WO2021245565A1 - Nucleodx rapid test - Google Patents

Abstract

The invention is for a point-of-care diagnostic device for detection of one or more biological targets of interest in a sample obtained from a patient and diagnosis of the patient, the device comprising: a reaction tube receiving chamber for receiving a single reaction tube that is associated with a radiofrequency identification (RFID) tag, the sample being received in the reaction tube and a biological reaction being conducted therein, when the reaction tube is received within the chamber; a sample sterilizer in communication with the reaction tube chamber configured to sterilize the sample by killing or deactivating microbes in the sample; a gesture sensor configured to receive a contactless user input from a user to initiate the biological reaction and obtain a diagnostic result of the sample; a thermal sensor for measuring and monitoring temperature of the sample during the biological reaction; an RFID reader configured to read the RFID tag associated with the reaction tube; a display screen configured to display the diagnostic result; and a wireless communication device configured to send the diagnostic result and patient data to another device. The invention also discloses a method of using the diagnostic device.

Description

NUCLEODX RAPID TEST

FIELD OF THE INVENTION

THIS INVENTION relates to a real-time point-of-care diagnostic device for the identification of one or more biological targets of interest of a biological or synthetic organism which inter alia, allow for diagnosis of a disease, disorder or condition, or the identification of specific attributes in a sample.

BACKGROUND TO THE INVENTION

Given the current external environment vis-a-vis the worldwide pandemic, the specific application of the device to diagnose for COVID-19, caused by the SARS-CoV-2 virus, is discussed as an example. It must be noted that the device, however, can be used to diagnose a range of diseases, disorders and conditions by testing for the presence of various biological targets.

In this specification, the term “disease” means a condition of the living animal or plant body or of one of its parts that impairs normal functioning and is typically manifested by distinguishing signs and symptoms.

In this specification, the term “disorder” means an illness that disrupts normal physical or mental functions.

In this specification, the term “condition” means a subject’s state of health which is an abnormal state that feels different from a subject’s normal state of wellbeing.

In this specification, the term “biological targets” means living or dead biological material that may have harmful effects for humans or animals, and includes, amongst others, genetic material, microorganisms (such as bacteria, fungi, viruses and parasites), toxins, allergens, hormones, proteins.

The COVID-19 pandemic has highlighted the need for diagnostic devices and tests. Most of the approved diagnostic workflows require RNA extraction and/or real-time reverse transcription PCR (RT-PCRs) for viral diagnosis which has resulted in global shortages. This has led to a large number of new tests being developed for COVID- 19 using alternative, previously proven, diagnostic technologies.

Isothermal amplification technologies (iNAT or iNAAT) are good alternatives to PCR- based amplifications assays and only require simple instrumentation. The technology has previously been used in diagnostic reverse transcription loop-mediated isothermal amplification (RT-LAMP) tests for MERS-CoV. A number of RT-LAMP diagnostic protocols have been developed for COVID-19 testing and show that RT-LAMP assays are comparable to the standard RT-PCR tests, often faster than these, and could be run directly from a sample without prior RNA extraction, while targeting multiple viral regions. It has been shown that RT-LAMP reactions can be multiplexed and visualized using sequence-specific fluorescent probes (oligonucleotide strand exchange probes). In this manner, multi-regional redundant assays and sequence specific readouts reduced false negatives. This corresponds to the current RT-PCR diagnostic tests and recommendations to target multiple genes (usually 2) within the SARS-CoV- 2 genome. For this, targets included the nucleocapsid, spike, RNA-dependent RNA polymerase (RdRP)/helicase, RdRP-P2, ORF1ab, and other genes.

The RT-LAMP technology offers a faster and low-resource alternative to RT-PCR based COVID-19 diagnostic tests. It allows for the reaction to be conducted in a single tube without the need for RNA extraction and targets multiple conserved regions in the genome of interest. By targeting two or three regions of the genome, a (positive) test redundancy is provided to accommodate for any mutation in a priming site that might result in a false negative.

Previously, RT-LAMP diagnostics lacked a reaction control needed to verify a true negative result. Reaction controls enable reaction verification during the test, i.e., a positive reaction control confirms that workflow steps (the RNA extraction and/or RT- PCR reactions) were successful even if the viral pathogen test result was negative. This prevents failed reactions or degraded samples being interpreted incorrectly as a negative result. The COVID-19 diagnostic tests targeting different genomic regions are run concurrently with the reaction control in the same reaction tube for each sample being assessed. Historically, RT-LAMP diagnostic applications targeted a single region and provided a positive/negative answer that is endpoint visualised. However, new sequence specific RT-LAMP target identification approaches now enables multiplexing and includes a reaction control.

RT-LAMP reactions are often used in a yes/no testing manner, using easily detectable visualisation dyes (and other methods) linked to nucleic acid accumulation. The use of multiple dyes in RT-LAMP, furthermore, allows for the identification of different testing regions. It is therefore possible to use different dyes, each linked to a viral gene target or internal human control gene product, to not only increase the sensitivity and specificity of the test, but also to lower false negatives. RT-LAMP technology is known for its low-cost, low-infrastructure endpoint result detection, with scanners, cell phones or even visual confirmation by eye, often used after an isothermal heating step (Table 1 ). Additionally, there are a number of studies presenting the use of fluorescent dyes in DIY applications on microscopy and other fields.

Although there have been a number of different testing methodologies released and accredited since the COVID-19 virus’ occurrence at the end of 2019, many of these workflows are either aimed at laboratory implementation or require specialized equipment and/or chemistries that limit wide point-of-care (POC) implementation. Current limitations in diagnostics include low viral load in samples, collection difficulties of samples from patients, inappropriate sample handling, delays causing RNA sample degradation (e.g., transport, storage) and insufficient sample loading during RT-qPCR tests. It has been shown that the SARS-CoV-2 virus can be detected consistently in-patient saliva (3.3 x 10⁶ copies/ml), and it has further been shown that even lower loads were sufficient for COVID-19 LAMP assays. Together with the device claimed here, this can greatly facilitate onsite or at home testing.

By taking advantage of the isothermal nature of RT-LAMP reactions that allows for easy implementation of the technology in resource poor environments, as well as its robustness to work straight from a sample without the need of a RNA extraction, the RT-LAMP assay can be conducted on the invention’s simple point-of-care device which provides the required heat and detection.

DISCLOSURE OF THE INVENTION The invention discloses the development of a portable, point-of-care (POC) device that utilises enzymatic reactions, such as the RT-LAMP assay, strand displacement reactions and digestions and integrates this with radio-frequency identification (RFID) technology, automatic testing and inactivation of samples to conduct and present the results of a detection test, such as a nucleotide (DNA or RNA) specific test. The device obviates the need for a laboratory and/or sterile environment and results are provided in real-time and are securely managed. The device therefore produces accurate, safe and secure diagnostic results.

The device is an electronic device that measures a range of biological targets, such as the nucleotides DNA and RNA, using wide band light frequency spectrometric measurement and narrow band light frequency spectrometric measurement. The device can be used to measure, for example, specific viral presence, specific bacterial presence, specific fungi presence, presence of specific chemicals, presence of specific minerals, presence of specific vitamins, specific blood presence, presence of specific fluids, and for various properties and possibilities of each. It can be used to detect at least one biological target or multiple, for example it can detect a single genetic region of interest or can detect multiple genetic regions of interest in a single sample.

The device can perform testing for diagnostic purposes, for example nucleotide testing, such as is required in the medical, zoological, forensic, water quality, food safety, and other fields. It is a user-operatable device to be used by adult literate individuals, but can also be used in a formal clinical environment i.e., doctors’ rooms, pharmacies, laboratories, mobile test facilities, or other fields where an onsite diagnostic may be required, e.g., onsite food safety testing, infield water quality testing, etc.

The device provides simplistic, live, aminated demonstrations on a wide screen with audio assistance as well as an operator manual that allows for easy operation and guidance during test procedures. Its robust and inclusive design allows for it to be operated effectively with a minimum of training and does not require a high level of technical expertise. It has testing, diagnostic and research functionality. The device can be used in a number of fields for testing, assays, or diagnostics such as single to multi-virus detection, pathogen screening, disease confirmation and mutation detection where it is used to test for specific genetic fragments or combinations thereof.

The patient is rapidly enrolled on the device prior to approval of the test. The patient must accept or reject the terms and conditions using the motion detection function on the screen. The database of the device is linked to a mobile app or a web-based (online) link to allow for the loading of personal data, consent to the diagnostic, agreement to terms, selection of tests, payment, etc.to take place. This link from a phone or smart ID to the device is active at the point of testing and further removes the need for direct interaction with the device.

The device provides real time reporting to the required authority or regulatory organization (such as the National Health Laboratory Service, Covid Watch and World Health Organization). Results are released to the relevant body depending on the appropriate authority or offsite server confirmation, thereby allowing for automatic reporting when and/or where mandatory. The information conveyed to the relevant body may include, amongst others, the biochemistry for a specific test, origin and expiry date of the biochemicals in the test tube, location of the test, personal information of the patient being tested, medical symptoms of the patient, a digital picture of the patient, biometrics from the picture, test results and consent of the patient. Patient information is encrypted and/or tracked and/or compiled (e.g., blockchain) to ensure confidentiality and protection of the patient’s rights and legal compliance with prevailing local and international privacy and information disclosure legislation.

The device includes a unique and accurate sampling system that ensures consistent sample sizes for testing. This system includes sample tracking. The system can also use current sampling equipment. The risk of contamination is minimized to the user. The screen includes motion detection, thus removing the need to touch the screen. All samples are automatically treated with a built-in ultraviolet-C (UVC) treatment system in the waste bin. The device is fast to develop or convert, easy to implement, has a history of previous diagnostic application, but is sensitive enough to be informative in the real-time detection and diagnosis of RNA or DNA in samples.

The sample may be in the form of any suitable bodily fluid, such as saliva. The bodily fluid such as saliva is placed in an automatable sample tube and circumvents the swab-to-96-well bottleneck in the RT-PCR workflow. The device runs the test directing on the sample which further expedites the workflow. When a test is run where the reaction is isothermal, the device provides the required heating. The reactions also require low-end processing and thus the POC device is ideal and provides on-site results.

The invention is able to run different tests, such as the RT-LAMP assay, to expedite diagnostic tests and utilises resources not used in current standard RT-PCRs, thus lowering the technology needed for implementation, yet retaining a comparable diagnostic sensitivity and specificity. The point-of-care or at-home test allows for rapid detection of genetic material of disease-associated biological material, such as the SARS-CoV-2 virus, in a bodily fluid sample and provides dye-based visualisation of such genetic material. The device can be used by individuals at home for point-of-care testing, enabling said individuals to test themselves using easily obtainable saliva or a similar sample type as the starting point. By allowing fast, accurate and at home or POC site testing, larger numbers of individuals can screen themselves more frequently and most likely cheaper. The test would be faster and easier to implement on sites (i.e., at businesses, homes, schools, etc.). This approach aims to remove the need to travel during testing activities, thereby removing the exposure of health care professionals and the virus spreading. The use of saliva, for example, makes the test easier to conduct by the general public and does not require an RNA extraction prior to testing, removing the need for onsite RNA extraction infrastructure and the transport of possible infectious materials to central testing facilities. Additionally, direct testing removes possible biological degradation in samples prior to testing. Since the reaction sample tube is only open to add the bodily fluid sample for testing, possible amplicons and the spread of viruses and bacteria is minimised. The device analyses the sample on the basis of biological, chemical, electrical, thermal, optical, and structural properties, using fluid, acids, light, colour, gas, temperature, energy radiation, microbes, compositions, activators, stimulators, and excitation elements. Secondary input factors that are taken into consideration include body temperature, blood oxygenation, pain, discomfort, colour, infection, symptoms, blood pressure, reaction, and structure. Such assessment utilizes genetic target fragments, biochemicals, spectrometric and/or gas chromatic excretion principles to form an output measurement pattern, and a result that is electronically measured and structured into a simple outcome or as a comprehensive detailed result. One or several sampler sections can be loaded in no particular order for each sample section’s own assessment process.

The device can be used to measure or determine the presence of select genetic regions in the genome of an organism of interest. It can also assess tissue, structure, and other properties of the sample, it measures biochemical and electrical properties and behaviour patterns in the sample. Such information gathered can be used for research and development and commercial applications. The device provides the options to measure the presence and properties of particular elements: measure mutations and their properties; stimulate specific activities; measure levels, properties, behaviour, and potential of components of a composition; and observe biochemical factors and properties.

The device uses the following principles in assessment or analysis of the sample: measurement of colour and/or frequency spectral properties; measurement of gas presence and/or formation; measurement and/or observation of structural biological, chemical and biochemical formation; measurement of energy activation, behaviour, excretion, radiation and or absorption properties; measurement of chemical reaction properties; measurement of biological behaviour and/or reaction properties; consideration of external influential factors, in this specification, the term “spectrometry” means the measurement of the interactions between light and matter and includes colorimetric and fiuorometric detection. The device utilizes unique spectrometric measurement by using three narrow-band light sources in addition with a Red Green Blue (RGB) light source combined with a white light source to cover the entire spectrum of illumination for a wide spectrum of applications and research options. White light provides a universal light source and/or for the detection of multiple colours in spectrometry. The specific light sources are selected for detection of specific colours in spectrometry. The device includes the following features:

1. RFID reading;

2. Camera for biometrics and/or telemetries;

3. Thermal sensor for telemetries;

4. Gesture control for touchless control; 5, Sample sterilization;

6. Wireless communication means, such as Wi-Fi USB and/or Bluetooth communication;

7. PC and/or remote device control software. The device forms part of a system that consists of: a container; a motorized sample door; a sample tube receiving and ejection mechanism; a sample tube guiding structure; a tested sample ejection guide; a tested sample drawer or containment bin; a drawer sensing switch; a UVC sterilizing light source; a UVC sensor; power supply; a LCD touch screen; a camera; an infra-red body temperature sensor; a wireless communication means {such as a Wi-Fi, RS-485 connection); USB port; on-off switch; micro-controllers; a sample heater; a sample heater temperature sensor; a sample cooling fan or cooling device; a RF tag reader; a gesture processor; an ambient light sensor; an RGB spectrum sensor; a visible light spectrometric sensing device; a narrow band red light source; a narrow band green light source; a narrow band blue light source; a white light source; a visible spectrum RGB light source; a SD card memory holder; internal memory; signal conditioner components; light brightness control components; stepper motor control circuit; and an indicator LED.

The device runs from a single 5V power source. The device can be powered from a battery as well as from a power source. The device is therefore portable.

USB communication connects the device to a computer. The computer can control the device. The computer can show the device’s functional processes. The computer can receive test results, store, or communicate the results and client data to the internet and to other external systems. A wireless communication means, such as Wi-Fi communication, is used to communicate results and client data to another device, such as the internet, to a remote server, such as in an airport, testing facility, hospital etc., or to each of various numbers of test modules that makes up a test system. RS-485 communication is used to communicate to each of various numbers of test modules that make supa test system.

The camera provides a photo of the patient being tested. The infrared heat sensor measures the patient’s body temperature or the object’s temperature.

UVC is used for killing or deactivating microbes that may be present in the test tubes after testing. A drawer switch indicates to the system when the UVC light may be activated. A UVC sensor indicates to the system that the UVC source is active.

A motorized structure opens the sample door for inserting the sample tube. The system will eject the sample tube after completing the test process or when the test tube fails to meet the process requirements. The ejected tube is guided to fall into the tested sample drawer or containment bin.

The gesture sensor provides a hands-free operation and instruction feature as a human interface. The RF tag reader reads a RF tag associated with the sample reaction tube or process for a unique serial number in order to track the sample reaction tube with the reagents used, the patient’s data and diagnostic results.

The ambient light sensor is used as part of the reference and calibration source.

Five types of sample excitation / illumination features can be applied, namely, narrow bands Red, Green, Blue; Full White light, and RGB mixed light. Narrowband light sources and wideband light sources are excitation sources for the wide spectrum of biochemical processes, each of which can be individually controlled. The RGB sensor and spectrometric sensors are utilized to measure the biochemical photonic properties of the test sample. The device includes a sample heater and cooler where the temperature controller and measurement system form part of the test process.

The LCD display touch screen provides promotional and motivational information, objective information, legal information, process information, user instructions, process details, acts as a user control interface, displays test results, provides user options information, provides relevant feedback data, and provides visual support information and education data.

Further applications of the device include, for example, the research of plant extracts, soil minerals and microfiora; the identification of specific biological factors associated with maternal health risk and intervention in livestock; the identification of health- associated factors in livestock; measurement of factors associated with disease treatment in a diseased animal to track progress and recovery; measurement of biological factors to test the sweetness of a fruit.

According to a first aspect of the invention there is provided a point-of-care diagnostic device for detection of one or more biological targets of interest in a sample obtained from a patient and diagnosis of the patient based on a diagnostic result of the sample, the device comprising the following: a reaction tube receiving chamber for receiving a single reaction tube that is associated with a radiofrequency identification (RFID) tag, wherein the sample is received in the reaction tube and a biological reaction is conducted therein, when the reaction tube is received within the chamber; a sample sterilizer in communication with the reaction tube chamber configured to sterilize the sample by killing or deactivating microbes in the sample; a thermal sensor for measuring and monitoring temperature of the sample during the biological reaction; an RFID reader configured to read the RFID tag associated with the reaction tube for sample tracking; a display screen configured to display the diagnostic result; and a wireless communication device configured to send the diagnostic result and patient data to another device.

In this specification, the term “sample tracking” includes confirming the type of reaction to be conducted and biochemical logistical history.

The one or more biological targets may include genetic mutations in a genome of an organism, or biological structures in an organism.

The one or more biological targets may include one or more genetic fragments of a genome of an organism.

The device may identify the presence of one or more genetic fragments in the absence of RNA extraction.

The one or more biological targets may be detected by DNA or RNA testing using a nucleic-acid based assay, including an RT-LAMP assay.

The sample may be bodily fluid, including at least one of saliva, breath, tissue and blood, obtained from the patient.

The device may identify the presence of a virus in the sample, including a SARS-CoV- 2 virus for diagnosis of COVID-19.

The diagnostic result may be detected using spectrometry, including colorimetric or fluorometric detection.

The device may include at least one narrowband excitation source that provides light for the detection using spectrometry.

The device may include three narrowband excitation sources, a Red Green Blue (RGB) light source and a white light source that provide light for the detection using spectrometry. The device may include an RGB sensor and a spectrometric sensor to measure biochemical photonic properties of the sample. The device may include an ambient light sensor configured to measure ambient light, such ambient light measurement may be configured to be used for calibration of the RGB sensor and the spectrometric sensor.

The sample sterilizer may be an ultraviolet-C sterilizer and may include a sensor configured to detect microbe levels in the sample.

The display screen may be a liquid crystal display (LCD) touch screen and may be configured both to display the diagnostic result and to receive a touch-based user input.

The device may include an infrared body temperature sensor to sense a body temperature of the patient and the sensed body temperature may form part of the diagnostic result. The device may include a sample heater and cooler to control the temperature of the sample.

The device may include a sample tube guiding structure and a sample tube ejecting mechanism for ejecting tested sample tubes into a containment bin.

The device may include a camera for capturing a photograph of the patient.

The device may include an RFID writer configured to write the diagnostic result of the sample to the RFID tag that is associated with the sample reaction tube.

The RFID tag that is associated with the reaction tube may be a first RFID tag and the RFID reader may be configured to read a second RFID tag associated with a swab used to collect the biological sample, wherein information read from the second RFID tag forms part of the diagnostic result. The device may include a gesture sensor configured to receive a contactless user input from a user to initiate the biological reaction and obtain the diagnostic result of the sample.

According to a second aspect of the invention, there is provided a method of operating a point-of-care diagnostic device for real-time detection of one or more biological targets of interest in a sample obtained from a patient and diagnosis of the patient based on a diagnostic result of the sample, the method comprising: receiving a single reaction tube in a reaction tube receiving chamber, wherein the single reaction tube is associated with a radiofrequency identification (RFID) tag, the sample being received in the reaction tube and a biological reaction being conducted therein, when the reaction tube is received within the chamber; sterilizing, by a sample sterilizer in communication with the reaction tube chamber, the sample by killing or deactivating microbes in the sample; measuring, by a thermal sensor, temperature of the sample during the biological reaction; reading, by an RFID reader, the RFID tag associated with the reaction tube for sample tracking; displaying, by a display screen, the diagnostic result; and sending, by a wireless communication device, diagnostic result and patient data to another device.

The invention will now be described in more detail, with reference to the following non- limiting example and the accompanying drawings.

In the drawings

FIGURE 1 shows different views of the frame of the device; and FIGURE 2 shows the frame of the device indicating where the components are received.

With reference to Figure 2, the device (10) includes an LCD touch screen (12), a sample loader mechanism (14), a temperature sensor (16), a camera (18) and a samples drawer (20). EXAMPLE: Use of the device to test for COVID-19 genetic regions using an RT-

LAMP assay

Multiplexed LAMP assays that target multiple SARS-CoV-2 genetic regions and an internal reaction control gene RT-LAMP test are utilised, together with sequence- specific fluorescently labelled probes, as a faster and cheaper diagnostic assay than the standard RT-qPCR workflow. This has been shown to be possible for COVID-19 diagnostics by a number of studies, even with multiple viral regions being targeted (Table 1 ). Multiple viral targets address any RNA virus genetic variabilities that may result in mismatching of assay primers and probes, thus providing a false negative, if only a single gene is targeted in an assay. Standard colorimetric assays that are easy to interpret by colour, will be the first assay for a fast yes/no assessment (i.e., not taking reagent or sample viability into account). However, RT-qPCR diagnostic test often utilizes a constitutively expressed human gene as a workflow or reaction validation control. A suitable RT-LAMP assay is developed that targets one of these “in-sample” human gene regions and/or an external RNA standard added to the reaction mix, as an in-reaction viability confirmation. The SARS-CoV-2 gene targets and the in-reaction viability assay are labelled with different labels to facilitate individual reaction confirmation.

A positive DNA control LAMP reaction is used as a standard and will convert an RNA template into a more robust DNA template in one of the assays, to conduct the device optimisation. Colorimetric assessment is included for yes/no assays.

Samples of bodily fluids are utilised, such as saliva, breath, body odour, tissue, blood, etc. Such sample will be collected by means of paper, stick, spoon, pipette, syringe, disc, needle, flask, bottle, tube, or similar. A saliva sample in-a-tube is easy to obtain, removing the need for swabs and enabling easier self-testing applications and/or liquid handling automation.

The RT-LAMP assay includes the use of SARS-CoV-2 RT-LAMP primers and probes. Positive control DNA/RNA fragments will be synthesized for testing the different gene targeting RT-LAMP primers and will include features to allow for in vitro RNA template generation or will include synthesized RNA fragments for workflow testing and quality control purposes. Non-BSL extracted RNA of SARS-CoV-2 would serve as spikes for human samples (e.g., saliva).

A standard human housekeeping gene is used as a reaction verification RT-LAMP control. Fluorescent probes and quenchers allow for target specific RT-LAMP product identification. The fluorescent probes are robust enough to distinguish between a COVID-19 test and the internal reaction control. This will initially be validated on a real time PCR system to ensure a working chemistry prior to use in the POC device.

Discussion

The invention is directed at a diagnostic point-of-care device for the identification of biological targets of interest from a biological or synthetic organism of interest for onsite, or at home, diagnosis of diseases, disorders and conditions in real-time. The device reduces assay times and removes the need for central processing. The device may identify, for example, the presence of one or more genetic regions of interest of a genome of a microorganism, such as a virus or bacteria, or the presence of one or more genetic regions of interest or genetic mutations of interest that are associated with a particular disease, disorder or condition. The device alleviates the problems associated with diagnosis by providing a portable device that runs off a simple power source, such as a battery pack; requires a bodily fluid sample that can be easily obtained from a patient; uses a single reaction tube for collecting and running a reaction from a sample; runs the biological reaction entirely within the device and does not require intervention from a user; maintains patient confidentiality; tracks the sample using RFID technology; obtains and presents the diagnostic results in realtime; and conveys such diagnostic results to a user or relevant authority. Bhadra S, Riedel TE, et al. (2020). High-surety isothermal amplification and detection of SARSCoV-2, including with crude enzymes. bioRxiv2020.04.13.039941; doi: https://doi.org/10.1101/2020.04.13.039941 El-Tholoth M, Bau HH, et al. (2020). A Single and Two-Stage, Closed-Tube, Molecular Test for the 2019 Novel Coronavirus (COVID-19) at Home, Clinic, and Points of Entry. ChemRxiv. Preprint. https://doi.org/10.26434/chemrxiv.11860137.v1 Lamb LE, Bartolone SN, et al. (2020). Rapid Detection of Novel Coronavirus (COVID19) by Reverse Transcription-Loop-Mediated Isothermal Amplification. https://ssrn.com/abstract=3539654 or http://dx.doi.org/10.2139/ssrn.3539654 Lee JYH, Best N, et al. (2020). Validation of a single-step, single-tube reverse transcription-loop mediate isothermal amplification assay for rapid detection of SARS- CoV-2 RNA. bioRxiv2020.04.28.067363; doi: https://doi.org/10.1101/2020.04.28.067363 Lu R, Wu X, Wan Z, et al. (2020). Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. Virol Sin.1‐4. doi:10.1007/s12250-020-00218-1 Lu R, Wu X, et al. (2020). A Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2. Int J Mol Sci. 2020; 21(8):E2826. Published 2020 Apr 18. doi:10.3390/ijms21082826 Osterdahl MF, Lee KA, et al. (2020). Detecting SARS-CoV-2 at point of care: Preliminary data comparing Loop-mediated isothermal amplification (LAMP) to PCR. medRxiv2020.04.01.20047357; doi: https://doi.org/10.1101/2020.04.01.20047357 Park GS, Ku K, et al.(2020). Development of Reverse Transcription Loop-Mediated Isothermal Amplification Assays Targeting Severe Acute Respiratory Syndrome https://doi.org/10.1016/j.jmoldx.2020.03.006 Yan C, Cui J, et al. (2020). Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay. Clinical microbiology and infection 26 (6): 773-779. doi:10.1016/j.cmi.2020.04.001 Yang W, Dang W, et al. (2020). Rapid Detection of SARS-CoV-2 Using Reverse transcription RT-LAMP method. medRxiv2020.03.02.20030130; https://doi.org/10.1101/2020.03.02.20030130 Yu L, Wu S, et al. (2020). Rapid colorimetric detection of COVID-19 coronavirus using a reverse transcriptional loop-mediated isothermal amplification (RT-LAMP) diagnostic platform: iLACO. medRxiv2020.02.20.20025874; https://doi.org/10.1101/2020.02.20.20025874 Zhang Y, Odiwuor N, et al. (2020). Rapid Molecular Detection of SARS-CoV-2 (COVID-19) Virus RNA Using Colorimetric LAMP. medRxiv2020.02.26.20028373;doi: https://doi.org/10.1101/2020.02.26.20028373

Claims

CLAIMS:

1 . A point-of-care diagnostic device for detection of one or more biological targets of interest in a sample obtained from a patient and diagnosis of the patient based on a diagnostic result of the sample, the device comprising: a reaction tube receiving chamber for receiving a single reaction tube that is associated with a radiofrequency identification (RFID) tag, the sample being received in the reaction tube and a biological reaction being conducted therein, when the reaction tube is received within the chamber; a sample sterilizer in communication with the reaction tube chamber configured to sterilize the sample by killing or deactivating microbes in the sample; a thermal sensor for measuring and monitoring temperature of the sample during the biological reaction; an RFID reader configured to read the RFID tag associated with the reaction tube for sample tracking; a display screen configured to display the diagnostic result; and a wireless communication device configured to send the diagnostic result and patient data to another device.

2. The device of claim 1 , wherein the one or more biological targets includes genetic mutations in a genome of an organism, or biological structures in an organism.

3. The device of claim 1 , wherein the one or more biological targets includes one or more genetic fragments of a genome of an organism.

4. The device of claim 3, wherein the device identifies the presence of one or more genetic fragments in the absence of RNA extraction.

5. The device of any one of claims 1 to 4, wherein the one or more biological targets are detected by DNA or RNA testing using a nucleic-acid based assay, including an RT-LAMP assay.

6. The device of any one of claims 1 to 5, wherein the sample is bodily fluid, including at least one of saliva, breath, tissue and blood, obtained from the patient.

7. The device of any one of claims 1 to 6, wherein the device identifies the presence of a virus in the sample, including a SARS-CoV-2 virus for diagnosis of COVID-19.

8. The device of any one of claims 1 to 7, wherein the diagnostic result is detected using spectrometry.

9. The device of claim 8, wherein the device includes at least one narrowband excitation source to provide light for the detection using spectrometry.

10. The device of claim 9, wherein the device includes three narrowband excitation sources, a Red Green Blue (RGB) light source, and a white light source to provide light for the detection using spectrometry.

11. The device of any one of claims 8 to 10, wherein the device includes an RGB sensor and a spectrometric sensor to measure biochemical photonic properties of the sample.

12. The device of claim 11 , wherein the device includes an ambient light sensor configured to measure ambient light, such ambient light measurement configured to be used for calibration of the RGB sensor and the spectrometric sensor.

13. The device of any one of claims 1 to 12, wherein the sample sterilizer is an ultraviolet-C sterilizer and includes a sensor configured to detect microbe levels in the sample.

14. The device of any one of claims 1 to 13, wherein the display screen is a liquid crystal display (LCD) touch screen, thus being configured both to display the diagnostic result and to receive a touch-based user input.

15. The device of any one of claims 1 to 14, which includes an infrared body temperature sensor to sense a body temperature of the patient, the sensed body temperature forming part of the diagnostic result.

16. The device of any one of claims 1 to 15, wherein the device includes a sample heater and cooler to control the temperature of the sample.

17. The device of any one of claims 1 to 16, wherein the device includes a sample tube guiding structure and a sample tube ejecting mechanism for ejecting tested sample tubes into a containment bin.

18. The device of any one of claims 1 to 17, wherein the device includes a camera for capturing a photograph of the patient.

19. The device of any one of claims 1 to 18, wherein the device includes an RFID writer configured to write the diagnostic result of the sample to the RFID tag that is associated with the sample reaction tube.

20. The device of any one of claims 1 to 19, wherein the RFID tag associated with the reaction tube is a first RFID tag and the RFID reader is configured to read a second

RFID tag associated with a swab used to collect the biological sample, wherein information read from the second RFID tag forms part of the diagnostic result.

21. The device of any one of claims 1 to 20, wherein the device includes a gesture sensor configured to receive a contactless user input from a user to initiate the biological reaction and obtain the diagnostic result of the sample.

22. A method of operating a point-of-care diagnostic device for real-time detection of one or more biological targets of interest in a sample obtained from a patient and diagnosis of the patient based on a diagnostic result of the sample, the method comprising: receiving a single reaction tube in a reaction tube receiving chamber, the single reaction tube being associated with a radiofrequency identification (RFID) tag, the sample being received in the reaction tube and a biological reaction being conducted therein, when the reaction tube is received within the chamber; sterilizing, by a sample sterilizer in communication with the reaction tube chamber, the sample by killing or deactivating microbes in the sample; measuring, by a thermal sensor, temperature of the sample during the biological reaction; reading, by an RFID reader, the RFID tag associated with the reaction tube for sample tracking; displaying, by a display screen, the diagnostic result; and sending, by a wireless communication device, diagnostic result and patient data to another device.