STABILIZED SALIVA TEST SYSTEM AND METHOD

A stabilized saliva testing system and method facilitates obtaining and preparing a stabilized saliva test sample for subsequent analysis. The system preferably includes a kit having various collection and preparation implements, 1× TE pH 8.0 as the stabilizer, a means for mailing the sample, and instructions for use. A user obtains a saliva sample, prepares the stabilized saliva test sample as instructed, and mails the sample to a 3rd party for analysis. The 3rd party tests the sample for tooth and gum health indicators using known testing instrumentation, and sends the user a report summarizing their tooth and gum health.

BACKGROUND OF THE INVENTION

The present invention relates to testing biological fluids, and more specifically to a system and method of testing stabilized saliva.

It is well established that oral health is extremely important. Oral infections such as tooth decay and gum disease can be unpleasant, unsightly and expensive. What is less known, however, is that there is a synergic relationship between oral health and general health. Gum disease, for example, is linked to a variety of illnesses including heart disease, diabetes, respiratory disease, osteoporosis, and rheumatoid arthritis. People with gum disease are twice as likely as others to die from a heart attack and three times as likely to have a stroke.

Testing biological fluids and tissues for a variety of conditions, diseases, markers and drugs is an integral part of health care. A blood panel, for example, is standard protocol in an annual physical examination. The analysis of saliva is less routine but can be extremely helpful in various situations. For example, saliva can be screened to help identify the presence of Cushing's disease, anovulation, HIV, cancer, parasites, hypogonadism and allergies, and for the presence of various compounds. Saliva can also be tested for tooth and gum health, and oral cavity cleanliness.

One of the barriers to testing biological fluids such as saliva is the availability of the requisite analysis tools. By way of example, it would be desirable for dentist offices to routinely offer saliva testing for monitoring oral health, but saliva testing instruments are beyond the reach for many practices. Likewise, the ordinary health-conscious consumer does not have access to the instrumentation necessary to monitor their oral health.

Another challenge associated with saliva analysis is specimen instability. Saliva is replete with microbes and proteolytic enzymes which alter the constituents of specimens over time, plus various biomarkers are affected by pH and temperature. For these and many other reasons, collecting and storing saliva specimens for later analysis is not simply a matter of spitting in a cup and testing it later.

As can be seen, there is a need for a saliva testing system and method that overcomes the shortcomings of the prior art. It is desirable that this system and method can facilitate sampling saliva in a non-clinical setting by non-trained personnel, stabilize that specimen for subsequent screening, analyze the specimen for various markers of oral and general health, and report those results. It is desirable that this system and method are reasonably easy to use, effectively stabilize saliva specimens, and yield reproducible and accurate screening data.

SUMMARY OF THE INVENTION

A saliva testing system facilitates obtaining a saliva sample and preparing a stabilized saliva test sample for subsequent analysis. The system preferably includes a kit having various collection and preparation implements, additives such as purified water and stabilizer, means for mailing the sample, and instructions for use. A user obtains a saliva sample, prepares the stabilized saliva test sample as instructed, and mails the sample to a 3rdparty for analysis. The 3rdparty tests the sample for tooth and gum health indicators using known testing instrumentation, and sends the user a report summarizing their tooth and gum health. The preferred stabilizer is 1× TE pH 8.0.

DETAILED DESCRIPTION OF THE INVENTION

Broadly, the present invention is a system and method of testing stabilized saliva whereby a user, typically a test subject, obtains a specimen of saliva, stabilizes it, mails it to a testing facility for analysis and receives a report of the results.

FIG. 1depicts an overview of the system and method with test subject20orally expressing saliva specimen24into collection cup32; inserting first and second specimen tubes37,38, and tracking card34into shipping pouch39; and mailing shipping pouch39to a testing facility for analysis by saliva testing instrument50.

FIG. 2depicts some specimen collection steps in saliva testing system10. Referring toFIG. 2A, swishing fluid33is introduced into empty collection cup32. In a preferred embodiment swishing fluid33is approximately 6 mL of sterile/purified water, preferably split between two 3 mL vessels. As shown inFIG. 2B, test subject20swishes swishing fluid33in their mouth for approximately 20 seconds. Saliva specimen24, which is a mixture of saliva22(not shown) and swishing fluid33, is then expelled into collection cup32, as shown inFIG. 2C. It is preferred that the saliva specimen is collected when the test subject wakes up in the morning, before brushing teeth or consuming food or drink in order to obtain a baseline of a subject's natural bacteria load and type. If this is not possible it is preferred that the test subject not brush or floss 30 minutes prior to collection and that they avoid food one hour prior to collection.

FIG. 3depicts some specimen preparation steps using saliva specimen24in collection cup32which has been collected as described inFIG. 2C. Saliva specimen24is preferably allocated for use in multiple analyses, withFIG. 3depicting a three-way division having a first allocation for deposition onto a tracking card, a second allocation for testing a specific set of biomarkers, and a third allocation for testing a different set of biomarkers. It should be understood that varying numbers of allocations are within the scope of this invention.

InFIG. 3Aan aliquot of saliva specimen24is removed from collection cup32. InFIG. 3Bthat aliquot is deposited onto reactive pad36of test strip35on tracking card34. Tracking card34preferably tests for pH and resistance to acid on the freshly collected sample. In a preferred embodiment tracking card34is photographed for subsequent submission to the testing facility, for example via a HIPAA-compliant text messaging system.

FIGS. 3C and 3Ddepict dispensing saliva specimen24from collection cup32up to a provided fill line (not shown) on first specimen tube37and adding stabilizer A40to yield first stabilized specimen42. AlthoughFIG. 3Cdepicts pouring saliva specimen24into first specimen tube37it is preferable to use a dropper such as transfer pipette31given it is generally more accurate to reach a fill line by dropping versus pouring. In a preferred embodiment first stabilized specimen42consists of approximately 50% stabilizer A and approximately 50% saliva specimen24by volume. Although not shown, first specimen tube37is preferably capped then inverted approximately 10 times to mix. In a preferred embodiment first stabilized specimen42is subsequently tested for cavity-causing bacteria, an indicator of tooth health.

FIGS. 3E and 3Fdepict dispensing saliva specimen24from collection cup32into second specimen tube38and adding stabilizer B41to yield second stabilized specimen43. Although not shown, for the reasons discussed above with respect toFIG. 3Cit is preferable to employ transfer pipette31to transfer saliva specimen up to fill line on specimen tube. In a preferred embodiment second stabilized specimen43consists of approximately 10% stabilizer and 90% saliva specimen24by volume. Second specimen tube38is preferably capped then inverted approximately 10 times to mix. In a preferred embodiment second stabilized specimen43is subsequently tested for inflammatory factor 1—red blood cells; inflammatory factor 2—white blood cells; and protein biomarkers, all of which are indicative of gum health.

It should be understood that stabilizer A40and stabilizer B41can be chemically identical although their proportional volume will likely vary in first stabilized specimen42versus second stabilized specimen43. By way of example, in a preferred embodiment first stabilized specimen42consists of 1 mL of stabilizer plus 1 mL of saliva specimen, thereby yielding a mixture of 50% stabilizer, while second stabilized specimen43consists of 300 μL of stabilizer plus 3 mL of saliva specimen, thereby yielding a mixture of 10% stabilizer.

In a preferred embodiment stabilizers A and B40,41are a TE buffer, with 1× TE pH 8.0 being most preferred. Among other sources this solution is commercially available as MB-040 from Rockland Immunochemicals.

Referring toFIG. 4, tracking card34, first stabilized specimen42and second stabilized specimen43, all prepared in accordance withFIG. 3, are put into shipping pouch39, which may be configured and labeled as a “BIOHAZARD” container. This is depicted inFIG. 4A. Shipping pouch39is mailed (FIG. 4B) to a testing facility having saliva testing instrument50. In a preferred embodiment first and second stabilized specimens42,43are loaded onto assay strip54and tested in accordance with standard testing protocols, as shown inFIG. 4C. Test report56is generated from said testing (FIG. 4D) and sent to test subject20and/or their designated agent. In a preferred embodiment saliva testing instrument50is modified and validated as a laboratory-developed test, with a commercially available but unmodified example being the SillHa Salivary Testing Instrument from ARKRAY USA, Inc., in Minneapolis, Minn.

In using a preferred embodiment of the present invention test subject20is provided the tools and information to collect and prepare their specimen without special training or equipment. This can be accomplished by providing testing kit30(not shown) including transfer pipette31, collection cup32, 2 vessels of 3 mL swishing fluid33, tracking card34, first specimen tube37, second specimen tube38, shipping pouch39, stabilizer A40, stabilizer B41and instructions for use. It is desirable that swishing fluid33, stabilizer A40and stabilizer B41are pre-measured so test subject20simply empties the contents of the associated vessel, for example “Stabilizer A” or “Stabilizer B” into the corresponding specimen tube37,38, without needing to measure specific volumes. As described above stabilizers A and B40,41may be the same chemical composition, although their associated vessels would likely contain different volumes.

The present invention is based on observations and data that indicate markers in unstabilized saliva change more than markers in stabilized saliva over time.

Ten oral rinse samples obtained at the same time from the same source were analyzed for five different markers, with two sample, A and B, per marker. They were analyzed at 0, 2, 4 and 6 hours using the SillHa Salivary Testing Instrument model LH-4912 in accordance with the associated standard protocol. The instrument measures oral rinse samples using the dual-wavelength reflectance method wherein the change in color of the test strip after sample application is measured by the reflectance of light with wavelengths of 565 nm, 635 nm and 760 nm. Results pertain to levels of cariogenic bacteria, acidity, buffer capacity, leukocyte, protein and ammonia, and are reported numerically as the Raw Value (0-100) and categorized as “low”, “average” or “high” based on the following numerical ranges:

Stabilized samples employ a different scale for “low”, “average” or “high” based on the SillHa Raw Value. These ranges are set forth below:

Table 1 depicts unstabilized Raw Values and associated categories, along with notations where the sample changed at least one category, or example from low to high, over the testing period:

It is noted that 60% of the samples demonstrated at least 1 category change over the 6-hour time period.

Table 3 reports the effect of time on the five markers in unstabilized samples over the course of five days using the same protocol.

As shown, 40% of the samples demonstrated at least 1 category change over the 6-day time period. It is notable that the 60% and 40% changes over 6 hours and 5 days, respectively, are not inconsistent. Rather, some data will curve over time, for example bacteria may initially increase in number then decline as nutrition is consumed, which would exhibit a curved versus linear growth pattern.

Table 5 reports the effect of time on the five markers in stabilized samples over the course of five days using the same protocol.

14% of the samples demonstrated at least 1 category change over the 6-day time period. These data suggest that saliva samples with stabilizer of the present invention are significantly more stable than unstabilized samples.

It should be understood that the foregoing relates to exemplary embodiments of the invention and that modifications may be made without departing from the spirit and scope of the invention as set forth in the following claims. Examples of modifications include the inclusion of additional markers and/or a totally “at home” system that does not require sending out a sample. Terms such as “substantially” and the like shall mean within reasonable bounds when considering limitations such as machines, materials, manufacturing methods, and people. By way of example, a “substantially smooth” surface means there are no intentional bumps or irregularities. All ranges set forth herein include the endpoints as well as all increments there between, even if not specifically stated. By way of example 1 to 2 inches includes 1 inch, 1.000001 inches and so forth. Finally, unless otherwise stated or contrary to common sense, “approximate” and the like shall mean +/−10%.