Patent Application
20230175042
Non-Applicable
The present invention relates generally to a means of detecting a target molecule, compound or substance through target-specific attraction with a dual-labeled probe whereby said probe expresses a high affinity for a distinct target and is an “excitable”, fluorophore capable of luminescence in a determinable light wave range. Specifically, the oligonucleotide sequence defining the target-specific binding site, takes the form of a dual-labeled probe forming a quenched, stem-loop structure in its native state which fluoresces upon hybridization to a target molecule nucleotide sequence. Said hybridization allows for optical detection of localized probe-conjugate combinations through a directed, filtered and focused light source. In the presence of a target molecule, nano-molecular probe-target conjugates illuminate and are made both identifiable (qualifiable) and quantifiable via luminescence (i.e., light wave excitation) and subsequent photo detection. The presence and quantity of the target molecule may thereby be both detectable and quantifiable through improved optics utilizing ellipsoid reflection enhancing both sensitivity and specificity.
The ability to detect and localize (i.e., qualify), monitor and quantify the expression of specific viral gene expressions in real-time offers unparalleled opportunities for advancements in molecular biology, disease pathophysiology, disease detection and medical diagnostics. Nonetheless, methods currently employed (e.g., selective amplification through multistep Polymerase Chain Reaction sample amplification and “saturation binding” in the case of a microassay such as ELISA) achieve variable rates of sensitivity and specificity with defined shortcomings in their applicability and practicality.
Specifically, in the case of PCR a particular sample must undergo a denaturization (i.e., separation) and synthesis process, with required sequential variations in temperature, for successful completion of each respective step which ultimately results in millions to billions of replicated copies. This process may be repeated on the order of 20 to 40 times (cycles) through the beginning stages of sample procurement, into exponential amplification and sequential thermal cycling resulting in sample isolation and magnification. And while this process is undeniably advantageous in terms of its sensitivity and ability to quantify the amount or number of a specific antigen molecules, the process is also limiting in that (1) the PCR method is multi-step and time-dependent (i.e., non-rapid) thus requiring highly trained personnel and sophisticated and dedicated equipment for sample processing where (2) the target sequence must be first determined and (3) mutations and contaminations are an underlying and ongoing concern.
Currently, in the case of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the identity of the etiological causative agent of coronavirus disease 2019 (COVID-19) is accomplished through a variation of PCR, real-time reverse transcription-polymerase chain reaction (RT-PCR). Through the testing of nasopharyngeal and oropharyngeal fluids, virus shedding is found to be in largest concentrations 5-7 days after symptom onset and continuing in peak concentrations for several weeks after onset. Functionally, swab-derived viral RNA is isolated and converted to complementary DNA (cDNA) which is then amplified (through cycling as above) using Taq DNA polymerase and a series of specific heating and cooling (thermo cycling) events. Manifestly, RT-PCR remains the definitive means of detection, owning to evidenced proven sensitivity, precision and specificity for viral detection over and above available serological tests. But this specificity comes at a cost of (a) time, wherein turnaround time can take from hours to days, and (b) immobility, test being commonly performed in a laboratory setting. Other limitations in RT-PCT include sample storage requirements, sample contamination and, ultimately, cost.
Serological microassays (e.g., ELISA, EIA (enzyme immunoassay) and RT-LAMP (loop-mediated isothermal amplification) in saliva) are a commonly utilized analytical assay for qualitatively and quantitatively measuring and assessing the presence of a target entity within a sample by implementing qualitative detection of IgG or IgM antibodies. Such tests may verify or accompany RT-PCR results, determine a previous immune response against the viral spike (S) protein, include a quantitative aspect and prove helpful to assess acquired immunity against subsequent viral exposure, a basis for assessing reinfection and/or for contact tracing purposes. And while these assays have proven indispensable in modern medical technology, the detection of target molecules is in no way simple. The process entails a multi-step process consisting of pre- and post-analytic steps exclusive of the actual analytic steps of the assay—steps which are numerous and exacting—which rely on a fixed or mobile antigens, enzyme-linked antibodies and enzymatic substrates to produce a subsequent reaction that is in some way detectable or observable. This typically involves several applications with reagents, detergents and a final washing step to remove all non-specific or unbound antibodies which is finally followed by an enzyme facilitated chemical conversion (with a substrate) which results in an absorbance or fluorescence in a specific range or spectrum identifying the adherence of a selective, tagged-antibody to a target molecule or antigen. Though exhibiting a well-documented and lengthy track record, assays are however in need of a laboratory, dedicated equipment and trained personnel, as seen above in PCR testing, but also necessitate binding and washing steps which require exacting, multi-step procedures in order to determine (1) presence or absence of desired target and/or (2) the overall quantify (e.g., concentrations) of said target. This lends more to a retrospective analysis in which antibodies are utilized to better detect the later stages of infection or past infection.
And while antibodies are a logically and naturally derived means of antigen (target) detection, responsible for identification and “tagging” of these antigens typically for a directed, immunological destruction of these targets through a series of actions including neutralization, agglutination, precipitation and complement activation, the antibody detection is only part of the process. To be observable, these antibodies must also be linked to an enzyme and exposed to a substrate specific to the enzyme wherein the resultant reaction (between enzyme and substrate) produces a detectable signal (e.g., a chromogenic change). As is the case with OTC pregnancy tests and rapid Strep tests, this method is particularly helpful to determine the presence or absence of a target but lacks sensitivity when a small number of targets are present. Equally, this method relies upon an appreciable color change for determinations which may indicate the existence of the target without informing the actual quantity of target molecule. What's more, relatively large amounts of substrate must be hydrolyzed by the bound enzyme before any appreciable detection can be achieved.
To overcome this infirmity, enzyme-linked fluorescence assays (ELFA) was introduced to improve sensitivity in samples exhibiting greatly lowered concentrations as compared to those substrates yielding strictly colored products (as above). Whereas ELISA is chromogenic, the substrate used in ELFA is fluorogenic and results in sensitivity rates which are on the order of 100 times more sensitive than ELISA. And, while greater sensitivity rates are achievable, the immunochromatographic process continues to rely on multi-step procedures (including the steps of procuring samples, fixing antibodies, antigen binding, and washing) and therefore requires many steps—both in preparation, actual analysis and reading of results.
And although a multi-step preparation and analysis is untenable in a rapid detection format, the use of fluorescence and chemiluminescence is of particular appeal and importance to inventors. Specifically, although these assay processes are highly labor and equipment intensive and require samples to be collected via blood collection and transported to laboratories in order to determine results, luminescence as a marker or indicator has seen use in the serological detection of disease for a number of years and bears a long track record in this regard. In fact, chemiluminescence or decay from an excited electronic state to a lower energy level resulting in an emitted light, has seen applications in gas analysis for poisons or other impurities, analysis of organic metabolisms, combustion analysis and biomolecule detection. This is especially interesting in the detection of biological markers (e.g., in forensic science) wherein the heme in blood may be utilized to detect even trace amounts of blood. Moreover, fluorescence has the unique ability to both qualify and quantify the amount of a target molecule which is over and above the capability of simple ELISA.
And while serological testing complements “rapid” viral detection, as an indication of current or past exposure, the development of pivotal ‘first-line’ methods of detection, especially in respiratory samples (seeing the highest concentrations during initial stages and peaking in the second week), that are truly “rapid” is an absolute imperative.
It is therefore chiefly critical to rapid detection that inventors have determined a means to provide improved detection methods, evidencing enhanced sensitivity and specificity, through a combination of (1) highly sensitive biological receptivity to a specific target molecule (biomarkers, compounds, pathogens (organic and inorganic), drug substances and poisons alike), whereby target binding produces near-instantaneous duplex formation and subsequent conformational changes resulting in a observable, single-step bio-illumination, in opposite of micro assays, that (2) can be optically detected without sequential (and multi-step) amplification, as in PCR, all via a non-invasive procedure. Moreover, it is equally advantageous for such a (1) biological binding and (2) optical detection unit to require little to no skilled technical intervention or reliance upon a laboratory (other than confirmatory results) which is completely contained within one device that is mechanically simple, largely reusable and capable of being reduced to a hand-held apparatus informed exclusively by utilizing exhaled breath.
While immunology (more singularly serology) and fluorescence may find their roots as far back as the 19th century, the combination of the two is only in its nascent stages of development when used for detecting and analyzing respiratory samples for content of specific target substances, molecules and compounds. In fact, breath gas analysis, or “breathprints”, have only seen development starting as late as 1971 when Linus Pauling first demonstrated that human respiration contains a series of gases and compounds that may be utilized to indirectly analyze, monitor and diagnosis patients for any number of biomarkers correlated with diabetes, lung cancer, head and neck cancer, asthma, COPD and cirrhosis. What is more, breath analysis itself has only recently become recognized institutionally as evidenced by the International Association of Breath Research (IABR) only first established in 2005 and having its first breath analysis summit in 2007 (First Breath Analysis Summit/3rd annual meeting of IABR in 2007).
Manifestly, while there have been attempts to detect compounds that are a byproduct or secondary indicator of disease, these endeavors have failed to move beyond secondary analysis and on to a solid correlation to directly observe and analyze causative agents non-invasively and in real time.
Thus, there is a significant, well recognized, and yet unmet need in the art for a means to detect a target molecule, compound or substance through nanomaterial-based technology wherein, for example, a target-specific, oligonucleotide hybridization probe (molecular beacon: MB) acts as an antibody alternative (i.e., an antibody mimetic/probe) which expresses a high affinity for a distinct target molecule, in the present case a viral infective agent. And although the present invention is a viral pathogen, molecular sequestration and visualization can easily be utilized for a compound, biomarker, bacterial pathogen, poison, chemo or biohazard or any other genetic sequence capable of target-specific binding and resultant molecular conformational change. Said MB is particularly useful in the detection and quantification of target molecules where said probe is capable of luminescence after binding, in a specific light wave range, which may be observed and analyzed optically via the methods disclosed herein.
It is the introduction of nucleic acid probes in the late 1990s, by the likes of Tyagi and Kramer, which provides just the alternative to antibody detection, and antibodies themselves, via an oligonucleotide hybridization probe, consisting of short DNA or RNA oligomer sequences, capable of effectively binding to a target molecule and providing a duplex formation with discrete conformational changes. The molecular beacon (MB) itself is a hairpin-shaped structure which is typically 20 to 40 nucleotides in length evidencing a “loop”, “stems” and fluorophore and quencher “end cap” portions. The “loop” structure is roughly 18-30 base pairs which complements the target molecule. Each “stem” portion attaches to the termini of the “loop” structure and are made of short (5 to 7 nucleotides) and reciprocal base pairs ensuring an essentially “closed” conformation. At the 5′ end of the molecular beacon strand is covalently bound a fluorescent dye (i.e., fluorophore or fluorochrome) and at the 3′ end exists a covalently bound quencher dye, which does not fluoresce. In this “closed” conformation, the fluorescence of the sensor is quenched due to the close proximity of the quencher to the fluorophore, ideally completely with very little residual fluorescence, through fluorescence resonance energy transfer (FRET) in the stem-closed formation wherein FRET is used to describe the transfer of energy between the fluorophore and the quencher in a non-radiative process, which is distance dependent. Concisely, the protein-induced conformational change in the molecular beacon resulted in a change of proximity between the fluorescence probes attached to the ends of the probe generating a fluorescence signal change from the “de-quenched” fluorescent dye.
In operation, once the complementary target nucleic acid sequence is “recognized” by the loop structure, the affinity of the loop nucleotides for the reciprocal target nucleotides supersedes the affinity of the base pairs in the stem, due in no small part to the larger ratio of loop structure nucleotides to stem nucleotides, thus forming a duplex with the target molecule. The molecular probe then transitions from an unstable “closed” configuration to the more stable “open” conformation wherein the fluorescent dye (fluorophore) and quencher dye are separated. A process called hybridization.
Therefore, in the absence of a target molecule, the higher stability of bonds between corresponding nucleotides in the stem greatly reduces the chances of a non-specific opening of the MB. Yet, with the introduction of a target molecule, and its recognizable nucleotide sequence, subsequent MB-target binding and hybridization occurs, the combined MB and target form a duplex, said duplex undergoes a conformational change thereby causing the fluorophore to separate from the quencher. The fluorescence signature thereby increases significantly due to diminished energy capture by the quencher caused by increased distance between fluorophore and quencher in the “stem-open” arrangement thus producing observable illumination in the presence of a specific target molecule.
The present invention seeks to address, mitigate and/or alleviate the above referenced infirmities and disadvantages of the current state in the art of target molecule detection and to allow for (1) direct observation of a target molecule, (2) through the binding of an illuminable molecular beacon and target compound, that (3) fluoresces, post binding, and is excitable to a specific light range (4) detectable via a specifically emitted wavelength, (5) observable via directed, filtered beams in an optical scanner—all in a (6) reusable, handheld device that gives (7) near-instantaneous results.
In the present invention's broadest terms, the disclosed device allows the rapid detection and measurement of target molecules in an exhaled breath condensate through target-specific “tagging” by a selective probe or beacon, subsequent collection of breath condensate containing said combined duplex of target and “tag” (e.g., probe) and detection and observation of the presence (and quantity) or absence of “tagged” molecules in the collected sample through enhanced light projection to a receiving detector. Succinctly, the present invention is dependent upon both target detection and a conformation-induced fluorescence which is selectively excitable to allow for target-specific recognition (qualification) and quantification through improved optical observation techniques. Such observation utilizes a non-energy dependent reflective, ellipsoid surface to “amplify”, direct and focus fluorescent light toward a sensor/detector for observation, data collection, analysis and conveyance of said data for medical diagnostics.
As well, inventors contemplate similar sensor beacons (e.g., quantum dot (QD) semiconductor particles) to be of particular interest whereby QD photoluminescence has greatly enhanced brightness, in the order of 20 to 100 times the brightness of a fluorescent dye, and high extension coefficient as a potential substitute for conventional donor fluorophores. Further QD size may have implications on coloration and the potential duration of fluorescence which may afford a greater variability between and among QDs as opposed to fluorescent dyes. Yet, although potentially promising, this technology bears some concerns as to safety and toxicity that may potentially adversely affect the test administrator thereby decreasing their utility.
Taking the foregoing into consideration, the term probe (i.e., molecular beacons, quantum dots, antibodies and antibody-like substances including mimetics or other organic or inorganic molecules capable of fluorescence or phosphorescing), in most cases, may be used interchangeably as performing the same basic function of detection and signaling, but as can be seen, each does so in very discrete and disparate ways. It is these differences which informs the present invention in terms of both form and function (i.e., unique operability).
The biological functions of target-probe conjugation and subsequent fluorescence, while integral to the functioning of the present invention provides, though, affords an incomplete analysis of the unique character of the present invention. In addition to the novel use of molecular beacons in non-invasive identification of a target molecule, in this case a specific virus, is further complemented with a new and novel means of biochemical detection. In opposite of a conventional spectrophotometer, where light (illumination) is provided to a sample and the sample emits light in an omnidirectionally and indiscriminately to a detector, the present means of detection directs, focuses and “pinpoints” light toward the detector (via an ellipsoid “concaving” mirror) which efficiently concentrates the detectable light which allows for conservative uses and the ability to detect even very small concentrations of fluorescence.
The present invention itself is thus comprised of (1) a biological component, (2) a collection reservoir and (3) an optical detection device (the later reservoir and device serving a dual purpose). And while each may be viewed independently, it is their relationship to one another, taken as a whole, that provides the combined novelty and utility to the present invention.
And while each one of these above advancements alone constitutes new and novel improvements individually, it is the combination of each of the biological, physical and optical processes and methods together which delivers a truly innovative means of rapid, repeatable and non-invasive target detection.
In a particular preferred embodiments, fluorescence or phosphorescence, can be induced through enzyme-linked bioassay, via conformational changes prompted by target-MB binding and resultant luminescence and or MB with the addition of quantum dots replacing the functional organic fluorophore. It is also within the contemplation of inventors that illumination, fluorescence or phosphorescence may be achieved through one of several means: chemiluminescence, a chemical reaction (the addition of the enzymes to a substrate containing reagent), electrochemiluminescence, radioluminescence (power derived a radiation source), electroluminescence, mechanoluminescence: a mechanical triggering event (ex. as a result of the physical binding of target to probe), piezoluminescence (ex. changing pressures), sonoluminescence (sonic pressure), thermoluminescence, singly, or by a combination thereof.
In addition to the MB-target, fluorescence and sensor device, the present invention apparatus, method of use and system employs one or more ellipsoidal mirrors in the focused collection of detection data. Ellipsoidal mirrors are currently used in various applications like focusing light into optical fiber, focusing lasers, reflective microscopy, spectroscopy, and focusing x-rays and are employed here to exponentially increase detection passively, through reflected light, resulting in an energy independent amplification thereby increasing target sensitivity requiring no additional resources for labeling, illuminating or identifying target compounds. Expressly, this application and use of ellipsoidal reflective properties is utilized to focus and concentrate small amounts of light emitted from a sample, to amplify and increase detection, qualify and quantify the emitted light and thereby allow the present system to identify and measure even minute amounts of target complexes directly. Advantages again exist here with MB-target duplexes over enzyme-linked immunoassay wherein fluorescence is autonomous bioluminescence resulting from conformational changes requiring no exogenous light source for excitation.
Too, the patient interface would consist of a disposable tube containing a sensor, into which the patient would expel breath at a certain rate and volume. Inventors contemplate a means to measure exhalation rates which may be consist of a manual peak flow meter, an auditory indicator (e.g., a whistle) or any automated device, manual or electrical, which determines force and/or length of exhalation.
In operation, if the patient's breath contains the desired target, said target would react with the sensor (i.e., the affinity of the MB, antibody or other antibody mimetic to a target would allow for target binding and hybridization). The tagged-target complex would undergo an illumination event (as described above). The amount of tagged-target present in the volume of breath can be directly “observed”, collected and recorded as related to the amount of light detected. This measure of complex illumination (i.e., measurable light units) may then be used to determine the quantity of the target substance whereby the amount may signify a ‘level’ that allows the observer to determine the presence of target substance, the amount of target substance, or both, in order to determine the effects of the presence and amount of said target substance.
The sensor device may consist of a PCR collection tube or tubes, or similar device(s), or and of a plurality of probes which may be coated with the specific MB or probe designed to bind with the target sample. It is in the further contemplation of inventors to have the interior surface of the collection chamber coated with target-specific probes wherein a specific target, if present, would bind to a probe, and the resulting complex may precipitate into a collection reservoir for detection and analysis. Ideally, the sensor is integrated into a hand-held device, and when incorporated into or inserted into the device, or the device is placed into a collection receptacle having same, the collection is suspended at one focal point of an ellipsoidal mirror.
The patient interface is disposable and may be designed specifically for a single substance or a plurality of substances, sequentially or contemporaneously. The optical detection device can be reused, sensors interchangeable and each can be designed for detecting a variety of substance molecules, light frequencies and wavelengths within the same device. The device may be designed to identify a specific patient interface and calibrate its ability to analyze a patient's breath accordingly (factoring variables, including but not limited to, sensor particle size, target size, complex weight, breath volume and the like).
While the novel features and method of use of the application are set forth above, the application itself, as well as a preferred method of use, and advantages thereof, will best be understood by referencing the following detailed description when read in conjunction with the accompanying drawings (in view of the appended claims), wherein:
And while the present invention, integrated system and method of use are amendable to modifications and alternative configurations, embodiments thereof have been shown, by way of example only, in the drawings and are described herein in adequate detail to teach those having skill in the art how to make and practice the same. It should, however, be understood that the above description and preferred embodiments disclosed, are not intended to limit the invention to the particular embodiment disclosed, but on the contrary, the invention disclosure is intended to cover all modifications, alternatives and equivalents falling within the spirit and scope of the invention as defined within the claim's broadest reasonable interpretation that is consistent with the specification.
The following description and accompanying drawings are illustrative and are not to be interpreted as limiting. Various key features are described to provide those having skill in the art the requisite description to made and use the same. However, it is to be understood, that certain features that are routine in the art are not described in detail that would prove pedantic to the skilled artisan. It bears including that the present invention may be practiced, in various cases, in one, some or all embodiments, preferred or simply described, with or without some of all details or features without departing from the spirit and scope of the invention as described.
References to “an embodiment”, “preferred embodiment”, “one embodiment”, “another embodiment” and “yet another embodiment” means that said “embodiments” are inclusive of all other “embodiments” in that a key feature, structure or iteration may be borne across all embodiments where their inclusion is not obviated by physical or structural inabilities to incorporate or combine said features, structures, or iterations or where their incorporation would not render the present invention inoperable for its intended use. To the contrary, “embodiments” as they are described herein are inclusive of all embodiments and not mutually exclusive of other embodiments. It is also in the understanding of inventors that not all features may be included in all embodiments while several features may be included in one embodiment or across embodiments.
Terms used within the present application are to be read as having their ordinary meanings in the art, as defined by the disclosure, and within the context of the surrounding defining language. Those terms, used by inventors, may as well be defined using specific terminology and descriptions are provided in the disclosure enumerating such terms and may have those meanings ascribed to each term as defined by those terms' accompanying language.
Section headings, titles and subtitles may be used strictly for organizational purposes and should not be interpreted as limiting the disclosed subject matter in any way. And while certain descriptions or embodiments may appear under a certain heading, title or subtitle, these descriptions and embodiments pertain to the invention and disclosure on the whole and should be read to apply to the entire description.
The present invention relates to the detection (qualification), analysis and quantification of a specific target molecule within an exhaled volume of breath, through and probe-reporter-detector model and process whereby occurs: (1) a high affinity attachment of said target to a fluorescent-tagged beacon (e.g., probe) to form a beacon-target complex, more technically a duplex, (2) conformational-induced, or enzyme-substrate facilitated fluorescence of said beacon or tag (making said duplex identifiable), (3) collection and/or concentration of said into a collection receptacle, (4) focusing endogenously or exogenously produced fluorescent light toward a detector which senses either the absence or presence of fluorescence and, if present, may use the overall measurable target-probe (via number or light intensity generated to quantify duplex populations through correlations with overall in vivo target concentrations.
As depicted in
In operation, once the complementary target 150 nucleic acid sequence is “recognized” by the loop 120, the affinity of the loop 120 nucleotides for the reciprocal target 150 nucleotides supersedes the affinity of the base pairs (A-T and C-G) in the stem 130, due to the number of loop 120 nucleotides in relation to stem 130 nucleotides, thus forming a duplex 180 with the target 150 molecule wherein the molecular beacon (MB) 100 is hybridized to the target 150. The MB-target duplex 180 then transitions from an unstable “closed” configuration 110 to the more stable “open” conformation 160 wherein the fluorescent dye (fluorophore) 122 and quencher 125 are separated due to hybridization.
Therefore, in the absence of a target molecule 150, the higher stability of nucleotide bonds between corresponding A-T and C-G nucleotides in the stem 130 greatly reduces the chances of a non-specific opening of the MB 100. Yet, with the introduction of a target molecule 150, and its recognizable nucleotide sequence, subsequent MB-target binding, duplex 180 formation and hybridization occurs, the combined MB 100 and target molecule 150 form as duplex 180, said duplex 180 undergoes a conformational change thereby causing the fluorophore dye 122 and quencher 125 to separate. The fluorescence signature (fluorescence signal 170) thereby increases significantly due to diminished energy capture by quencher 125 with co-temporal increased distance between fluorophore dye 122 and quencher 125 in the “stem-open” arrangement 160 thus producing observable illumination (via fluorescence signal 170) exclusively in the presence of a specific target 150 molecule.
In the present application, the target (MB 100), here a viral pathogen, is recognized by the probe (MB 100) wherein the target 150 (virus) and the probe 100 bind to form duplex 180. This binding is target-specific and only occurs in the presence of said virus target 150. In the absence of target 150, corresponding A-T and C-G nucleotides in the stem 130 would provide the most stable energy state and would remain in the “closed” conformation 110, providing no probe fluorescence (via fluorescence signal 170).
As provided in
The collection reservoir 220, which may take the form of a “PCR tube” or similarly translucent collection reservoir 220, is positioned at the most inferior portion, designated area B, or “trough” area of the present device 200, and opposite from area A and filter 290. It is contemplated that MB 100, or similar probes, may reside on the interior surface of chamber 250, within collection reservoir 220 or a combination thereof.
Completing the present device is the externally applied cooling system of
One preferred embodiment, as shown in
In operation, the device user places a rubber tube, mouthpiece, cover or like apparatus (not shown) at the opening of the 225 or like breath collection device, in patient's mouth and exhales, via a self-actuated air movement either normally or forcibly. The user's exhalation (ECB) collects on the collection chamber interior 250, accumulates, aggregates (i.e., precipitates) and “pools” into the collection reservoir 220—serving as a target sensor 520. The patient's exhalant continues in a vortex/cyclonic collector (see
In one preferred method, the sensor 520 contain MBs 100 that are specific for a particular target 150 (e.g., bacteria, virus, biomarker or inorganic marker). If the invention's user's breath (EBC) contains the desired target (i.e., pathogens or other sought particles or compounds) specific to the molecular beacon 100, the target 150 will bind to the molecular beacon 100 and form the duplex described above. The photon emission (as seen in fluorescence signal 170 of
As illustrated in
As depicted in
Equally in
From the above disclosure, many alterations and modifications, including preferred embodiments, of the present invention to a person having requisite skill in the art. Therefore, descriptions should be interpreted as merely illustrative and exemplary as providing the best mode or modes contemplated by inventors instructing the skilled artisan to make and use same which is susceptible to broad utility and application. Expressly features, structures, sizes, variations and configurations may be amended, restructured and reconfigured without departing from the scope and spirit of the present application.
The foregoing disclosure is not to be viewed in a limiting sense or to otherwise exclude any embodiments, adaptations or equivalent arrangements naturally following from disclosure of the present application. In opposite, the claims of present invention should be read giving the broadest interpretation to the inventive aspects of the invention described herein and only limited by the claims appended hereto including equivalents thereof.
U.S. Provisional Patent Application No. 63/021,054 filed May 6, 2020 PCT International Patent Application No. PCT/US2021/031196 filed May 6, 2021
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US21/31196 | 5/6/2021 | WO |
