associated with such pathologies and a means for carrying the detecting means on a subject such that the detecting means can come in contact with the excreta. Methods for detecting infection with cytomegalovirus in excreta are also provided.

Description

ARTICLE AND METHOD FOR DETECTING THE PRESENCE OF PATHOGENS IN EXCRETA

BACKGROUND OF THE INVENTION

A need exists for a device or article that could be used to screen infants, newborns, and even adults, for a range of pathologies in a convenient and inexpensive manner. The present invention addresses that need by providing a means for rapid detection of multiple- pathologies that can be detecting in excreta.

Congenital infection with cytomegalovirus (CMV) is one such pathology that is the leading viral cause of birth defects, and a major cause of severe, multiple birth defects, in the United States. Approximately 40,000 infants, or 1% of all births, are congenitally infected each year; 4,000 of these infants are born with the severe defects that define the disease, and another 6,000 will have developmental problems later in life. The remaining 30,000 infected infants have no recognizable damage in the first few months of life, but may develop problems later that go unrecognized. Defects frequently noted at birth include microcephaly, intracranial calcifications, hearing and vision damage, and growth retardation. Developmental problems that may be noted in later include partial loss of vision or hearing, and learning disabilities.

Identification of the 90% of CMV-infected infants who are asymptomatic at birth is important to provide promising new treatments to block further viral damage, to implement testing to identify subtle or late damage to these infants, and to fully characterize the epidemiology of infection and transmission of the virus during pregnancy. The only method of detecting CMV infection currently in use is culture of urine. This procedure is unsuitable because it requires special handling, is typically expensive and the results are not known for two or three days after a child leaves the newborn nursery.

From a public health perspective, this existing test is not suitable for national screening of all newborns. A detection method is needed that is inexpensive, easy to use in all types of medical settings, will produce rapid results, and does not require expensive equipment or staff training.

Assays for the detection of viral proteins, antibodies against viruses, or other proteins that may be associated with viral infection, are provided herein. Incorporation of those alternative methods is provided herein, thereby addressing the existing need.

SUMMARY OF THE INVENTION

This invention provides an article that would be worn by a person for the purpose of indicating potential affliction with pathologies that can be detected in excreta. The article comprises a means for detecting the presence of substances associated with such pathologies and a means for carrying the detecting means such that the detecting means can come in contact with the excreta. Embodiments of the carrying means include a diaper, a tampon and a feminine hygiene pad. The detecting means can either be integrally found on or in the carrying means (as, for example, disposed on the fibrous material of a diaper) or disposed on a substrate that is detachably secured to the carrying means. The substrate can be constructed of a non-porous, flexible material, such as plastic, which is in the form of a strip. The strip is capable of being placed within a pocket formed on the carrying means so that after it has come in contact with the excreta, the strip can be removed for analysis of the detecting means.

The detecting means can be an antigen, such as a human virus or an antigen of the virus, or an antibody. The virus can be cytomegalovirus (CMV) or human immunodeficiency virus (HIV) . The substance detected can be a protein, glucose, bilirubin, a ketone, hemoglobin, urobilinogen, a nitrite, a leukocyte or a combination thereof.

Methods of detecting CMV or substances associated with current or previous CMV infection in excreta are also provided. A method of diagnosing a subject as potentially afflicted with such pathologies and a method of analyzing the detecting means after it has been in contact with excreta are also provided.

BRIEF DESCRIPTION OF THE FIGURES OF THE DRAWING

FIG. 1 is a perspective view of the absorptive side of a disposable diagnostic diaper illustrating a possible location for positioning the detecting means of the present invention on the diaper;

FIG. 2 is a cross-sectional view of the disposable diagnostic diaper taken along Line 2—2 in FIG. 1;

FIG. 3 is a cross-sectional view of an alternative embodiment of a diaper illustrating that the detecting means can be interspersed among the fibers of the absorptive portion the diaper;

FIG. 4 is a perspective view of a substrate of the type that is contained within the retaining means of the diaper in FIG. 1;

FIG. 5 is a perspective view of a tampon with the detecting means thereon or embedded therein;

FIG. 6 is a perspective view of an alternative embodiment of a tampon with the detecting means interspersed among the fibers of the absorptive portion the tampon;

FIG. 7 is a perspective view of a feminine hygiene pad with the detecting means of the present invention therein; and

FIG. 8 is a perspective view of an alternative embodiment of a feminine hygiene pad with the detecting means interspersed among the fibers of the absorptive portion the pad. DETAILED DESCRIPTION OF THE ILLUSTRATIVE EMBODIMENTS

Referring to FIG. 1, a diaper 10 is shown as a means for carrying the means for detecting at least one substance associated with at least one pathology that can be detected in excreta. The diaper 10 may be of the disposable type. A retaining means in the form of a pocket or sleeve 12 on the absorptive side 14 of the diaper receives at least one substrate 16 containing thereon, or embedded therein, a detecting means.

"Detecting means" as used herein would include as many particles, be they enzymes, antibodies or antigens, as would be necessary to detect the substance associated with the pathology being diagnosed. The substrate 16 is normally of a flexible construction which would be in fluid communication with the excreta of the user through the pocket 12 which may be constructed of a porous material. The pocket 12 may be integrated into the diaper 10, or may be detachably secured thereon.

The detecting means 18 on a single substrate 16 could be specifically reactive to only one substance associated with a pathology that can be detected in excreta. Alternatively, the substrate 16 could have thereon a plurality of detecting means 18 for detecting the presence of a plurality of pathogens.

The previously available combination of detecting means and substrate for determining the presence of, among other pathology related substances, ketone bodies, glucose, protein, and blood (Fraser, J. et al.: Studies With a Simplified Nitroprusside Test for Ketone Bodies in Urine, Serum, Plasma and Milk, Clin. Chim. Acta., 11:372-378 (1965); Ames^® Reagent Strips package inserts, Miles, Inc., Diagnostics Division (February 1992)), have the disadvantage of the additional step of urine collection. The present invention obviates the need for separate collection since the detecting means comes directly in contact with excreta in situ.

A substance "associated with" a pathology can be a metabolite or other chemical not normally found in the type of excreta being tested, or not normally found in the type of excreta being tested in a given concentration that is clinically relevant and not normally found in the excreta of healthy subjects. The individual detecting means 18 could be specifically reactive with a selected one of many "substances", including antigens, antibodies, nucleic acids, amino acids, proteins, glucose, bilirubin, ketone bodies, hemoglobin, urobilinogen, nitrites and leukocytes, or any other substance that is associated with a pathology that can be detected in excreta. As used herein, the term "antigen" includes antigenically specific fragments thereof. "Excreta" is any excretion of waste from an animal body, namely, urine, feces or menstrual discharge.

The term "reactive" as applied to the present detecting means 18 capable of binding or otherwise associating nonrandomly with a substance.

"Specifically reactive" as used herein to describe an antigen or antibody detecting means describes an antigen or antibody that does not react substantially with any pathology associated substance (antibody or antigen, respectively) other than the one specified, e.g. CMV antigen or antibody. When the detecting means is an enzyme, enzyme substrate or metabolite, "specifically reactive" describes an enzyme, enzyme substrate, metabolite etc. that does not substantially stearically associate with or bind to any pathology associated substance (metabolite, enzyme etc.) other than the one specified, e.g. bilirubin, glucose etc. "Detecting means" includes the antigen, antibody, enzyme, or metabolite etc. that is specifically reactive with the pathology associated substance as well as other reagents used for capturing the pathology associated substance and for the analysis of the detecting means to determine the presence of the pathology associated substance. The capturing means can be the specifically reactive detecting means or it can be a reagent capable of binding a class of compounds, such as proteins, which can then be analyzed more specifically.

When amino acids, proteins, glucose, bilirubin, ketone bodies, hemoglobin, urobilinogen, nitrites and leukocytes are being detected, an enzyme would likely mediate the specific reaction between the detecting means 18 and the substance detected. In such cases, the enzyme illustrations of such enzyme-mediated reactions are found in Examples I-VII below.

When viruses, such as HIV or CMV are being detected, antigen-antibody forces would likely mediate the specific reaction between the detecting means and the virus. The specific reaction could be between an antibody (detecting means 18) bound to the substrate 16 and pathology associated antigens or other antibodies in the excreta. The specific reaction can also be between an antigen (detecting means 18) bound to the substrate 16 and pathology associated antibodies in the excreta. It is also possible to use a detecting means, antibody or antigen based, that is specifically reactive with more than one pathology associated substance.

When the pathology associated substance is measured by an antigen enzyme immunoassays such as immunofluorescence assays (IFA) , enzyme linked immunosorbent assays (ELISA) and immunoblotting the diaper insert can be readily adapted to accomplish the detection of the antigen. An ELISA method effective for the detection of the antigen can, for example, be as follows: (1) bind the antibody to the diaper insert; (2) contact the bound antibody with excreta fluid containing the antigen; (3) contact the above with a secondary antibody bound to a detectable moiety (e.g., horseradish peroxidase enzyme or alkaline phosphatase enzyme) ; (4) contact the above with the substrate for the enzyme; (5) contact the above with a color reagent; (6) observe color change. The above method can be readily modified to detect pathology associated antibody as well as antigen.

Examples of methods for detecting a an antigen associated with cytomegalovirus is presented in Example VIII. One method details the detection of anti-CMV antibody using a modified Western Blot protocol. A second method details the detection of CMV itself or a substance associated with CMV infection, after dissociation from the complex formed by the subject's own anti-CMV antibody, using a modified ELISA protocol.

Another immunologic technique that can be useful as a detecting step utilizes monoclonal antibodies (MAbs) for detection of antibodies specifically reactive with the antigen to be detected. Briefly, excreta from the subject is reacted with the antigen bound to a substrate (e.g. an ELISA 96-well plate) . Excess fluid is thoroughly washed away. A labeled (enzyme-linked, fluorescent, radioactive, etc.) monoclonal antibody is then reacted with the previously reacted antigen-serum antibody complex. The amount of inhibition of monoclonal antibody binding is measured relative to a control (no patient serum antibody) . The degree of monoclonal antibody inhibition is a very specific test for a particular variety or strain since it is based on monoclonal antibody binding specificity. MAbs can also be used for detection directly in cells by IFA. A micro-agglutination test can also be used to detect the presence of the pathology associated substance in the excreta of the subject. Briefly, latex beads (or red blood cells) are coated with the antigen and mixed with a sample of excreta from the subject, such that antibodies in the excreta that are specifically reactive with the antigen crosslink with the antigen, causing agglutination. The agglutinated antigen-antibody complexes form a precipitate, visible with the naked eye or by spectrophotometer. In a modification of the above test, antibodies specifically reactive with the antigen can be bound to the beads and antigen in the excreta thereby detected.

In addition, as in a typical sandwich assay, the antibody can be bound to a solid phase and reacted with the antigen. Thereafter, a secondary labeled antibody is bound to epitopes not recognized by the first antibody and the secondary antibody is detected. Since the present invention provides examples of current or previous pathology associated antigens other methods such as flow cytometry and immunoprecipitation can also be used as detection methods.

In the diagnostic methods taught herein, the antigen or antibody can be bound to a solid phase and contacted by a fluid sample of excreta. This sample can be taken directly from the patient or in a partially purified form. In this manner, antibodies specific for the antigen (the primary antibody) will specifically react with the bound antigen. Thereafter, a secondary antibody bound to, or labeled with, a detectable moiety can be added to enhance the detection of the primary antibody. Generally, the secondary antibody or other ligand which is reactive, either specifically with a different epitope of the antigen or nonspecifically with the ligand or reacted antibody, will be selected for its ability to react with multiple sites on the primary antibody. Thus, for example, several molecules of the secondary antibody can react with each primary antibody, making the primary antibody more detectable.

The detectable moiety will allow visual detection of a precipitate or a color change, visual detection by microscopy, or automated detection by spectrophotometry, radiometric measurement or the like. Examples of detectable moieties include fluorescein and rhoda ine (for fluorescence microscopy) , horseradish peroxidase (for either light or electron microscopy and biochemical detection) , biotin-streptavidin (for light, electron microscopy or biochemistry) and alkaline phosphatase (for biochemical detection by color change) . The detection methods and moieties used can be selected, for example, from the list above or other suitable examples by the standard criteria applied to such selections (Harlow and Lane, 1988) .

When nucleic acids are being detected the detecting means 18 on the substrate 16 can be a nucleic acid complimentary to a nucleic acid associated with the pathology. The complimentary nucleic acid in excreta will selectively hybridize with the detecting means which can then be analyzed for the presence of the pathology associated nucleic acid. Standard nucleic acid detection methods including, but not limited to, polymerase chain reaction (PCR) , restriction fragment length polymorphisms (RFLP) etc. can be applied (Sombrook et al.) to determine the source of the nucleic acid.

By positioning a plurality of substrates 16 within the retaining means 12 such that the substrates 16 and detecting means 18 thereon, or embedded therein, come in contact with excreta, the presence of many substances associated with a plurality of pathologies can be detected through a single exposure to excreta.

Alternatively, the detecting means 18 could be embedded in the diaper 10 itself, thereby obviating the need for a separate substrate 16. FIG. 3 discloses such an embodiment of a diaper 10 that indicates that the detecting means 18 are interspersed within or on the fibrous material 26 from which the diaper 10 is constructed, rather than being placed on a separate substrate 16.

Shown in FIG. 2 is a cross sectional view of the pocket 12 with a substrate 16 in place therein. The pocket 12 is shown as enclosing the substrate 16 between two members. A porous first member 20 is exposed on one side to the skin of the subject and on the other side to the/lumen 22 of the pocket 12, such that the substrate 16 comes in contact with excreta through the first member 20 which is acting as a passageway for the excreta. A second member 24 is exposed on one side to the lumen 22 of the retaining means 12 and the other side to either some layer of the diaper composition or the outer surface of the diaper 10. The second member 24 may be comprised of a substance such as a plastic that would be sufficiently translucent, and perhaps even transparent, to act as a window to allow one to view the substrates 16 through the second member 24 from outside the diaper 10.

FIG. 4 depicts the substrate 16, in the form of a strip, with the plurality of detecting means 18 thereon or embedded therein.

Another embodiment of the present invention is shown in FIG 5 in which the carrying means is a tampon 28 with a plurality of detecting means 18 contained on or embedded in a substrate 30. The substrate 30 can be permanently affixed to the tampon 28 or can be detachably secured thereon, as perhaps with a VELCRO^® attachment.

FIG. 6 shows another embodiment of the present invention in which the carrying means is a tampon 28 with a plurality of detecting means 18 contained on or embedded in the material of the tampon 28 itself.

Shown in FIG. 7 is a further embodiment of the present invention in which the carrying means is a feminine hygiene pad 32. A plurality of substrates 16, each having thereon a plurality of detecting means 18 are contained in a retaining means 34, such as a pocket. The substrates 16 can be removed for further analysis.

FIG. 8 discloses an alternative embodiment in which the carrying means is a feminine hygiene pad 32. A plurality of detecting means 18 are shown as being directly on or embedded in the fibers of the pad 32 itself.

The following examples are intended to illustrate but not limit the invention.

EXAMPLE I

DETECTION OF KETOSIS

The present invention is used to assess the severity of human ketosis by detecting the presence of ketone bodies in urine. The substrate used is a strip impregnated with reagents capable of detecting ketone bodies. Sodium nitroprusside, glycine and alkaline phosphate buffers are the primary ingredients. A pH of 9 is provided which is optimal for the reaction of nitroprusside with acetoacetic acid and acetone. The concentration of nitroprusside is approximately 7 % of the dry weight at time of impregnation. Visually assessable positive reactions are uniform and range from bright lavender to bright purple. Ketone results are interpreted as negative, small, moderate or large depending on the resulting color.

EXAMPLE II DETECTION OF BILIRUBIN

The present invention is used to assess the existence of liver disease, such as viral hepatitis, by detection of even very low concentrations of bilirubin in urine. Normally no bilirubin is detectable in urine by even the most sensitive of methods. Trace amounts in the urine are thus sufficient to warrant further testing.

The substrate used is a strip to which detecting means (reagents) specifically reactive with bilirubin are affixed. The detection is based on the coupling of bilirubin with diazotized dichloroaniline in a strongly acidic medium. Reagents include 2,4-dichloroaniline, diazonium salt and buffers. In the analysis step the substrate having detecting means thereon is contacted with diazonium salt resulting in the appearance of color. The color ranges through various shades of tan.

EXAMPLE III DETECTION OF HEMOGLOBIN

The present invention is used to detect even low concentrations of blood in the urine. The significance of trace amounts of blood in the urine may vary between patients but the existence of blood in urine warrants further testing.

The substrate used is a strip to which specific reagents reactive with hemoglobin or myoglobin are affixed. Reagents include 6.8% diisopropyl benzene dihydroperoxide, 4.0% 3,3',5,5' tetromethylbenzidine and 48% buffers. The detection is based on the peroxidase- like activity of hemoglobin, which catalyzes the reaction of diisopropylbenzene dihydroperoxide and 3,3' ,5,5'- tetramethylbenzidine. The resulting color ranges from orange through green. Further assessment and microscopic inspection may be warranted by the presence of green spots or a uniform green color. Reactions ranging from trace to large, with an equally broad range of occurrence of spots, may be observed.

Hemoglobin concentration of 0.015-0.062 mg/dl is equivalent to 5-20 intact cells per microliter of urine. The sensitivity of this test may be reduced in urine with high specific gravity.

EXAMPLE IV DETECTION OF PROTEIN

No protein is detectable in normal urine. Therefore, any amount of protein detected beyond a trace amount would indicate proteinuria. Proteinuria can be caused by a host of disorders including cardiac disease, intestinal decomposition, persistent fever, hyperthyroidism, or multiple myeloma, but is usually indicative of renal disease or damage.

The substrate used is a strip to which detecting means specifically reactive with protein are affixed. Reagents include 0.3% tetrabromophenol and 97.3% buffers. The reagents used would be more sensitive to albumins than globulins. The reaction is based on the protein-error-of- indicators principle. That is, at a constant pH, any green color indicates the presence of protein. Colors range from yellow for negative to blue for positive.

EXAMPLE V MEASUREMENT OF UROBILINOGEN

Urobilinogen is a colorless compound formed in the intestines by the reduction of bilirubin. It may be oxidized to urobilin in urine, where high concentrations indicate cirrhosis of the liver. The substrate used is a strip to which detecting means specifically reactive with urobilinogen would be affixed. The test is based on a modified Ehrlich reaction in which 0.2 % p- diethyla inobenzaldehyde in conjunction with a color enhancer reacts with urobilinogen in a highly acidic medium to form a pink-red color.

The normal range of urobilinogen concentrations detected with this method is 0.2-1.0 mg/dl (1 mg/dl is approximately equal to one Ehrlich unit/dl) . A result of 2.0 mg/dl represents the transition from normal to abnormal.

EXAMPLE VI DETECTION OF NITRITE

This reaction depends on the conversion of nitrate to nitrite by Gram-negative bacteria in the urine. The test is, therefore, a reliable predictor of bacteriuria and the attendant urinary tract infection. The substrate used would be a strip to which detecting means specifically reactive with nitrite would be affixed. At an acid pH, nitrite reacts with p-arsanilic acid to form a diazonium compound. This compound couples with 1,2,3,4- tetrahydrobenzo(h)-quinolin-3-ol to produce a pink color. Reagents include 1.4% p-arsanilic acid, 1.3% 1,2,3,4- tetrahydrobenzo(h)-quinolin-3-ol and 10.8% buffers. Any degree of uniform pink coloration would indicate the presence of 10⁵ or more organisms per ml.

EXAMPLE VII DETECTION OF LEUKOCYTES

The detection of leukocytes in the urine indicates the condition known as pyuria. Normal urine produces negative results in the following test and any positive result, small to large, is clinically significant.

The substrate used is a strip to which detecting means specifically reactive with leukocytes are affixed. Reagents include 0.4% derivatized pyrrole amino acid ester, 0.2% diazonium salt and 40.9% buffers. The test is based on the principle that leukocytes contain esterases that catalyze the hydrolysis of derivatized pyrrole amino acid ester to liberate 3-hydroxy-5-phenyl pyrrole. This product then reacts with a diazonium salt to produce a purple product.

EXAMPLE VIII DETECTION OF CYTOMEGALOVIRUS ANTIBODIES

ELISA for the detection of CMV antigen.

Dissociation of the CMV-antibody complex that may exist in urine is accomplished by adding .12 M hydrochloric acid (HCl) to the urine sample and incubating for 30 min. The urine is then neutralized to pH 7 using .5M HEPES buffer.

ELISA Plates are coated overnight at 4°C with lOOμl of 1:400 dilution of CMV-Ig that had been prepared from serum of an immunized goat by ammoniumsulphate precipitation and affinity purification. All incubations are done for 1 h at 37°C, and the plates are washed between each step with phosphate buffered saline solution with 0.05% Tween 20, pH 7.4 (PBSST) . After the overnight coating step, the plates are incubated with 5% milk in PBSS, dissociated antigen in 40 mM HEPES buffer with 5% milk (HM) , a 1:3000 dilution of two monoclonal antibodies to a late CMV antigen (MAB8125 and 8126, Chemicon Int., Temecula, CA) in HM, alkaline phosphatase labelled rabbit- anti-mouse Ig (Tago, Burlinghame, CA) in HM, and the enzyme substrate. This method can detect CMV present in urine.

Western blot (WB) assays for CMV antibodies. Crude lysates are prepared from overnight cultures of Escherichia Coli expressing the major antigenic protein ppl50 of CMV (kindly provided by Dr. D. Sanchez, CDC) . Alternatively, CMV grown in culture, which is commercially available, ATCC No. ATCCL136 VR538, can be used as an antigen. The proteins are separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose by electroblotting. After cutting the membrane into strips, it is blocked by incubation for 1 h at RT with 5% milk and 3% bovine serum albumin in PBSS (PBSSMB) , followed by a wash (3 x 5') with PBSST. All incubation are followed by a washing step and are done in 400μl volumes at room temperature (RT) for 1 h, except the substrate incubation which is 3-4 h long. Consecutively, 400μl of a 1:4 dilution of urine or a 1:200 dilution of control sera in HM are added, and bound antibody is detected by a 1:500 dilution of alkaline phosphatase labelled goat antibody to human IgG (Kirkegaard and Perry, Gaithersburg, MD) in PBSSMB, followed by a rinse in PBSS and incubation with the enzyme substrate. Virus specificity of the WB results is tested by blocking assays, in which the urine or serum samples are incubated before the actual WB assay with a predetermined concentration of cell culture-grown CMV (ATCC No. ATCCL136 VR538) , HHV6, or the respective uninfected cell lysates as controls. To rule out non¬ specific binding of Ig to Fc-receptors, or the unlikely possibility of B2m mediated conjugate binding, the samples are preincubated with serum from a CMV-seronegative individual or with a high concentration of purified B2m.

Claims

What is claimed is:

1. An article for indicating potential affliction with at least one pathology associated with a substance that can be detected in the excreta of a subject, comprising: means for detecting the presence of at least one substance in the excreta of the subject associated with at least one of the pathologies; and means for carrying the detecting means on the subject such that the detecting means can come in contact with the excreta.

2. The article of Claim 1, wherein the carrying means has at least a portion thereof comprised of an absorptive material.

3. The article of Claim 2, wherein the detecting means is on the absorptive material.

4. The article of Claim 1, wherein the detecting means is detachably positioned on the carrying means.

5. The article of Claim 1, wherein the carrying means is a diaper.

6. The article of Claim 5, further comprising a substrate to which the detecting means is bound and wherein the diaper further comprises a means for retaining the substrate thereon.

7. The article of Claim 1, wherein the carrying means is a tampon.

8. The article of Claim 1, wherein the carrying means is a feminine hygiene pad.

9. The article of Claim 6, wherein the carrying means has at least a portion thereof comprised of an absorptive material and wherein the retaining means comprises a passageway formed in the absorptive material that receives therein the substrate with the detecting means thereon.

10. The article of Claim 1, wherein the detecting means comprises an antigen.

11. The article of Claim 10, wherein the antigen is a virus or an antigen of the virus.

12. The article of Claim 11, wherein the virus is cytomegalovirus.

13. The article of Claim 11, wherein the virus is human immunodeficiency virus.

14. The article of Claim 1, wherein the detecting means comprises an antibody.

15. The article of Claim 14, wherein the antibody is specifically reactive with a virus.

16. The article of Claim 15, wherein the virus is cytomegalovirus.

17. The article of Claim 15, wherein the virus is human immunodeficiency virus.

18. The article of Claim 9, wherein the substrate is formed from a plastic material.

19. The article of Claim 1, wherein the substance detected is selected from the group consisting of: protein, glucose, bilirubin, ketone, hemoglobin, urobilinogen, nitrite and leukocyte.

20. A method of diagnosing a pathology associated with a substance that can be detected in excreta, comprising the steps of: a. disposing a carrying means, with a detecting means thereon that is specifically reactive with at least one substance that is associated with the pathology, on the body of a subject such that the detecting means comes in contact with the excreta; and b. analyzing the detecting means after it has come into contact with the excreta for determining the presence of the substance or amount thereof, the presence or amount of the substance indicating the presence of the pathology.

21. A method of diagnosing a pathology associated with a substance that can be detected in excreta, comprised of the step of: incorporating a detecting means on a carrying means that is disposed on the body of a subject, whereby the detecting means can be analyzed after it has come in contact with the excreta for determining the presence of at least one substance that is associated with a pathology or the amount of the substance present, the presence or amount of the substance indicating the presence of the pathology.

22. The method of Claim 21, wherein the carrying means is comprised of fibers and the incorporating step comprises the step of placing the detecting means on at least a portion of the fibers.

23. The method of Claim 21, wherein the incorporating step comprises the steps of placing the detecting means on a substrate and detachably incorporating the substrate on the carrying means.

24. A method of diagnosing a pathology that can be detected in excreta comprising the steps of: analyzing a detecting means that has been on a carrying means disposed on the body of a subject so that the detecting means has been in contact with excreta in order to determine the presence of at least one substance associated with the pathology or the amount of the substance, the presence or amount of the substance indicating the presence of the pathology.

25. An article for indicating potential affliction with cytomegalovirus, comprising: means for detecting the presence of cytomegalovirus or at least one substance associated with cytomegalovirus infection in the excreta of a subject; and means for carrying the detecting means on the subject such that the detecting means can come in contact with the excreta.

26. The article of Claim 25, wherein the excreta is urine.

27. A method of diagnosing current or previous cytomegalovirus infection in a subject, comprising the steps of: a. disposing a carrying means, with a detecting means thereon that is specifically reactive with cytomegalovirus or at least one substance that is associated with cytomegalovirus infection on the body of the subject such that the detecting means comes in contact with the excreta; and b. analyzing the detecting means after it has come into contact with the excreta for determining the presence of cytomegalovirus or a substance associated with cytomegalovirus infection or an amount thereof, the presence or amount of the substance indicating the presence of the pathology.

28. The method of claim 27, wherein the step of analyzing the detecting means comprises the steps of: contacting the detecting means with a detectable amount of an antibody specifically reactive with cytomegalovirus or an antigen thereof; and detecting the reaction of the antibody and the cytomegalovirus or antigen thereof on the detecting means, the reaction indicating current or previous cytomegalovirus infection in the subject.

29. The method of Claim 28 wherein the excreta is urine.

30. The method of claim 27, wherein the step of analyzing the detecting means comprises the steps of: contacting the detecting means with a detectable amount of cytomegalovirus or an antigen thereof; and detecting the reaction of the cytomegalovirus or antigen with an antibody reactive therewith on the detecting means, the reaction indicating current or previous cytomegalovirus infection in the subject.

31. The method of Claim 30, wherein the excreta is urine.

32. A method of diagnosing cytomegalovirus infection in a subject, comprising the steps of: contacting a urine sample from the subject with a detectable amount of an antibody specifically reactive with cytomegalovirus or an antigen thereof; and detecting the reaction of the antibody and the cytomegalovirus or antigen in the urine, the reaction indicating cytomegalovirus infection in the subject.

33. A method of diagnosing current or previous cytomegalovirus infection in a subject comprising the steps of: contacting a urine sample from the subject with a detectable amount of cytomegalovirus or an antigen thereof; and detecting the reaction of the cytomegalovirus or antigen with an antibody in the urine, the reaction indicating current or previous cytomegalovirus infection in the subject.