Breath analyser and urea breath test method

Abstract

The present invention provides improved breath analyzers and breath testing methods to determine the presence of disease in humans, including but not limited to Helicobacter pylori in the digestive tract of a subject.

Description

[0001] Related Applications

[0002] This application claims priority to International PCT Application No. PCT / US2019 / 065550, filed December 10, 2019, and to U.S. Provisional Application No. 62 / 879,345, filed July 26, 2019, the entire contents of which are incorporated herein by reference. TECHNICAL FIELD

[0003] The present application generally relates to breath analyzers and breath testing methods for detecting gases in a human breath sample to determine whether a subject has a disease in their digestive tract. In some cases, the disease can include, but is not limited to, Helicobacter pylori. BACKGROUND

[0004] Helicobacter pylori (“H. Pylori”) affects approximately two-thirds of the world’s population, causing chronic gastritis and most peptic ulcers, and is a major cause of gastric cancer, the third leading cause of cancer death worldwide. Despite declining prevalence in Western countries since its discovery in 1982, H. Pylori remains at very high levels in Eastern Europe, Asia, and the Asia-Pacific region, affecting over 80% of young adults and children. In most cases, eradication of H. Pylori and prevention of gastric cancer is achieved through a combination of antibiotics and proton pump inhibitors (PPIs). An important property of H. Pylori is its abundance of urease, which hydrolyzes urea. When H. Pylori in the stomach is exposed to urea, it converts the urea to CO2and NH3, both of which reach the lungs via the bloodstream and are exhaled in breath. The breakdown of urea is described by the following equation:

[0005] CO(NH2)2+ H2O — urease — > CO2+ 2NH3

[0006] Detection of H. Pylori can be achieved through invasive and non-invasive diagnostic methods. Invasive methods include: 1) upper gastrointestinal endoscopy, biopsy for culture and rapid urease test (CLO test); 2) blood test for H. Pylori serum antibodies. Advantages of invasive methods are the accuracy of diagnosis by examining stomach tissue containing H. Pylori, and the accuracy of serum antibodies. Disadvantages are a) high cost, inconvenience, and risk of endoscopy; b) misdiagnosis due to failure to obtain tissue from the actual site of the stomach containing H. Pylori; c) serum antibodies for H. Pylori cannot confirm eradication (they persist for a long time regardless of whether the patient has H. Pylori).

[0007] Currently known non-invasive methods are 1) 13 C urea breath test 13The advantages of these methods are: a) the non-invasiveness of these methods, and b) the traditional 13 C UBT has high sensitivity (95%) and specificity (95%). 13 The disadvantages of C UBT are: a) the equipment is expensive (Otsuka America POCone infrared spectrometer, $25,000), and requires paid professional operation; b) 13 C) labeled urea is expensive; c) the breath test kit is relatively expensive ($110); and d) its availability at the point of care and directly to consumers as an over-the-counter product is very limited. Stool testing for H. pylori antigens is a) spotty; and b) has rather low sensitivity (85%) and specificity (85%). The shortcomings of both invasive and noninvasive diagnostic methods significantly limit the detection and subsequent eradication of H. pylori worldwide.

[0008] 13 C UBT is based on the hydrolysis of Helicobacter pylori catalyzed by its abundant urease 13 C-labeled urea ( 13 CO(NH2)2) and is produced during respiration 13 The capacities of CO2 and ammonia (NH3) are as follows:

[0009] 13 CO(NH2)2+H2O―Urease→ 13 CO2+2NH3

[0010] conventional 13 C UBT is provided as a kit containing 13 C-labeled urea powder (pranactin citrate drug powder), one bag for a baseline fasting breath sample and a second bag for ( 13 C-labeled urea) bags of postprandial breath samples. The bags are shipped to a centralized laboratory if they have the necessary equipment, or tested in the healthcare provider's office by trained personnel. This traditional 13 C UBT measures intake 13 C-labeled urea in the individual's breath 13 CO2 / 12 CO2 ratio. This is the increase in baseline fasting breath CO2. 13 C-labeled urea (rather than unlabeled urea) was used to demonstrate that 13 CO2 only comes from Helicobacter pylori 13The decomposition of C-labeled urea, rather than from other sources of CO2 release in the body (such as muscle). Although natural isotopes of carbon exist in nature, 13 C, but it only accounts for 1.8% of the total carbon. 13 The possibility that CO2 is a product other than urea hydrolysis by H. pylori is extremely remote.

[0011] above 13 The C UBT procedure is as follows: a) the individual abstains from antibiotics, PPIs, bismuth, and sucralfate for two weeks; b) on the day of the test, the individual fasts (no food orally) for one hour; c) the individual exhales 13 C) in bags provided by UBT suppliers; d) individual intake 13 C urea powder; e) 20 minutes after ingestion, the individual exhales into a bag provided by the vendor; f) a professional healthcare provider collects the bags and runs them through a spectrometer (if available on-site) or sends them to a centralized laboratory for testing; g) the individual is notified of the results within 48 hours of sending the bags to the centralized laboratory, or within minutes if a spectrometer is available on-site.

[0012] As described in the present disclosure, it is desirable to provide a handheld breath analyzer device that is easily operated by a non-professional; that will perform a novel urea breath test (UBT) by measuring NH3 and CO2 in a single breath sample (e.g., simultaneously); and that is inexpensive (e.g., $40-60 per device plus test). It is also desirable to provide such a device that will provide results immediately at the point of care and be available directly to consumers via over-the-counter (OTC) or online sales. Further, it is desirable to provide a device that can be used 13 C-labeled urea or inexpensive (e.g., less than $1 per test) unlabeled urea breath tests. Summary of the Invention

[0013] Certain embodiments of the present disclosure provide a breath analyzer. The breath analyzer includes an input, a first sensor, and a second sensor. The input is configured to receive a breath sample, and the first sensor and the second sensor are each configured to contact the breath sample. In some embodiments, the breath analyzer has two separate channels. In this case, the breath sample is configured to pass from the input through separate channels to each of the first sensor and the second sensor. The first sensor includes a first conductive polymer and a conductive material. The first conductive polymer contacts the conductive material and has a resistivity that increases in response to an increase in ammonia concentration. The second sensor includes a second conductive polymer and a conductive material. In some embodiments, the conductive material of the first sensor and / or the second sensor includes a plurality of electrodes. In some cases, the plurality of electrodes include wires arranged in a spiral configuration. In other embodiments, the plurality of electrodes include wires arranged in a rectangular configuration. The second conductive polymer contacts the conductive material and has a resistivity that increases in response to an increase in ammonia concentration. 12 CO2 and 13 The resistivity increases with CO2 concentration. The second conductive polymer comprises sulfonated polyaniline blended with polyethylene oxide. In some cases, the ratio of polyethylene oxide to sulfonated polyaniline is 10 wt% to 30 wt% (e.g., about 30 wt%). In certain embodiments, the sulfonated polyaniline is synthesized from a polyaniline polymer in the form of emeraldine. The breath analyzer also includes a processor and a circuit that operably connects the first sensor and the second sensor to the processor. The processor detects the resistivity in the circuit and uses the resistivity to calculate the total concentration of ammonia in the breath sample and 12 CO2 and 13 Total concentration of CO 2. In some embodiments, the breath analyzer communicates wirelessly with a remote device.

[0014] Other embodiments of the present disclosure provide a breath testing method comprising the following steps or providing a breath analyzer (which can be any breath analyzer described in the present disclosure, for example, including the breath analyzer described in the preceding paragraphs). The method further comprises prompting the subject to exhale a baseline breath sample into the breath analyzer, allowing the processor to measure the resistivity of the first sensor that occurs when the baseline breath sample contacts the first sensor, and allowing the processor to measure the resistivity of the second sensor that occurs when the baseline breath sample contacts the second sensor. The method further comprises providing the subject with a meal or capsule containing urea (wherein urea is 13C-labeled or unlabeled), then causing the subject to exhale a breath sample into a breath analyzer after urea ingestion. The method further includes allowing a processor to measure the resistivity of the first sensor that occurs when the breath sample after urea ingestion contacts the first sensor, allowing the processor to measure the resistivity of the second sensor that occurs when the breath sample after urea ingestion contacts the second sensor, and comparing the measured resistivity of the baseline breath sample to the measured resistivity of the breath sample after urea ingestion. The method further includes calculating the difference between the resistivity of the first sensor of the baseline breath sample and the measured resistivity of the first sensor of the breath sample after urea ingestion, and calculating the difference between the resistivity of the second sensor of the baseline breath sample and the measured resistivity of the second sensor of the breath sample after urea ingestion. Still further, the method includes expressing the difference in resistivity of the first sensor (e.g., in ppb of NH3) and the difference in resistivity of the second sensor (e.g., in ppm of CO2 or 13 CO2), and then displaying a final result as H. pylori positive when the difference between the post-urea and baseline measured resistivity of the first sensor and the second sensor is positive, and displaying a final result as H. pylori negative when the difference between the post-urea resistivity and the baseline resistivity of the first sensor and the second sensor is zero or negative.

[0015] Certain other embodiments of the present disclosure provide methods of detecting H. pylori in a digestive tract of a subject. The method includes collecting a baseline breath sample from the subject using a breath analyzer, and determining the total amount of ammonia present in the baseline breath sample and 12 CO2and 13 CO2using the breath analyzer. The method further includes allowing the subject to ingest a meal containing a predetermined amount of natural or labeled urea. The method additionally includes collecting a breath sample after urea ingestion from the subject using the breath analyzer, and determining the total amount of ammonia present in the breath sample after urea ingestion and 12 CO2and 13 CO2using the breath analyzer. The method further includes the step of indicating the presence of H. pylori in the digestive tract of the subject if the total amount of ammonia and 12 CO2and 13 CO2in the breath sample after urea ingestion exceeds the total amount of ammonia and 12 CO2and 13 CO2present in the baseline breath sample by a predetermined value. The breath analyzer used in the method can be any breath analyzer described in the present disclosure (e.g., including the breath analyzer described above, wherein the second conductive polymer comprises sulfonated polyaniline blended with poly(ethylene oxide)). BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1The acid-base (emeraldine salt (ES) - emeraldine base (EB)) transition of polyaniline is illustrated.

[0017] Figure 2 An embodiment of a PANI-CSA gas sensor is shown.

[0018] Figure 3 is a plot of sensor conductivity change illustrating one embodiment of an NH3 sensor.

[0019] Figure 4 is a plot of current change of varying conductivity illustrating a sensor based on Figure 3 NH3.

[0020] Figure 5 DropSens flow cell setup is shown.

[0021] Figure 6 is a calibration curve for CO2 showing the conductivity change of the NH3 sensor of the present disclosure based on the change in percentage of CO2.

[0022] Figure 7 is a plot showing the sensitivity of the PANI-CSA sensor of the present disclosure to specific gases.

[0023] Figure 8 A CO2 sensor coated with PPY-DBSA on a gold finger electrode is shown.

[0024] Figure 9 is a plot illustrating how the current of the PPY-DBSA sensor of the present disclosure changes when exposed to CO2.

[0025] Figure 10 Calibration of the PPY-DBSA sensor of the present disclosure is shown.

[0026] Figure 11 The effect of CO2, O2, H2O, and air on the PPY-DBSA sensor of the present disclosure is shown.

[0027] Figure 12 A benchtop prototype schematic of an embodiment of the breath analyzer of the present disclosure is shown, including a PPY-DBSA sensor and a PANI-CSA sensor.

[0028] Figure 13 A flow splitter valve for use in the multi-sensor system of the present disclosure is shown.

[0029] Figure 14 is a schematic of a flow splitter valve for use in the multi-sensor system of the present disclosure.

[0030] Figure 15A sensor contact pad of the present disclosure is shown connected to an analyzer by a clip and copper wire.

[0031] Figure 16 One embodiment of a portable breath analyzer of the present disclosure is shown.

[0032] Figure 17 An embodiment of a thin film interdigitated electrode for a gas sensor of the present disclosure is shown.

[0033] Figure 18 A manufacturing process for an interdigitated electrode of a sensor of the present disclosure is shown.

[0034] Figure 19a -c Various possible electrode layouts for a sensor of the present disclosure are shown.

[0035] Figure 20 PPY-DBSA sensors and PANI-CSA sensors according to certain embodiments of the present disclosure are shown.

[0036] Figure 21 The effect of 5% C02 on PPY sensors doped with different dopants (i.e., PPY-ASA, PPY-HBSA, and PPY-DBSA) is shown.

[0037] Figure 22 PPY-ASA sensors and PANI-CSA sensors according to certain embodiments of the present disclosure are shown, and the chemical structure of 3-aminobenzenesulfonic acid is also shown.

[0038] Figure 23 Stages of an electrochemical polymerization technique according to certain embodiments of the present disclosure are shown.

[0039] Figure 24 Another embodiment of a breath analyzer of the present disclosure is shown, including a PPY-ASA sensor and a PANI-DNNSA sensor.

[0040] Figure 25 A photograph of an electrochemical sensor prepared from a SPANI / PEO mixed solution.

[0041] Figure 26 A graph showing conductivity measurements of SPANI / PEO (30 wt%) when exposed to carbon dioxide.

[0042] Figure 27 A graph illustrating resistance changes of different conductive polymers to C02.

[0043] Figure 28 A graph showing C02 readings from a C02 meter sensor.

[0044] Figure 29 is a partially cutaway schematic perspective view of a prototype of a workbench of the present disclosure.

[0045] Figure 30 is a photograph showing a prototype of a workbench of the present disclosure.

[0046] Figure 31 is a graph showing the results of breath testing using a CO2 meter.

[0047] Figure 32 is a photograph showing two CO2 sensors, including a PANI / ABSA sensor and a PPY / ABSA sensor.

[0048] Figure 33 is a graph showing the CO2 response of SPANI with 10 wt% PEO.

[0049] Figure 34 A non-limiting embodiment of a wireless communication breath analyzer device of the present disclosure is shown. DETAILED DESCRIPTION

[0050] The following detailed description will read with reference to the drawings in which like elements in different drawings have like reference numerals. The drawings, which are not necessarily to scale, depict selected embodiments and are not intended to limit the scope of the application. Those skilled in the art will recognize that the examples provided herein have many useful alternatives that fall within the scope of the application.

[0051] Conductive polymers are widely used in gas sensor applications because they provide a stable and porous matrix for the gas components and also modify the electron transfer process. Recently, many efforts have aimed to improve the physical properties of conductive polymers (e.g. solubility, dispersibility, stability, and mechanical integrity). The formation of such composites is being optimized to produce materials that can be used in sensor manufacturing. The polyconjugated conductive polymer composite has recently been used in gas sensing applications because it has several useful properties, such as direct and easy deposition on sensor electrodes, thickness control, and redox conductivity of polyelectrolyte properties. These possibilities, together with the improvement of polyazole composites in terms of atmospheric stability, make them important candidates for specific technological applications. The advantages of conductive polymer composites for gas sensors are low cost, affinity to a variety of substrates, high sensitivity, and processability.

[0052] Although small amounts of NH3and CO2are produced in breath in conditions unrelated to H. pylori infection, in the case of ingestion of 13 C-labeled or unlabeled urea as a baseline pre-urea 13 NH3and CO2(and / or 13CO2) increases at the same time, ensuring that the source of these gases is only the effect of Helicobacter pylori on urea ( 13 As described herein, the co-inventors of this application have identified and optimized the hydrolysis of NH3, CO2 and 13 CO2 sensitive polymer-based sensor that can be docked on a device that will measure NH3 and CO2 (and / or 13 In many cases, these gases (NH3, CO2 and 13 The increase in CO2 can be detected no earlier than 10 minutes after ingestion and no later than 90 minutes after ingestion. Therefore, the individual should exhale into the present breath analyzer device, or the individual's breath sample should be taken before the individual ingests urea ( 13 C-labeled or unlabeled) substrate within 10-90 minutes (e.g., 20 minutes).

[0053] The present invention provides an improved breath analyzer and breath test method for determining the presence of Helicobacter pylori in a subject's digestive tract. Compared to existing methods and devices, the improved breath analyzer and breath test are less expensive, more convenient, and more accurate in diagnosis.

[0054] As described in more detail below, the innovative features of the breath analyzer (or breath analyzer device) of the present invention include, but are not limited to, the following: a) identifying and optimizing for simultaneous measurement of very low concentrations (such as those found in human breath) of NH3, CO2, and 13 The present invention also provides a polymer-based sensor for CO2; b) an interface for signals from two sensors when a gas sample passes through both sensors simultaneously; c) an interface for two independent sensors; d) software for simultaneously analyzing two or three different gas signals; and e) electronic circuitry and a display for the results. The display can show either digital or binary (e.g., positive-negative) results.

[0055] As described above, the novel breath measurement device of the present invention simultaneously measures NH3, CO2 and 13 The breathalyzer device consists of two sensors, one sensitive to NH3 and the other to CO2 and 13 CO2 sensitive. As used herein, CO2 refers to 12 CO2 (instead of 13 CO2). 12 CO2 and 13 The molecular weight of CO2 is very similar.

[0056] Although the term "simultaneously" is used throughout this disclosure to refer to the measurement of NH3, CO2, and 13The amount of CO2 can be measured, but those skilled in the art will understand that the measurements of these gases do not have to occur at exactly the same time. Instead, in any or all embodiments of the present disclosure, the breath sample can contact each sensor of the present disclosure simultaneously or approximately simultaneously. For example, when using the presently disclosed methods and apparatus, the breath sample will not travel to a first sensor and then to a second sensor. Instead, in certain embodiments of the present disclosure, the breath sample will travel to both sensors via equidistant (but separate) pathways from the mouthpiece to the sensors, such that the breath sample reaches both sensors simultaneously.

[0057] The urea breath test for detecting Helicobacter pylori is based on the ability of the bacterium (H. pylori) to convert urea into NH3 and CO2. In one embodiment, the urea breath test ("UBT") of the present disclosure utilizes unlabeled urea as an exogenous substrate for the H. pylori urease. This UBT method can measure NH3 in the desired range of 25 ppb–1 ppm and can measure CO2 (alone or in combination) in the range of, for example, 14,000-28,000 ppm or 20,000-50,000 ppm. 13 CO2 combination).

[0058] The UBT method and breath analyzer of the present disclosure are effective in certain individuals who ingest urea ( 13 C-labeled or unlabeled) and then detect CO2, 13 Increased levels of CO2 and NH3 gases. In particular, the CO2 sensor of the present invention is sensitive enough to detect 12 CO2 or 13 The breath test method includes the following steps: when the subject's breath sample shows an increase in CO2 concentration from a baseline of 100-1000 ppm. 13 Steps for diagnosing a subject as having Helicobacter pylori when CO2 or CO2 concentration increases relative to baseline. 13 None of the CO2 gases can be attributed solely to the hydrolysis of urea by Helicobacter pylori. For example, NH3 can be produced in kidney and metabolic diseases, while CO2 can be produced from muscle decomposition and colon microorganisms. However, the co-inventors of the present application found that the simultaneous increase in the two gases (NH3, CO2) shortly after ingestion of unlabeled urea (e.g., within 10-90 minutes after ingestion) ensures that the source of the increase in the two gases is the hydrolysis of urea by Helicobacter pylori. When urea is followed by NH3, CO2 and / or 13 CO2 values ​​are higher than the baseline NH3, CO2 and / or 13 When the percentage of CO2 is a certain predetermined percentage, UBT is positive for Helicobacter pylori infection. 13 CO2 values ​​are the same as or lower than baseline NH3, CO2 and / or 13CO2, the UBT is negative for H. pylori infection.

[0059] In one embodiment, the procedure for the UBT of the present disclosure is as follows: a) the individual will abstain from antibiotics, PPIs, bismuth and sucralfate for two weeks (as they can suppress H. pylori); b) the individual fasts (not by mouth) for one hour prior to the UBT; c) the individual exhales once into the input end (e.g., mouthpiece) of the breath analyzer device (no bag is needed to collect the breath), which is the baseline breath sample; d) the individual ingests a small solid or liquid meal (or capsule) containing 13 C-labeled or unlabeled (natural) urea; e) within 10-90 minutes after ingestion, the individual exhales again into the input end (e.g., mouthpiece) of the breath analyzer (again, no bag is needed), which is the post-urea breath sample; f) the results are calculated by the software of the device as the difference between the baseline values of NH3, CO2, and / or 13 CO2after urea ingestion (i.e., post-urea minus baseline), which is displayed on the display screen of the breath analyzer immediately after the second breath.

[0060] NH3sensor: Polyaniline (PANI) was first reported in 1862 and is one of the most studied and widely used conducting polymers. This is partly because it is a cheap monomer, but also because it is easy to synthesize in acidic aqueous solutions, environmentally stable, well processable, and soluble in common organic solvents (so it can be blended with other polymers).

[0061] Electrochemical polymerization of conducting polymers: The following five paragraphs relate to the electrochemical process that can be used to develop a PANI sensor (e.g., the PANI sensor of the present disclosure) or any electrochemical sensor. This process is shown in Figure 23 .

[0062] Typically, the experimental setup for the electrochemical synthesis of conducting polymers under laboratory conditions is relatively simple. In many cases, it involves a standard three-electrode electrochemical cell, although in some constant current polymerization cases a two-electrode cell can be used (Wallace et al., 2009). The polymer obtained by this procedure is deposited directly on the electrode. The anodic and cathodic electrolytes can pass through the electrode compartment at a specified flow rate, while the polymerization can be achieved at a constant potential. The most common experimental techniques used for the electrochemical polymerization of aniline are: cyclic voltammetry (potentiodynamic), chronopotentiometry, constant current, and constant potential.

[0063] Electrochemical polymerization using chronoamperometry is characterized by periodic regular changes in electrode potential, and throughout the experiment, the deposited polymer changes between its non-conductive and conductive (doped) states, then through a polymer exchange electrolyte (Heinze et al., 2010). At the end of polymerization, the resulting polymer is in its non-conductive form. Electrochemical oxidation of aniline monomer requires a high potential. Therefore, in the first 2-10 cycles, the upper limit of potential is high, but it can be lowered to avoid degradation due to over-oxidation of the pernigraniline form of polyaniline, due to the self-catalytic nature of aniline electropolymerization (Inzelt, 2008).

[0064] Electrochemical polymerization of aniline occurs together with the insertion of chloride anions (dopants) from the electrolyte, according to the following equation:

[0065] (PANI) n +nyCl - →[PANI y+ (Cl - )] n +nye -

[0066] Where y refers to the degree of doping, the ratio between the number of charges and the number of monomer units in the polymer (Kankare, 1998).

[0067] PANI exhibits three different oxidation states: leucoemeraldine (LEB, fully reduced), emeraldine (EB, semi-oxidized), and pernigraniline (PNB, fully oxidized). However, the protonated form of EB, Emeraldine salt, is the only conductive form, usually obtained by protonating the basic amine and imine sites in EB with strong acids. This is an important property of PANI, especially for ammonia detection, as it deprotonates the amine group in the emeraldine salt, converting it to the emeraldine base form, with a corresponding decrease in conductivity by several orders of magnitude. This transformation of polyaniline (from emeraldine salt (ES) to emeraldine base (EB)) is shown in Figure 1 The reaction that allows this change in conductivity is as follows:

[0068]

[0069] PANI doped with camphorsulfonic acid (CSA) has been shown to have a pH sensitivity of about 70 mV, which is higher than the 59 mV typically observed with other small counterions such as Cl - and SO4 2- . This is the basis for choosing CSA over other protonic acids for the PANI ammonia sensor.

[0070] PANI has the characteristics of chain flexibility, strong redoping ability, high dispersibility in many organic or aqueous solvents (depending on the dopant). In particular, sulfonated polyaniline, explained in more detail below, has high sensitivity to carbon dioxide, high stability and excellent electrical conductivity.

[0071] Manufacture of NH3 sensor: Non-limiting embodiments of the NH3 sensor of the present disclosure are shown in Figure 2 To manufacture such a sensor, the following steps were performed. HCI-doped PANI was prepared by chemical oxidative polymerization of aniline in an acidic aqueous solution (1 M HCI) with ammonium persulfate (APS) as the oxidizing agent. It has been previously reported that a higher polymerization yield can be obtained by using a ratio of oxidizing agent to monomer of 1.2. The mixture was left to polymerize overnight at room temperature. The PANI precipitate was collected on filter paper and repeatedly washed with 0.1 M HCI, then repeatedly washed with acetone. De-protonation of the resulting PANI salt was performed by stirring the powder in 0.1 M aqueous NH4OH at room temperature for 24 hours to obtain emeraldine base (EB), which was then repeatedly washed with water until neutral pH, then vacuumed at 60 °C for 48 hours. The PANI was re-doped with CSA in a molar ratio of 2:1. Thin film interdigitated platinum electrodes (IDA) (line spacing 100 pm) were purchased from the Center for Electronics Design and Engineering at Case Western Reserve University. Thin films were prepared by spin-coating a PANI-CSA solution on the IDA electrodes. Prior to spin-coating, the electrodes were cleaned by rinsing in methanol then deionized water and dried in a stream of dry nitrogen. The PANI thin films were then spin-coated onto the IDA Figure 2 ).

[0072] NH3 sensor properties: Calibration Figure 3 and Figure 4 25 ppm ammonia was first continuously diluted to 2.5 ppm, 250 ppb and 25 ppb, i.e. 10-fold dilutions in tedlar bags, respectively. Samples were collected from the tedlar bags using a 20 cc syringe, which was then directly injected onto the sensor. DropSens flow cell setup Figure 5 ) was used to inject ammonia gas onto the sensor, which was placed in the center of the DropSens flow cell. Teflon tubing was used to connect the gas to the flow device to minimize any NH3 absorption.

[0073] Measurements were performed by applying a fixed potential of 1 V to the sensor (contact pads) and measuring the resulting current, which varied with the gas passing over the surface of the sensor Figure 3 and Figure 4). The measurements were performed using an Agilent 4155C semiconductor analyzer and custom test software was developed using the EasyDesktop software that comes with the analyzer. Similar changes in conductance and current were observed when using a 1010-type gas diluter. A CH Instrument 660D-type potentiostat analyzer (CH Instrument, Austin, TX) was used to measure the sensor resistivity. The observed changes in resistivity at 250 ppb and 25 ppb were approximately 4.8% and 2.5%, respectively, which are high enough to reliably analyze breath samples at trace level ppb values.

[0074] Sensitivity of NH3 sensor of the present invention: Human breath contains trace amounts of C02, N2, 02, NH3, NO, H20, methane, volatile organic compounds (VOCs), and the like gases. To evaluate the effect of breath C02 at 4-5% concentration on the PANI-CSA sensor, the co-inventors of the present application tested a range of C02 concentrations. As shown in Figure 6 , the effect at 4-5% C02 concentration was negligible and thus unmeasurable. The overall sensitivity of the PANI-CSA sensor to NH3, C02, N2, and atmospheric air is shown in Figure 7 . The PANI-CSA shows a much higher response to NH3 compared to C02, N2, and air, considering that the concentration of NH3 is 25 ppm (0.000025%) while 100% dry gas is used for C02 and N2 (human breath contains 4-5% C02 and trace amounts of N2).

[0075] 13 C is a natural isotope of carbon, present in nature at 1.8% of total carbon. Thus, the increase in 13 C02 concentration is highly unlikely to be a product other than the hydrolysis of H. pylori urease 13 C-labeled urea. The polymer-based C02 sensor of the present disclosure is also sensitive to 13 C02 and can detect C02 and 13 C02 individually or cumulatively. As a result, the present invention can be used to detect H. pylori after ingestion of 13 C-labeled or unlabeled urea.

[0076] C02 sensor: Most of the work reported on C02 gas sensing uses doped or undoped Sn02. There is less attention to conducting polymers such as polypyrrole, which has unique properties for C02 gas sensing, such as low density, versatility of production methods, high anisotropy of conductivity, and non-metallic temperature dependence of electrical conductivity. Polypyrrole (PPY) can be prepared by a variety of methods such as chemical, electrochemical, and gas phase routes.

[0077] It has been reported that when polypyrrole, an organic polymer, is oxidized, it becomes a conductive polymer. The present co-inventors have tested the response of doped polypyrrole to 5% CO2 Figure 21 The CO2effect was more pronounced when polypyrrole was doped with 3- aminobenzenesulfonic acid (ASA) than when polypyrrole was doped with 4- hydroxybenzenesulfonic acid (HBSA) or 4-dodecylbenzenesulfonic acid (DBSA), and thus the response of PPY-ASA to CO2was significantly higher than that of PPY-DBSA or PPY-HBSA.

[0078] CO2sensor fabrication: In one embodiment of the disclosure, a CO2sensor, in particular a PPY-based sensor, was fabricated using dodecylbenzenesulfonic acid (DBSA) as a dopant Figure 8 Pyrrole (Aldrich) was dried with CaH2for 24 hours and then distilled under reduced pressure. Ammonium persulfate (APS, Kanto Chemical Co. Inc.) was used as an oxidant and dodecylbenzenesulfonic acid (DBSA, Aldrich) was used as a dopant. Under vigorous stirring, 0.15 moles of DBSA and 0.3 moles of pyrrole were dissolved in 500 ml of distilled water. 0.06 moles of APS in 100 ml of distilled water was slowly added to the above solution maintained at a temperature of 0°C. The obtained PPY powder (1 g) was completely dissolved in 25 ml of m-cresol or chloroform only by ultrasonication, 1 g of DBSA was added, and filtered through a 1 pm Teflon membrane filter. The solution was transferred onto a gold circuit interdigitated platinum finger electrode (Case Western University), the solvent was dried, and a high quality self-supporting thin film with a thickness of about 100 pm was obtained. FT-IR was used to characterize the characteristic peaks of the dopant and pyrrole. The smooth and coherent thin film obtained by the above preparation was cast onto another wafer surface for examination under a scanning electron microscope (SEM).

[0079] CO2sensor resistivity: DropSens flow cells were set up Figure 5 ) to examine the properties of the PPY-DBSA sensor of the disclosure. The current (resistivity) change of the PPY-DBSA sensor when exposed to 5% CO2 Figure 9 ) was measured using a 660D CH Instrument potentiostat analyzer (CH Instrument, Austin, TX) using amperometric i-t technique. The sensitivity of the PPY-DBSA sensor to CO2, O2, H2O, and atmospheric air was tested.

[0080] CO2 Sensor Calibration: The PPY-DBSA sensor of the present application was calibrated using 100% dry gas. A Custom Sensor Solutions Model 1010 precision gas diluter was used to deliver the analyte gas (CO2) to the sensor in the center of the DropSens flow cell. Mass and volume flow meters (Omega Engineering, Inc., Norwalk, CT) with a precision of ±0.8% were connected to record the flow. The change in resistivity was measured as a function of CO2 concentration Figure 10 ). The CO2 sensor was tested for its response to O2, N2, H2O, and air Figure 11 ). The effect of air was negligible and the effect of 100% O2 gas was minimal. However, H2O had a greater response on the resistivity of the CO2 sensor.

[0081] Both sensors of the present application (CO2 and NH3) need to be optimized, as described in more detail below.

[0082] NH3 Sensor Optimization: The co-inventors of the present application developed a device to make PANI-CSA sensors and examined their properties. A published report showed that the baseline concentration of NH3 in the breath of patients positive for H. pylori infection ranged from 0.02-0.17 ppm and increased to a range of 0.05-1 ppm in the breath after ingestion of unlabeled urea.

[0083] Effect of CO2, N2, O2, and H2O: To further optimize the PANI-CSA sensor, the PANI-CSA sensor was exposed to conditions similar to those in human breath. Human breath contains mainly CO2, N2, O2, and H2O. According to preliminary data from the co-inventors, the sensor had minimal response to 100% CO2 and 100% N2 and negligible response to air. Since the CO2 and N2 contained in human breath are far below 100%, the interference of CO2 and N2 in breath is negligible and thus insignificant.

[0084] Effect of humidity on NH3 sensor: NH3 is hydrophilic and the humidity of human breath can interfere with the sensor’s response to NH3. To address the effect of humidity, a desiccant was used and titrated using a VTI RH-200 relative humidity generator.

[0085] CO2 Sensor Optimization: The co-inventors of the present application have developed a device to make PPY-DBSA sensors that are highly sensitive to CO2. According to a published report on the gold standard 13 C UBT, patients infected with H. pylori had a concentration of 13 C labeled urea in their breath after ingestion 13CO2 is elevated, and in breath 13 The percent increase in CO2 over the CO2 baseline is between 2.4% and 4.4%, depending on the amount of urea ingested. Based on these findings, the co-inventors of the present application believe that the percent increase in CO2 over baseline after ingestion of 150-300 mg of unlabeled urea will be similar to that after ingestion of 13 C-labeled urea. 13 CO2.

[0086] Based on sensor calibration Figure 10 ), it is believed that the CO2 sensor of the present disclosure will be sensitive even at the lowest percent increase in CO2 in patients infected with H. pylori. Although there is no report of PPY-DBSA being sensitive to NH3, the co-inventors of the present application will check if the sensor is sensitive to NH3, and if so, will place a NH3-specific filter (e.g., 3A formed crystalline metal aluminosilicate) directly over the CO2 sensor to prevent NH3 from contacting the CO2 sensor. This filter is a desiccant that blocks NH3 and H2O.

[0087] Effect of CO2, O2, H2O, air, and NH3 on CO2 sensor: The co-inventors of the present application have tested the response of an embodiment of the CO2 sensor of the present disclosure to CO2, O2, H2O, and air Figure 11 ). Since the effect of air is negligible, it is believed that the effect of O2 and N2 in human breath on the CO2 sensor is negligible, since the O2 and N2 content in human breath is much lower than the 100% gas tested by the co-inventors. It is believed that there is no need to further check the effect of O2 and N2 on the CO2 sensor.

[0088] Effect of humidity on CO2 sensor: Preliminary data by the co-inventors indicate that humidity affects the resistivity of the PPY-DBSA CO2 sensor Figure 11 ). To counteract this effect, a desiccant can be used, and the amount of desiccant can be titrated by using different amounts and checking their effect on the resistivity of the CO2 sensor in the presence of the desiccant. A VTI RH-200 relative humidity generator can be used to generate various levels of relative humidity (RH), and calculate the amount of desiccant needed to achieve optimal sensitivity (resistivity change) to CO2 and / or 13 CO2.

[0089] Figure 15 An embodiment of the portable handheld breath analyzer device of the present application is shown. As described above, this device measures NH3 and CO2 (and / or 13CO2) to detect H. pylori infection. In the benchtop prototype of the present device, the co-inventors of the present application will connect two sensors (e.g., PANI-CSA and PPY-DBSA or SPANI); will standardize the prototype for two gases (CO2and NH3); and will optimize the prototype for final use with breath samples from H. pylori patients.

[0090] Non-limiting embodiment of the benchtop prototype: In one embodiment of the present disclosure, using the manufactured and optimized PANI-CSA and PPY-DBSA sensors, a benchtop prototype can be established as shown in Figure 12 DropSens 1 for PPY-DBSA sensor to measure CO2, DropSens 2 for PANI-CSA sensor to measure NH3. In another embodiment of the benchtop prototype, the DropSens flow cells are removed and the configuration of the benchtop prototype will have no flow cells. In this embodiment, the sensors will be connected directly to the CH instrument for gas analysis.

[0091] The two DropSens flow cells are connected to a two-way gas flow valve 5 by pipes 1, 2, 3, 4. A photograph and schematic of the two-way valve are shown in Figure 13 and Figure 14 A humidity filter and / or desiccant will be placed in pipes 1 and 2 in the configuration (circular, spiral, etc.) that optimizes the flow of gas mixture to each DropSens. Pressure gauges will be installed on pipes 1 and 2. The pressure gauges will adjust the flow of gas (or breath) in each pipe (1, 2) so that the flow of gas to each sensor (DropSens 1 and DropSens 2) is the same in each pipe with the desiccant present. The skilled artisan will appreciate that the desiccant can alter the flow of gas through the pipes, which in turn can be adjusted by using the pressure gauges in pipes 1, 2.

[0092] A two-way gas flow valve will be used to transfer the gas mixture to each side of the device during optimization of the benchtop prototype. The analysis gas is pre-processed to remove water, particles in the air, or unwanted gases, and then characterized by each gas sensor. In the case of two gas sensors, such as the benchtop prototype of Figure 12 The main gas flow will be divided into two separate gas flows so that they can be pre-processed by a flow splitter valve (e.g., as shown in Figure 12 and Figure 13 The ratio of the flow of the two outlet streams can be adjusted by opening one or both pressure gauges.

[0093] Turning to one or the other sensor is only for individual characterization and calibration of each sensor. Ultimately, the mixed gas containing NH3 and CO2 will enter both sensors, and the valve will be open to both directions. Each sensor will receive the same gas mixture, as is the case when an individual breathes into the device. Then, the same breath will enter each sensor. This explains why the NH3 sensor is made so that CO2 has no effect on it, and why the CO2 sensor is made (and a desiccant that will block NH3 is placed directly above it) so that NH3 has no effect on it. The co-inventors of the present invention have demonstrated from the experiments described herein that the rest of the elements of the breath (other than CO2 and NH3) have no effect on either sensor.

[0094] Measurement of NH3 and CO2: In the proposed bench setup Figure 12 ), the sensors will be exposed to a gas mixture (NH3, CO2 and / or 13 CO2) as described above. The contact pads Figure 2 , Figure 5 ) of the sensors will be connected to the analyzer Figure 15 ) through a clip and copper wire. The current co-inventors will use an Agilent 4155C semiconductor analyzer and custom test software developed using the EasyDesktop software provided with the analyzer. The measurements of the two or more gases NH3 and CO2 (and / or 13 CO2) will be obtained by applying a fixed potential of 1 V on the sensor for 200 seconds and measuring the resulting current change as a function of the gas passing through the sensor surface.

[0095] Method to improve CO2 (and 13 CO2) affinity of CO2 sensor:

[0096] Method 1 to make CO2 sensor: amine modification of PPY

[0097] Amine modification of sorbents is often used to introduce CO2-philic functionality. In principle, the nitrile groups in PPY can be reduced to primary amines using a variety of reagents. However, reactions that work well with low molar mass compounds are not always effective when applied to high molar mass polymers.

[0098] Polymers based on polypyrrole (PPY) have been used for CO2 detection because the aromatic amine is the focus of specific chemical interactions. Although PPY exhibits high electrical conductivity and sensitivity to CO2, it is difficult to use (e.g., due to its rigid backbone and low solubility) and has poor stability. This is primarily because the aromatic five-membered rings are covalently linked to each other, and in particular in the doped state, there is very limited rotational freedom, poor solubility. Polythiophenes are also five-membered rings and have similar issues. In the 1990s, others tried to solve this problem by alkylating thiophene monomers. These polymers became soluble, even processable, in the undoped state and caused widespread interest. However, upon doping, the rigidity of each aromatic unit limits solubility, regardless of the alkyl group. The skilled artisan will appreciate that chemical modification is a complex synthesis process and dispersants make the process less precise.

[0099] To increase the amine content and thereby improve the CO2 (and 13 CO2) affinity of embodiments of CO2 sensors, the present co-inventors used diaminonaphthalene sulfonic acid (DANSA) as a dopant. The addition of the primary amine enhanced the CO2 affinity compared to the parent polymer, as evidenced by the present co-inventors' gas adsorption experiments on film-like sample coatings on electrodes and gas characterization studies of membrane sensors.

[0100] Another method of making CO2 sensors is to polymerize polyaniline and oxidize it with 3-aminobenzenesulfonic acid. PANI is a more stable polymer than polypyrrole (PPY), and the sensor produced by doping PANI with 3-aminobenzenesulfonic acid (ASA) has a much larger response to CO2 than a PPY-ASA sensor.

[0101] Method 2 for making CO2 sensors using sulfonated polyaniline (SPANI)

[0102] An ideal polyaniline is sulfonated polyaniline ("SPANI"), which is self-doping due to the covalent attachment of sulfonate groups to the polyaniline backbone. SPANI is also water-soluble, as the additional sulfonate groups provide water solubility (which is suitable for gas sensing but can be problematic in aqueous applications). Alternatively, water-soluble poly[bis(4-phosphono)phenoxy]phosphazene (PPAP) is a good candidate as a dopant for PANI. Freshly prepared and dried acid-functionalized aryloxypolyphosphazene (PPAP) is dissolved in 10 mL of water at 0–5°C in a round-bottom 100 mL flask, and aniline monomer (ANI:PPAP ratio of 2:1) is added with vigorous stirring. Then, aqueous ammonium persulfate (aniline / oxidant ratio of 1:1) is slowly added to the reaction mixture as the oxidant. The reaction turns green within 1 hour and is stirred overnight. The dark green solution obtained by dialysis membrane was transferred to a polytetrafluoroethylene petri dish and dried in a vacuum oven at 50 °C overnight to obtain SPANI, which was confirmed by FTIR and P-NMR.

[0103] Method 3 for fabricating CO2 sensors using water-soluble sulfonated polyaniline (SPANI)

[0104] For SPANI synthesis, emeraldine hydrochloride PANI polymer powder (1 g) was dispersed in 150 ml of dichloroethane (DCE) at 80°C. Chlorosulfonic acid (1.2 ml) diluted with 2 equivalents of 15 ml of DCE solution (i.e., 15 ml + 15 ml) was prepared, added to the dispersed solution for 30 minutes, and stirred for 5 hours. If the first 15 ml was used up, the second 15 ml could be used until the total reaction time was 30 minutes. The solid product was then collected by filtration, carefully washed twice with cold water, immersed in 200 ml of distilled water, and heated at 100°C for 4 hours to hydrolyze it. After hydrolysis, the green solution was evaporated to dryness in a vacuum at 40°C. The crude SPANI powder was collected, washed with methanol to remove impurities, then filtered and dried in an oven at 40°C.

[0105] Alternatively, an aqueous SPANI solution can be prepared by dissolving SPANI in water. The structure can be confirmed by P-NMR, and the concentration of SPANI in the aqueous solution can be measured by UV-visible spectrophotometry. Other solvents (i.e., other than DCE and water) can also be used to dissolve SPANI, as long as the solvent can dissolve PANI and the acid.

[0106] Sulfonated polyaniline (especially when prepared by method 3 above) shows good solubility in water. However, the low evaporation and hydrophobicity of the finger electrodes are the result of poor adhesion to the electrodes and the polymer. To address these problems and increase the adhesion of the film, polyethylene glycol ("PEG") or polyethylene oxide ("PEO") are used due to their low glass transition temperature (T g) and hydrophilicity and is used as an adhesive.

[0107] Alternative methods to increase film adhesion may include the use of electrochemical polymerization or PPY / ABSA films (4-acetamidobenzenesulfonyl azide) (ie, polypyrrole doped with ABSA). Figure 32 Two example CO2 sensors are shown in FIG, including a PPY / ABSA sensor and a PANI / ABSA sensor (ie, polyaniline doped with ABSA). Applicants found that the conductivity of the PANI / ABSA sensor was low (~1E-12).

[0108] like Figure 25 As shown, three different mass ratios of PEO (10 wt%, 20 wt%, and 30 wt%) were mixed with SPANI and drop-cast onto a finger electrode. However, with the low PEO ratio (10 wt%), almost no phase separation was observed. However, with the high PEO ratio (30 wt%), the film had a homogeneous single-phase appearance. The baseline conductivity in air was 60 ± 5 μA at 0.5 V.

[0109] Conductivity measurements are performed with a CH instrument by applying 0.5 V and measuring the resulting current. Figure 26 As shown, the baseline (air) shows stable values ​​and low noise. The current change due to 5% CO2 is shown, and the baseline can be restored in a very short time when purging with air.

[0110] Four different conductive polymer electrochemical sensors were prepared and tested for their response to CO2. The conductive polymers tested included PPY / ASA; PPY / HBSA; PPY / DBSA; and SPANI. The test gas was 5% CO2. When the SPANI sensor was tested, the resistance change after exposure to CO2 was 40%. These results are shown in Figure 27 It was demonstrated that SPANI has high potential for CO2 sensing applications.

[0111] Figure 28 This is a graph showing the CO2 readings from the CO2 meter sensor. The CO2 meter has an operating range below 5% and a sensitivity around 100ppm.

[0112] Figure 29 and Figure 30 The workbench prototype is shown. Figure 29 As schematically illustrated in FIG, the prototype has dual channels for CO2 and NH3 gas characterization. Specifically, the workstation includes an NH3 sensor, an NH3 channel, a CO2 channel, a CO2 meter, and a temperature and humidity sensor. The ammonia and CO2 channels are configured to receive breath samples through a tube (not depicted) that serves as a mouthpiece. Figure 30NH3 sensor position indicates the space in which the ammonia sensor was placed in the prototype. The skilled artisan will appreciate that the location of the ammonia sensor can be different than that shown in Figure 30 The CO2 meter for the bench prototype was an industrial CO2 sensor (ExplorIR-M 5% CO2 sensor) used to study the prototype device.

[0113] The bench prototype also included a temperature and humidity sensor, one possible location of which is shown in Figure 30 Most of the time, the temperature and humidity sensor will indicate a room temperature (e.g., about 70 degrees Fahrenheit). The humidity for the NH3 sensor and the CO2 sensor is expected to be different. This is because, in most cases, the humidity of the breath will be prevented from being around 80-90% for the NH3 sensor (e.g., by using a desiccant). In some embodiments, a desiccant will be used in the CO2 channel to prevent the humidity of the breath sample to some extent before it reaches the CO2 sensor.

[0114] Figure 31 is a graph showing the results of breath testing using a CO2 meter. As shown, the signal of the CO2 meter becomes saturated with breath, with a working range of 0-5% CO2.

[0115] Figure 33 is a graph showing the CO2 response of SPANI with 10 wt% PEO. As shown, using the SPANI-PEO (10%) sensor, the noise is reduced a lot (compared to PANI / ABSA). The current change at 1% CO2 (I%) is 13%.

[0116] Conductivity measurements and closely related salinity measurements are important in many applications of gas characterization and sensing. Figure 17 A typical layout of interdigitated electrodes in the field of gas sensors is shown. Extensive research has been done on H2, O2, NO x , CO, etc. In addition, materials and circuit layouts have been found to improve the performance of these special gas applications. Various metallic materials, including gold, silver, platinum, etc., deposited on top of a dielectric substrate can provide the required working potential and interfacial properties.

[0117] Conductive materials are coated on the electrodes of the present invention to complete the current circuit for characterization of electron flow. Conducting polymers, such as polyaniline, are the interaction films for the present gas sensors. Attachment of gas molecules with different affinities changes the electrical properties of the thin films and are reflected in the current measurements. The electrode layout has also been reported to affect the performance of the current characterization. Many alternative layouts are shown in the figure, which are prepared for the gas analyzer device.

[0118] Manufacture of electrodes for gas characterisation: A planar micro-supercapacitor configuration can be chosen where the interdigitated electrodes are fabricated on the same plane and isolated by physical separation. This configuration has the advantage of ease of fabrication and flexibility due to the choice of electrodes. Figure 18 A non-limiting example of the manufacturing process of the sensor of the present device is shown. The patterned electrodes are fabricated through three main steps: photoresist deposition, patterning and thermal decomposition. A 5 mm thick alumina nitrile substrate is used to electrically isolate the supercapacitor from the substrate. SU-8 photoresist (MicroChem) is spin-coated on the substrate to a thickness of approximately 10-100 pm. After a soft bake at 115 °C for 5 minutes, the interdigitated electrodes are patterned by UV lithography. After exposure to Ar (argon) plasma PVD treatment of gold, silver or platinum, the chip is transported to a sputter coater. The current collector is then deposited using evaporation of 100 nm Ti / 400 nm Au layers, annealed at 250 °C for 20 minutes and patterned to remove the photoresist and obstructed metal using a conventional lift-off process.

[0119] The electrodes used in any of the sensors of the present disclosure can comprise thick film printed interdigitated platinum, gold or silver lines on a 0.6 mm thick alumina substrate, which provides excellent adhesion to the printed lines. Such electrodes are used by spreading the material over the interdigitations. The pads allow contact wires to be made for electrical connection.

[0120] Electrode layout: The circuit layout determines the performance and consistency of the conductivity measurements. Several circuit layouts for interdigitated electrodes are described below and shown in Figure 19a-19c Any of these electrode layouts can be used in any (or all) of the sensors of the present invention.

[0121] The electrodes can have a single channel or a multi-channel arrangement. One possible electrode design is a finger electrode as shown in Figure 19a The gap between the electrodes determines the total current flow, which is required to be 1 pA. Another electrode layout (as shown in Figure 19b comprises two elongated wires formed in a rectangular configuration. Yet another electrode layout (as shown in Figure 19c comprises two wires that are extended into a spiral structure. Alternative electrode configurations are also contemplated and are within the scope of the present disclosure.

[0122] In another embodiment of the present invention, the NH3and CO2or 13 CO2sensors are each placed in their own enclosed chamber within the device. In this embodiment, the breath sample is introduced into the device through a mouthpiece, and each sensor receives a portion of the same breath sample. The enclosed chamber of each sensor allows for the placement of a filter directly adjacent to each sensor to block specific gases and / or desiccants without affecting the other sensors’ measurement of the same breath content of NH3, CO2or13 Response to CO2.

[0123] In some embodiments, the breath analyzer device can connect to and communicate with a remote device (e.g. a smartphone, wireless hub, computer, or server in the cloud) using one or more communication technologies including, but not limited to, wired communications such as USB, wireless and cellular communications such as WiFi, Bluetooth, ZigBee, GSM, LTE, and infrared. Data can be transmitted in plain text or encrypted.

[0124] Figure 34 A non-limiting embodiment of a wireless communication breath analyzer device is shown. An exhaled breath sample is exhaled into the mouthpiece of the breath analyzer device and processed by the breath analyzer. Results are then archived and sent to a computer, one or more smart devices, or the cloud.

[0125] In some embodiments of the breath analyzer device, data transmitted from the breath analyzer device includes data from direct measurements of gas concentrations, as well as device health information such as device temperature and battery status. Data transmitted from the breath analyzer device can be raw data read directly from sensors, or can be data processed at the breath analyzer device after signal conditioning. Transmitted data can also be derived data such as patient health status, status of a disease from which the patient is suffering, or health data.

[0126] In some embodiments of the breath analyzer device, data transmitted to the breath analyzer device from a smartphone, wireless hub, computer, or server in the cloud can include device software updates, device parameter updates, and relevant information about the user or the user’s health, well-being, or disease status.

[0127] While certain preferred embodiments of the present application have been described, it is understood that various changes, modifications and alterations can be made therein without departing from the spirit and scope of the application as disclosed in the accompanying claims.

Claims

1. Breathalyzer, including: receiving an input of a breath sample; a first sensor in contact with the breath sample, wherein the first sensor comprises a first conductive polymer and a conductive material, wherein the first conductive polymer contacts the conductive material, wherein the first conductive polymer has a resistivity that decreases in response to an increase in ammonia concentration; a second sensor contacting the breath sample, wherein the second sensor comprises a second conductive polymer and a conductive material, wherein the second conductive polymer contacts the conductive material of the second sensor, wherein the resistivity of the second conductive polymer is responsive to 12 CO2 and 13 the concentration of CO2 decreases as it increases, wherein the second conductive polymer comprises sulfonated polyaniline blended with polyethylene oxide; processor; and circuit, wherein the circuit operatively connects the first and second sensors to the processor, wherein the processor detects resistivity in the circuit and uses the resistivity to calculate the total concentration of ammonia in the breath sample and 12 CO2 and 13 Total CO2 concentration.

2. The breath analyzer according to claim 1, wherein The sulfonated polyaniline is synthesized from an emeraldine form of polyaniline polymer.

3. The breath analyzer according to claim 1 or claim 2, wherein: The conductive material of the first sensor and / or the second sensor comprises a plurality of electrodes.

4. The breath analyzer according to claim 3, wherein The plurality of electrodes includes conductive wires arranged in a helical configuration.

5. The breath analyzer according to claim 3, wherein The plurality of electrodes include conductive lines arranged in a rectangular configuration.

6. The breath analyzer according to any one of claims 1 to 5, wherein: The breath analyzer is in wireless communication with a remote device.

7. The breath analyzer according to any one of claims 1 to 5, wherein: The breath analyzer has two separate channels through which the breath sample is configured to pass from the input to each of the first and second sensors.

8. The breath analyzer according to any one of claims 1 to 5, wherein: The ratio of polyethylene oxide to sulfonated polyaniline is 10 to 30 weight %.

9. The breath analyzer according to claim 8, wherein The ratio of polyethylene oxide to sulfonated polyaniline was about 30 wt%.

10. A method for detecting Helicobacter pylori in a subject's digestive tract, the method comprising: collecting a baseline breath sample from a subject using the breath analyzer of any one of claims 1 to 9; Determine the total amount of ammonia present in the baseline breath sample using the breath analyzer and 12 CO2 and 13 Total amount of CO2; The subjects were allowed to take a 13 C-labeled or unlabeled urea diet; collecting a post-urea ingestion breath sample from the subject using the breath analyzer; The breath analyzer was used to determine the total amount of ammonia present in the breath sample after urea ingestion and 12 CO2 and 13 Total amount of CO2; as well as If the total amount of ammonia in the breath sample after urea ingestion is 12 CO2 and 13 The total amount of CO2 exceeds the total amount of ammonia present in the baseline breath sample by a predetermined value and 12 CO2 and 13 The total amount of CO2 indicates the presence of Helicobacter pylori in the subject's digestive tract.

11. The method according to claim 10, further comprising: If the total amount of ammonia present in the breath sample after the urea ingestion and 12 CO2 and 13 The total amount of CO2 does not exceed the total amount of ammonia present in the baseline breath sample by the predetermined value and 12 CO2 and 13 The total amount of CO2 indicates that Helicobacter pylori is not present in the subject's digestive tract.

12. The method of claim 10 or claim 11, wherein collecting the baseline breath sample from the subject and collecting the post-urea ingestion breath sample from the subject comprises collecting the baseline breath sample and the post-urea ingestion breath sample from a single subject and from a single portable breath analyzer, the single portable breath analyzer being the breath analyzer of any one of claims 1-9.

13. A breath test method comprising the following steps: (a) providing a breath analyzer as claimed in any one of claims 1 to 9; (b) prompting the subject to exhale a baseline breath sample into the breath analyzer; (c) allowing the processor to measure the resistivity of the first sensor as it occurs when the baseline breath sample contacts the first sensor; (d) allowing the processor to measure the resistivity of the second sensor that occurs when the baseline breath sample contacts the second sensor; (e) providing the subject with a meal or capsule containing urea, wherein the urea is 13 C-labeled or unlabeled; (f) prompting the subject to exhale the post-urea breath sample into the breath analyzer; (g) allowing the processor to measure the resistivity of the first sensor as it occurs when the post-urea breath sample contacts the first sensor; (h) allowing the processor to measure the resistivity of the second sensor as it occurs when the post-urea breath sample contacts the second sensor; (i) comparing the measured resistivity of the baseline breath sample with the measured resistivity of the post-urea ingestion breath sample; (j) calculating a difference between the resistivity of the first sensor of the baseline breath sample and the measured resistivity of the first sensor of the post-urea ingestion breath sample; (k) calculating a difference between the resistivity of the second sensor of the baseline breath sample and the measured resistivity of the second sensor of the post-urea ingestion breath sample; (1) Express the resistivity difference of the first sensor in ppb of NH3 and the resistivity difference in CO2 or 13 The resistivity difference of the second sensor is expressed in ppm of CO2; and (m) When the difference between the resistivity measured after urea and the baseline of the first sensor and the second sensor is a positive number, the final result is displayed as Helicobacter pylori positive; when the difference between the resistivity measured after urea and the baseline of the first sensor and the second sensor is zero or a negative number, the final result is displayed as Helicobacter pylori negative.

14. The breath test method according to claim 13, wherein: The urea is unlabeled urea.

15. The breath test method according to claim 13 or claim 14, wherein: The subject exhaled the urea post-ingestion breath sample into the breath analyzer within 10-90 minutes after ingesting the meal or capsule.

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