WO2019183739A1 - Mobile device that allows the detection of determined microorganisms in different types of samples - Google Patents

Mobile device that allows the detection of determined microorganisms in different types of samples Download PDF

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Publication number
WO2019183739A1
WO2019183739A1 PCT/CL2019/050022 CL2019050022W WO2019183739A1 WO 2019183739 A1 WO2019183739 A1 WO 2019183739A1 CL 2019050022 W CL2019050022 W CL 2019050022W WO 2019183739 A1 WO2019183739 A1 WO 2019183739A1
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WO
WIPO (PCT)
Prior art keywords
mobile device
allows
detection
samples
video
Prior art date
Application number
PCT/CL2019/050022
Other languages
Spanish (es)
French (fr)
Inventor
Francisco Javier ALTIMIRAS GONZÁLEZ
Leandro Anthony Emmanuel FARÍAS AGUILERA
Sebastián Alejandro GEERLING GAMBOA
Esteban Andrés GÁRATE ZUBIRI
Yassef Ariel YUIVAR VILLARREAL
Macarena Alejandra ARAYA TAPIA
Original Assignee
Protome Spa
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Publication date
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Publication of WO2019183739A1 publication Critical patent/WO2019183739A1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06F—ELECTRIC DIGITAL DATA PROCESSING
    • G06F18/00—Pattern recognition
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00—Image analysis
    • G06T7/10—Segmentation; Edge detection

Definitions

  • the present invention is part of the field of microbiological quality assurance, in particular in the detection of microorganisms in different types of samples.
  • the specific example of the field of application corresponds to the assurance of microbiological quality in the food industry, more specifically, in the monitoring and detection of microorganisms in an animal production plant.
  • the device of the present invention can be used in other industries that require monitoring and control of microbiological quality.
  • other areas where the device of the present invention could be used are: microbiological quality assurance of water, food processing plants, whether of vegetable or animal origin, among others.
  • WO2017025984A1 describes a device to be integrated into a smartphone, where the device allows isothermal nucleic acid amplification. It is specified that the phone adapts to the device, so that the camera of the phone can be used to record intensity measurements in micro-cameras (micro chambers) where the reaction is carried out.
  • CN 106010961 A describes a simple apparatus for the identification of pathogens in food, using as a base test loop-mediated isothermal amplification (Loop-mediated isothermal amplification, LAMP). It is mentioned that the device considers the use of a portable computer for the collection and processing of information. The apparatus considers a thermal plate, where the reaction is carried out, a fluorescent excitation device, a photography mechanism.
  • LAMP loop-mediated isothermal amplification
  • US2016231324A1 describes high performance throughput systems based on microfluidics.
  • one of the tests that can be adapted to the system is LAMP. He also mentions that the system can be designed to adapt it for portability, such as by including it in a backpack.
  • US2016216210A1 describes a portable device (hardware) for nucleic acid amplification, and software to be installed in a portable device to control and obtain the results generated by the hardware.
  • the nucleic acid amplification method can be LAMP.
  • WO2016064894A2 describes a method for detecting RNA viruses, which consists of a back-transcription reaction coupled with an isothermal amplification reaction. In the specification it is mentioned that the method can be carried out on a portable device at a point of care, however, it does not provide details of such a device.
  • US2015141264A1 describes a portable device for detection of specific DNA samples.
  • the claims indicate that one of the systems for DNA identification that can be implemented in the device is LAMP. It does not refer to pathogen identification, since the focus of this document is a system for authentication.
  • US2014335527A1 describes a mobile device for analysis of nucleic acids and proteins.
  • the device includes a module for nucleic acid extraction, and also for amplification (optionally it is indicated that it can be performed using, among other methods, LAMP).
  • the device is also equipped with wireless communication capabilities to transfer the results. No reference was found to a set of specific pathogens that could be detected with the system.
  • US2014220667A1 describes a portable device for pathogen detection.
  • LAMP is mentioned.
  • the device is quite different from what is proposed in the present invention, since it comprises a flexible strip comprising wells where aliquots of sample are deposited.
  • the text highlights the advantage of being a useful device for use in POC (point of care).
  • POC point of care
  • the application to specific pig pathogens is not mentioned.
  • Detection is performed the detection of different strains of E. coli.
  • CN 102199531 A describes a chip that uses microfluidics to detect several nucleic acid sequences based on a single sample. It is indicated that LAMP is used as the detection method. In the last figure they show an exemplary embodiment chip for detection of multiple pathogens in pigs. No reference was found to a mobile device for location use.
  • US2010273159A1 describes a method for the identification of multiple biological identities in a sample, comprising nested amplification reactions.
  • claim 9 the possibility of using an isothermal amplification method is indicated, and in 10 several alternatives are delivered, where LAMP appears.
  • W02006110855A2 describes methods for determining sequence variants.
  • the described method uses different rounds of amplification, where at least one of them is performed using an isothermal amplification.
  • LAMP is mentioned in the options for isothermal amplification.
  • the text of the document indicates that it can be applied to samples from different living beings, and it is mentioned, among many other options, to pigs.
  • There are differences with the present invention since it uses emulsion movable supports to immobilize some nucleic acids. In addition, it does not mention a mobile device to carry out the method.
  • EP3187595A1 describes a device for detecting Legionella strains in a sample.
  • the device includes heat sources, ultra violet (UV) light sources, and optical detectors.
  • UV ultra violet
  • US2016077006A1 describes a device that has heat sources, light sources, optical detectors, where the device can be used to detect nucleic acid amplifications.
  • US2014349298A1 describes an optoelectric device for pathogen detection using isothermal amplification. It is indicated that the device has a heat generating element, a UV light emitting diode, a sensor, the reading is communicated to an external device by means of wireless technology.
  • CN203720089U describes a device for sample analysis using isothermal amplification.
  • the device includes a heat source to control the temperature of the samples, a UV light source to read the result, and a UV light sensor (optical sensor).
  • the invention corresponds to a mobile device that allows to detect microorganisms in different samples, where the device works based on nucleic acid amplification reactions.
  • the present invention corresponds to a mobile device that allows the presence of multiple predetermined microorganisms to be detected in parallel, where the detection is carried out through the use of nucleic acid amplification techniques, combined with the use of probes specifically designed for the detection of said predetermined microorganisms.
  • the device comprises at least the following components: i. Controllable heat source, which ensures the maintenance of a predetermined temperature for a predetermined time, where the temperature control allows nucleic acid amplification reactions; ii. Support for sample container with multiple independent cameras, which allows depositing samples in each of the cameras;
  • Image capture device which allows obtaining an image of the support once the nucleic acid amplification reactions have been carried out
  • controllable heat source is selected from: a heating plate and a peltier plate.
  • the sample container holder corresponds to a perforated plate, where each perforation allows the positioning of each of the chambers present in the sample container.
  • the perforated plate material is selected from aluminum and copper.
  • the Perforated plate has a size between 80 mm and 130 mm per side, being able to have a square or rectangular shape.
  • the sample container with multiple independent chambers is selected from: 96-well plates, 8-tube strips, independent tubes. The volume of the individual tubes can vary between 20 and 500 pl.
  • the image capture device corresponds to a digital camera.
  • the camera allows to obtain an image of at least 5 megapixels of native resolution of the sensor.
  • the image capture device can also capture video with a resolution of at least 640x480 pixels at a frequency of between 60 and 90 frames per second.
  • video capture can be performed with a resolution of 720p at a frequency of 60 frames per second, and in yet another embodiment, video capture can be performed at 1080p at a frequency of 30 frames per second.
  • the lighting source is selected from a lighting source based on white light emitting diodes (Light-Emitting diodes, LEDs), UV, RGB, R, G or B, of type SMD, COB or high LED power, in which the wavelength of the light emitted by said LEDs corresponds to the entire visible range and UV, that is, in wavelengths between 395 to 700 nm.
  • the LED circuit operates with a voltage between 5 and 24 V, the total power dissipated by it between 4 and 20 W. The light intensity varies between 70 and 150 mcd per LED.
  • the invention also considers an LCD screen with a touch interface, where virtual or physical switches are presented, together with a knob-type regulator that allows, for example, to navigate through a menu of device options.
  • a resistive or capacitive TFT touch screen with a small HDMI interface between 2 and 9 inches is considered.
  • the resistive or capacitive TFT touch screen uses a SPI interface between 2 and 9 inches.
  • the acoustic and visual alarms correspond to audible or visible signals that are activated, for example, once the microorganism detection cycle has ended.
  • indicators are considered to indicate the power of the device with a preferably green light.
  • the indication of the heating process of the perforated plate is preferably performed with an intermittent yellow light between 5 and 30 min.
  • the signal at the end of the process is preferably performed with an intermittent green light and / or an intermittent acoustic signal (of a certain duration). This visual and / or auditory signal indicates to the user that the appropriate temperature has been reached to start the sample heating process, suggesting the user proceed to place the sample container on the perforated plate.
  • the device of the invention has a start button on the housing, which once pressed by the user, or triggered by a limit switch sensor, so that automatically and when placing the container of samples the device is activated and proceeds to start a timer so that the samples are heated for a certain amount of time, depending on the duration of the reaction. Once the new stage starts, the green LED would return to normal and cease to be intermittent.
  • a visual indicator preferably a red light and / or an acoustic signal, allows the user to suggest that the process is over and it is necessary to take the samples from the device.
  • a visual alert preferably a red light is activated intermittently.
  • the device also has a button or interface that allows to reset the process to start it again, or in the alternative, simply turn the equipment on and off.
  • the wireless communication media is selected from: Wi-Fi, 3G, LoRa.
  • the wireless communication means allows the information resulting from using the device of the present invention to be sent, that is, the capture of images or video.
  • the image or video information is sent to a remote server (virtual machine) in which the information obtained in the images or video is analyzed.
  • this analysis consists of the segmentation of the total image into singular units. Each singular unit corresponds to a replica of the sample taken and is compared, using a classifying algorithm, against at least two sets of manually curated images to determine which set each problem segment would belong to, which allows to know if the detection was positive or negative in the sample corresponding to that individual segment. Once all the individual segments have been classified, an automation is executed that stores the data and generates a report for the end user.
  • the data processed on the computer is displayed in one of the following ways: through a web application, to an application present on a mobile device, such as a smartphone, among others.
  • Example 1 Device connected to the cloud by Wi-Fi to measure nucleic acids.
  • the device allows diagnosing the presence of multiple pathogens in minutes (20-60 min).
  • the device consists of a 150 x 100 mm perforated copper and fiberglass heating plate that reaches temperatures of up to 100 ° C, a 12 V 8 Amp power source transformer, a 5 megapixel camera native resolution of the sensor with 2592 x 1944 pixels capacity, it is also compatible with 1080p30, 720p60 and 640x480p60 / 90 video, a lighting source designed from 12V UV LEDs, 12 W of power per meter and wavelength of 395 - 405nm, 2.4Ghz wireless capacity with a range of 60 meters indoors and up to 1200 meters outdoors.
  • the user deposits the sample in each of the detection wells.
  • the plate is heated and the isothermal amplification reaction is performed. After 20-60 min (at 60 ° C), the plate is illuminated with UV light and an image capture is performed. The image is sent via Wi-Fi to a remote computer for analysis and storage in a database.
  • the analysis is performed using an image segmentation algorithm and a classification algorithm based on colorimetry to determine whether the result of each individual reaction is positive or negative.
  • Example 2 Detection of Staphylococcus aureus from swine nasal samples
  • samples are taken from the porcine nostrils using a sterile torula, which is preserved, in an Eppendorf tube with a DNA stabilizing reagent (DNA Shield).
  • the samples are mixed with a rapid DNA extraction solution (20 min), using a stirring and heat protocol.
  • the samples are added in a 96-well plate, which already contains the corresponding reagents the kit to detect this pathogen by isothermal DNA amplification (enzyme, dNTPS, splitters, buffer and fluorescent or colorimetric marker).
  • the protocol for adding the samples to the plate consists of loading 10 pL from position D4 to G9, filling in total 36 wells of the 96-well plate (see Fig.
  • the present invention has application in any industry that requires monitoring or determining the presence of specific microorganisms in different types of samples.
  • a specific industry for example, corresponds to the animal production industry for human consumption.
  • a specific application is in the detection of pathogens that produce respiratory diseases in production pigs.

Abstract

The present invention relates to the field of microbiological quality assurance, particularly to the detection of microorganisms in different types of samples, and concerns a mobile device that allows the presence of multiple predetermined microorganisms to be detected in parallel, the detection being carried out using nucleic acid amplification techniques in combination with the use of probes specifically designed to detect said predetermined microorganisms.

Description

DISPOSITIVO MÓVIL QUE PERMITE LA DETECCIÓN DE MICROORGANISMOS DETERMINADOS EN DISTINTOS TIPOS DE MUESTRAS  MOBILE DEVICE THAT ALLOWS THE DETECTION OF MICROORGANISMS DETERMINED IN DIFFERENT TYPES OF SAMPLES
CAMPO TÉCNICO TECHNICAL FIELD
La presente invención se enmarca en el campo de aseguramiento de calidad microbiológica, en particular en la detección de microorganismos en distintos tipos de muestras. The present invention is part of the field of microbiological quality assurance, in particular in the detection of microorganisms in different types of samples.
ANTECEDENTES Y ARTE PREVIO BACKGROUND AND PRIOR ART
En la descripción de la presente invención, que corresponde a un dispositivo móvil que permite realizar la detección de microorganismos presentes en una muestra, se utilizará un campo de aplicación específico a fin de ilustrar de mejor manera las ventajas de la invención, sin embargo, las descripciones basadas en dicho campo de aplicación específico no deben considerarse como una limitante al alcance de la presente invención, descrita en el pliego de reivindicaciones. In the description of the present invention, which corresponds to a mobile device that allows the detection of microorganisms present in a sample, a specific field of application will be used in order to better illustrate the advantages of the invention, however, the descriptions based on said specific field of application should not be considered as a limitation on the scope of the present invention, described in the specification.
El ejemplo específico del campo de aplicación corresponde al aseguramiento de calidad microbiológica en la industria alimentaria, más específicamente, en el monitoreo y detección de microorganismos en una planta de producción animal. Sin embargo, tal como se indicó previamente, el dispositivo de la presente invención puede ser utilizado en otras industrias que requieren monitorear y controlar la calidad microbiológica. Por ejemplo, otras áreas donde podría utilizarse el dispositivo de la presente invención son: aseguramiento de calidad microbiológica del agua, plantas de procesamiento de alimentos, ya sean de origen vegetal o animal, entre otros. The specific example of the field of application corresponds to the assurance of microbiological quality in the food industry, more specifically, in the monitoring and detection of microorganisms in an animal production plant. However, as previously indicated, the device of the present invention can be used in other industries that require monitoring and control of microbiological quality. For example, other areas where the device of the present invention could be used are: microbiological quality assurance of water, food processing plants, whether of vegetable or animal origin, among others.
A fin de evaluar los méritos del dispositivo que se describe en el presente documento, se realizó una búsqueda de documentos relacionados al dispositivo de la presente invención, y a continuación se presenta un breve resumen de cada uno de los documentos encontrados, que tienen alguna relación con el dispositivo de la presente invención. In order to evaluate the merits of the device described in this document, a search of documents related to the device of the present invention was carried out, and a brief summary of each of the documents found, which have some relationship with the device of the present invention.
WO2017025984A1 describe un dispositivo para integrarse a un teléfono inteligente, donde el dispositivo permite la amplificación isotermal de ácidos nucleicos. Se especifica que el teléfono se adapta al dispositivo, de manera que la cámara fotográfica del teléfono puede usarse para registrar mediciones de intensidad en micro-cámaras (micro chambers) donde se lleva a cabo la reacción. WO2017025984A1 describes a device to be integrated into a smartphone, where the device allows isothermal nucleic acid amplification. It is specified that the phone adapts to the device, so that the camera of the phone can be used to record intensity measurements in micro-cameras (micro chambers) where the reaction is carried out.
CN 106010961 A describe un aparato simple para la identificación de patógenos en comida, utilizando como ensayo base amplificación isotermal mediada por bucle (Loop-mediated isothermal amplification, LAMP). Se menciona que el aparato considera el uso de un computador portátil para la colección y procesamiento de información. El aparato considera una placa térmica, donde se realiza la reacción, un dispositivo de excitación fluorescente, un mecanismo de fotografía. CN 106010961 A describes a simple apparatus for the identification of pathogens in food, using as a base test loop-mediated isothermal amplification (Loop-mediated isothermal amplification, LAMP). It is mentioned that the device considers the use of a portable computer for the collection and processing of information. The apparatus considers a thermal plate, where the reaction is carried out, a fluorescent excitation device, a photography mechanism.
US2016231324A1 describe sistemas para detección de alto rendimiento (high-throughput) basados en microfluídica. En las reivindicaciones se menciona que uno de los ensayos que pueden adaptarse al sistema es LAMP. También menciona que el sistema puede diseñarse de manera de adaptarlo para portabilidad, como, por ejemplo, incluyéndolo en una mochila. US2016231324A1 describes high performance throughput systems based on microfluidics. In the claims it is mentioned that one of the tests that can be adapted to the system is LAMP. He also mentions that the system can be designed to adapt it for portability, such as by including it in a backpack.
US2016216210A1 describe un aparato portátil (hardware) para amplificación de ácidos nucleicos, y un software para instalarse en un dispositivo portátil para controlar y obtener los resultados que genera el hardware. En las reivindicaciones se menciona que le método de amplificación de ácidos nucleicos puede ser LAMP. US2016216210A1 describes a portable device (hardware) for nucleic acid amplification, and software to be installed in a portable device to control and obtain the results generated by the hardware. In the claims it is mentioned that the nucleic acid amplification method can be LAMP.
WO2016064894A2 describe un método para detectar virus de ARN, que consiste en una reacción de retro-transcripción acoplada con una reacción de amplificación isotermal. En la memoria descriptiva se menciona que el método puede llevarse a cabo en un dispositivo portátil en un punto de atención (point of care), sin embargo, no entrega detalles de tal dispositivo. WO2016064894A2 describes a method for detecting RNA viruses, which consists of a back-transcription reaction coupled with an isothermal amplification reaction. In the specification it is mentioned that the method can be carried out on a portable device at a point of care, however, it does not provide details of such a device.
US2015141264A1 describe un dispositivo portátil para detección de muestras de ADN específicas. En las reivindicaciones se indica que uno de los sistemas para identificación de ADN que se pueden implementar en el dispositivo es LAMP. No se refiere a identificación de patógenos, ya que el foco de este documento es un sistema para autentificación. US2015141264A1 describes a portable device for detection of specific DNA samples. The claims indicate that one of the systems for DNA identification that can be implemented in the device is LAMP. It does not refer to pathogen identification, since the focus of this document is a system for authentication.
US2014335527A1 describe un dispositivo móvil para análisis de ácidos nucleicos y proteínas. El dispositivo incluye un módulo para extracción de ácidos nucleicos, y además para su amplificación (opcionalmente se indica que puede realizarse usando, entre otros métodos, LAMP). El dispositivo además está dotado de capacidades de comunicación inalámbrica para transferir los resultados. No se encontró referencia a un conjunto de patógenos específicos que podrían detectarse con el sistema. US2014335527A1 describes a mobile device for analysis of nucleic acids and proteins. The device includes a module for nucleic acid extraction, and also for amplification (optionally it is indicated that it can be performed using, among other methods, LAMP). The device is also equipped with wireless communication capabilities to transfer the results. No reference was found to a set of specific pathogens that could be detected with the system.
US2014220667A1 describe un dispositivo portátil para detección de patógenos. Dentro de los métodos que puede implementar el dispositivo, se menciona LAMP. El dispositivo es bastante distinto a lo propuesto en la presente invención, ya que comprende una tira flexible que comprende pocilios donde se van depositando alícuotas de muestra. En el texto se destaca la ventaja de ser un dispositivo útil para uso en POC (point of care). No se menciona la aplicación a patógenos específicos de cerdo. Dentro de los ejemplos de detección se realiza la detección de distintas cepas de E. coli. US2014220667A1 describes a portable device for pathogen detection. Among the methods that the device can implement, LAMP is mentioned. The device is quite different from what is proposed in the present invention, since it comprises a flexible strip comprising wells where aliquots of sample are deposited. The text highlights the advantage of being a useful device for use in POC (point of care). The application to specific pig pathogens is not mentioned. Within the examples of Detection is performed the detection of different strains of E. coli.
CN 102199531 A describe un chip que utiliza microfluídica para realizar detecciones de varias secuencias de ácidos nucleicos en base a una sola muestra. Se indica que se usa LAMP como método de detección. En la última figura muestran un chip de ejemplo de realización para detección de múltiples patógenos en porcinos. No se encontró referencia a un dispositivo móvil para uso en locaciones. CN 102199531 A describes a chip that uses microfluidics to detect several nucleic acid sequences based on a single sample. It is indicated that LAMP is used as the detection method. In the last figure they show an exemplary embodiment chip for detection of multiple pathogens in pigs. No reference was found to a mobile device for location use.
US2010273159A1 describe un método para la identificación de múltiples identidades biológicas en una muestra, que comprende reacciones de amplificación anidadas. En la reivindicación 9 se indica la posibilidad de usar un método de amplificación isotermal, y en la 10 se entregan varias alternativas, donde aparece LAMP. US2010273159A1 describes a method for the identification of multiple biological identities in a sample, comprising nested amplification reactions. In claim 9 the possibility of using an isothermal amplification method is indicated, and in 10 several alternatives are delivered, where LAMP appears.
W02006110855A2 describe métodos para determinar variantes de secuencias. El método descrito usa distintas rondas de amplificación, donde al menos una de ellas se realiza utilizando una amplificación isotermal. En las opciones para amplificación isotermal se menciona LAMP. En el texto del documento se indica que puede aplicarse a muestras provenientes de distintos seres vivos, y se menciona, entre muchas otras opciones, a cerdos. Existen diferencias con la presente invención, pues utiliza soportes móviles en emulsión para inmovilizar algunos ácidos nucleicos. Además, no menciona un dispositivo móvil para llevar a cabo el método. W02006110855A2 describes methods for determining sequence variants. The described method uses different rounds of amplification, where at least one of them is performed using an isothermal amplification. LAMP is mentioned in the options for isothermal amplification. The text of the document indicates that it can be applied to samples from different living beings, and it is mentioned, among many other options, to pigs. There are differences with the present invention, since it uses emulsion movable supports to immobilize some nucleic acids. In addition, it does not mention a mobile device to carry out the method.
EP3187595A1 describe un dispositivo para detectar cepas de Legionella en una muestra. El dispositivo incluye fuentes de calor, fuentes de luz ultra violeta (UV), y detectores ópticos. EP3187595A1 describes a device for detecting Legionella strains in a sample. The device includes heat sources, ultra violet (UV) light sources, and optical detectors.
US2016077006A1 describe un dispositivo que cuenta con fuentes de calor, fuentes de luz, detectores ópticos, donde el dispositivo se puede usar para detectar amplificaciones de ácidos nucleicos. US2016077006A1 describes a device that has heat sources, light sources, optical detectors, where the device can be used to detect nucleic acid amplifications.
US2014349298A1 describe un dispositivo optoeléctrico para detección de patógenos usando amplificación isotermal. Se indica que el dispositivo cuenta con un elemento generador de calor, un diodo emisor de luz UV, un sensor, la lectura se comunica a un dispositivo externo por medio de tecnología inalámbrica. US2014349298A1 describes an optoelectric device for pathogen detection using isothermal amplification. It is indicated that the device has a heat generating element, a UV light emitting diode, a sensor, the reading is communicated to an external device by means of wireless technology.
CN203720089U describe un dispositivo para análisis de muestras usando amplificación isotermal. El dispositivo incluye una fuente de calor para controlar la temperatura de las muestras, una fuente de luz UV para realizar la lectura del resultado, y un sensor de luz UV (sensor óptico). CN203720089U describes a device for sample analysis using isothermal amplification. The device includes a heat source to control the temperature of the samples, a UV light source to read the result, and a UV light sensor (optical sensor).
De todos los documentos encontrados en la búsqueda realizada, ninguno comprende todos y cada uno de los elementos del dispositivo de la presente invención. Of all the documents found in the search performed, none includes all and each of the elements of the device of the present invention.
RESUMEN DE LA INVENCIÓN SUMMARY OF THE INVENTION
La invención corresponde a un dispositivo móvil que permite detectar microorganismos en distintas muestras, donde el dispositivo funciona con base en reacciones de amplificación de ácidos nucleicos. The invention corresponds to a mobile device that allows to detect microorganisms in different samples, where the device works based on nucleic acid amplification reactions.
DESCRIPCIÓN DETALLADA DE LA INVENCIÓN DETAILED DESCRIPTION OF THE INVENTION
La presente invención corresponde a un dispositivo móvil que permite detectar la presencia de múltiples microorganismos predeterminados de manera paralela, donde la detección se realiza por medio del uso de técnicas de amplificación de ácidos nucleicos, combinado con la utilización de sondas específicamente diseñadas para la detección de dichos microorganismos predeterminados. The present invention corresponds to a mobile device that allows the presence of multiple predetermined microorganisms to be detected in parallel, where the detection is carried out through the use of nucleic acid amplification techniques, combined with the use of probes specifically designed for the detection of said predetermined microorganisms.
En un primer aspecto de la invención, el dispositivo comprende al menos los siguientes componentes: i. Fuente de calor controlable, que permita asegurar la mantención de una temperatura predeterminada por un tiempo predeterminado, donde el control de temperatura permite realizar reacciones de amplificación de ácidos nucleicos; ii. Soporte para contenedor de muestras con múltiples cámaras independientes, que permite depositar muestras en cada una de las cámaras; In a first aspect of the invention, the device comprises at least the following components: i. Controllable heat source, which ensures the maintenance of a predetermined temperature for a predetermined time, where the temperature control allows nucleic acid amplification reactions; ii. Support for sample container with multiple independent cameras, which allows depositing samples in each of the cameras;
Ni. Dispositivo de captura de imágenes, que permita obtener una imagen del soporte una vez que se han llevado a cabo las reacciones de amplificación de ácidos nucleicos;  Neither. Image capture device, which allows obtaining an image of the support once the nucleic acid amplification reactions have been carried out;
iv. Fuente de luz UV;  iv. UV light source;
v. Medios de comunicación inalámbrica;  v. Wireless communication media;
vi. Alarmas acústicas y visuales;  saw. Acoustic and visual alarms;
vii. Carcasa; y  vii. Case; Y
viii. Kit de sondas viii Probe kit
En una realización específica, la fuente de calor controlable se selecciona entre: una placa calefactora y una placa tipo peltier. In a specific embodiment, the controllable heat source is selected from: a heating plate and a peltier plate.
En otra realización, el soporte para contenedor de muestras corresponde a una placa perforada, donde cada perforación permite el posicionamiento de cada una de las cámaras presentes en el contenedor de muestras. En una realización particular, el material de la placa perforada se selecciona entre aluminio y cobre. En una realización específica, la placa perforada tiene un tamaño de entre 80 mm y 130 mm por lado, pudiendo tener una forma cuadrada o rectangular. El contenedor de muestras con múltiples cámaras independientes se selecciona entre: placas de 96 pocilios, tiras de 8 tubos, tubos independientes. El volumen de los tubos individuales puede variar entre 20 y 500 pl. In another embodiment, the sample container holder corresponds to a perforated plate, where each perforation allows the positioning of each of the chambers present in the sample container. In a particular embodiment, the perforated plate material is selected from aluminum and copper. In a specific embodiment, the Perforated plate has a size between 80 mm and 130 mm per side, being able to have a square or rectangular shape. The sample container with multiple independent chambers is selected from: 96-well plates, 8-tube strips, independent tubes. The volume of the individual tubes can vary between 20 and 500 pl.
En aún otra realización, el dispositivo de captura de imágenes corresponde a una cámara fotográfica digital. En una realización específica, la cámara permite obtener una imagen de al menos 5 megapíxeles de resolución nativa del sensor. En una realización más específica, el dispositivo de captura de imágenes además puede capturar video con una resolución de al menos 640x480 pixeles a una frecuencia de entre 60 y 90 cuadros por segundo. En otra realización, la captura de video puede realizarse con una resolución de 720p a una frecuencia de 60 cuadros por segundo, y en aun otra realización, la captura de video puede realizarse a 1080p a una frecuencia de 30 cuadros por segundo. In yet another embodiment, the image capture device corresponds to a digital camera. In a specific embodiment, the camera allows to obtain an image of at least 5 megapixels of native resolution of the sensor. In a more specific embodiment, the image capture device can also capture video with a resolution of at least 640x480 pixels at a frequency of between 60 and 90 frames per second. In another embodiment, video capture can be performed with a resolution of 720p at a frequency of 60 frames per second, and in yet another embodiment, video capture can be performed at 1080p at a frequency of 30 frames per second.
En otra realización, la fuente de iluminación se selecciona entre una fuente de iluminación basada en diodos emisores de luz (Light-Emitting diodes, LEDs) blancos, UV, RGB, R, G o B, de tipo LED SMD, COB o de alta potencia, en que la amplitud de onda de la luz emitida por dichos LEDs corresponde a todo el rango visible y UV, es decir, en longitudes de onda de entre 395 a 700 nm. En una realización particular, el circuito LED opera con un voltaje entre 5 y 24 V, siendo la potencia total disipada por éste entre 4 y 20 W. La intensidad luminosa varía entre 70 y 150 mcd por LED. En otra realización, la invención también considera una pantalla LCD con una interfaz táctil, donde se presentan interruptores virtuales o físicos, junto con un regulador de tipo perilla que permite, por ejemplo, navegar por un menú de opciones del dispositivo. En aún otra realización, se considera una pantalla táctil TFT resistiva o capacitiva con interfaz HDMI pequeña de entre 2 y 9 pulgadas. En aún otra realización, la pantalla táctil TFT resistiva o capacitiva utiliza una interfaz SPI entre 2 y 9 pulgadas. In another embodiment, the lighting source is selected from a lighting source based on white light emitting diodes (Light-Emitting diodes, LEDs), UV, RGB, R, G or B, of type SMD, COB or high LED power, in which the wavelength of the light emitted by said LEDs corresponds to the entire visible range and UV, that is, in wavelengths between 395 to 700 nm. In a particular embodiment, the LED circuit operates with a voltage between 5 and 24 V, the total power dissipated by it between 4 and 20 W. The light intensity varies between 70 and 150 mcd per LED. In another embodiment, the invention also considers an LCD screen with a touch interface, where virtual or physical switches are presented, together with a knob-type regulator that allows, for example, to navigate through a menu of device options. In yet another embodiment, a resistive or capacitive TFT touch screen with a small HDMI interface between 2 and 9 inches is considered. In yet another embodiment, the resistive or capacitive TFT touch screen uses a SPI interface between 2 and 9 inches.
En aun otra realización, las alarmas acústicas y visuales corresponden a señales audibles o visibles que se activan, por ejemplo, una vez que el ciclo de detección de microorganismos ha finalizado. In yet another embodiment, the acoustic and visual alarms correspond to audible or visible signals that are activated, for example, once the microorganism detection cycle has ended.
En una realización específica, se consideran indicadores para señalar el encendido del dispositivo con una luz preferentemente verde. La indicación del proceso de calentamiento de la placa perforada se realiza preferentemente con una luz amarilla intermitente entre 5 y 30 min. Finalmente, la señal del final del proceso se realiza preferentemente con una luz verde intermitente y/o una señal acústica también intermitente (de cierta duración). Esta señal visual y/o auditiva indica al usuario que se ha alcanzado la temperatura adecuada para iniciar el proceso de calentamiento de muestras, sugiriéndole al usuario proceder a colocar el contenedor de muestras sobre la placa perforada. In a specific embodiment, indicators are considered to indicate the power of the device with a preferably green light. The indication of the heating process of the perforated plate is preferably performed with an intermittent yellow light between 5 and 30 min. Finally, the signal at the end of the process is preferably performed with an intermittent green light and / or an intermittent acoustic signal (of a certain duration). This visual and / or auditory signal indicates to the user that the appropriate temperature has been reached to start the sample heating process, suggesting the user proceed to place the sample container on the perforated plate.
En una realización particular, el dispositivo de la invención cuenta con un botón de inicio en la carcasa, que una vez presionado por el usuario, o gatillado por un sensor interruptor de fin de carrera, de manera que de forma automática y al colocar el contenedor de muestras se activa el dispositivo y procede a dar inicio a un temporizador para que las muestras se calienten por una determinada cantidad de tiempo, dependiendo de lo que dure la reacción. Una vez que se da inicio a la nueva etapa, el led verde volvería a la normalidad y dejaría de ser intermitente. In a particular embodiment, the device of the invention has a start button on the housing, which once pressed by the user, or triggered by a limit switch sensor, so that automatically and when placing the container of samples the device is activated and proceeds to start a timer so that the samples are heated for a certain amount of time, depending on the duration of the reaction. Once the new stage starts, the green LED would return to normal and cease to be intermittent.
Una vez terminada la etapa anterior, un indicador visual, preferentemente una luz roja y/o una señal acústica, permite sugerir al usuario que el proceso ha terminado y es necesario sacar las muestras del dispositivo. Once the previous stage is finished, a visual indicator, preferably a red light and / or an acoustic signal, allows the user to suggest that the process is over and it is necessary to take the samples from the device.
En otra realización, si durante el proceso se produce un error, una alerta visual, preferentemente una luz roja se activa de forma intermitente. En una realización más específica, el dispositivo cuenta además con un botón o interfase que permite restablecer el proceso para iniciarlo nuevamente, o en la alternativa, simplemente basta con prender y apagar el equipo. In another embodiment, if an error occurs during the process, a visual alert, preferably a red light is activated intermittently. In a more specific embodiment, the device also has a button or interface that allows to reset the process to start it again, or in the alternative, simply turn the equipment on and off.
En otra realización, los medios de comunicación inalámbrica son seleccionados entre: Wi- Fi, 3G, LoRa. En una realización específica, los medios de comunicación inalámbrica permiten enviar la información resultante de utilizar el dispositivo de la presente invención, es decir, la captura de imágenes o video. En una realización aún más específica, la información de imágenes o video es enviada a un servidor remoto (máquina virtual) en el cual se analiza la información obtenida en las imágenes o vídeo. En particular, este análisis consiste en la segmentación de la imagen total en unidades singulares. Cada unidad singular corresponde a una réplica de la muestra tomada y se compara, mediante un algoritmo clasificador, contra al menos dos conjuntos de imágenes curadas manualmente para determinar a qué conjunto pertenecería cada segmento problema, lo que permite saber si la detección fue positiva o negativa en la muestra correspondiente a ese segmento individual. Una vez clasificados todos los segmentos individuales, se ejecuta una automatización que almacena los datos y genera un reporte para el usuario final. In another embodiment, the wireless communication media is selected from: Wi-Fi, 3G, LoRa. In a specific embodiment, the wireless communication means allows the information resulting from using the device of the present invention to be sent, that is, the capture of images or video. In an even more specific embodiment, the image or video information is sent to a remote server (virtual machine) in which the information obtained in the images or video is analyzed. In particular, this analysis consists of the segmentation of the total image into singular units. Each singular unit corresponds to a replica of the sample taken and is compared, using a classifying algorithm, against at least two sets of manually curated images to determine which set each problem segment would belong to, which allows to know if the detection was positive or negative in the sample corresponding to that individual segment. Once all the individual segments have been classified, an automation is executed that stores the data and generates a report for the end user.
Finalmente, los datos procesados en el computador son desplegados mediante alguna de las siguientes formas: a través de una aplicación web, a una aplicación presente en un dispositivo móvil, como por ejemplo un teléfono inteligente, entre otros. EJEMPLOS Finally, the data processed on the computer is displayed in one of the following ways: through a web application, to an application present on a mobile device, such as a smartphone, among others. EXAMPLES
Ejemplo 1 : Dispositivo conectado a la nube por wifi para medir ácidos nucleicos. Example 1: Device connected to the cloud by Wi-Fi to measure nucleic acids.
Este dispositivo permite diagnosticar la presencia de múltiples patógenos en minutos (20- 60 min). El dispositivo consta de una placa calefactora de cobre y fibra de vidrio perforada de 150 x 100 mm que alcanza temperaturas de hasta 100°C, un transformador fuente de poder de 12 V 8 Amp, una cámara de 5 megapíxeles de resolución nativa del sensor con capacidad de 2592 x 1944 píxeles, además es compatible con video de 1080p30, 720p60 y 640x480p60/90, una fuente de iluminación diseñada a partir de LEDs UV de 12 V, 12 W de potencia por metro y longitud de onda de 395 - 405nm, capacidad inalámbrica a 2.4Ghz con un alcance de 60 metros en lugares cerrados y hasta 1200 metros en lugares exteriores. Una placa metálica de aluminio perforada de 125 x 83 mm y 10 mm de alto para posicionar una placa microtest de poliestireno con 96 pocilios de 0.1 pL . Alarma acústica y luminosa por fin de ciclo. Funciona con un kit asociado que contiene sondas para detección específica de microorganismos seleccionados mediante amplificación isotermal de DNA. This device allows diagnosing the presence of multiple pathogens in minutes (20-60 min). The device consists of a 150 x 100 mm perforated copper and fiberglass heating plate that reaches temperatures of up to 100 ° C, a 12 V 8 Amp power source transformer, a 5 megapixel camera native resolution of the sensor with 2592 x 1944 pixels capacity, it is also compatible with 1080p30, 720p60 and 640x480p60 / 90 video, a lighting source designed from 12V UV LEDs, 12 W of power per meter and wavelength of 395 - 405nm, 2.4Ghz wireless capacity with a range of 60 meters indoors and up to 1200 meters outdoors. A perforated aluminum metal plate 125 x 83 mm and 10 mm high to position a polystyrene microtest plate with 96 wells of 0.1 pL. Acoustic and luminous alarm at the end of the cycle. It works with an associated kit that contains probes for specific detection of microorganisms selected by isothermal DNA amplification.
Para realizar el análisis, el usuario deposita la muestra en cada uno de los pocilios de detección. La placa se calienta y se realiza la reacción de amplificación isotermal. Al cabo de 20-60 min (a 60°C), se ilumina la placa con luz UV y se realiza una captura de imagen. La imagen es enviada vía wifi a un computador remoto para análisis y almacenamiento en una base de datos. El análisis se realiza mediante un algoritmo de segmentación de imagen y un algoritmo de clasificación en base a colorimetría para determinar si el resultado de cada reacción individual es positivo o negativo. Los datos una vez procesados se cargan en una aplicación web y una móvil para la visualización de los resultados y la entrega de un reporte al usuario. To perform the analysis, the user deposits the sample in each of the detection wells. The plate is heated and the isothermal amplification reaction is performed. After 20-60 min (at 60 ° C), the plate is illuminated with UV light and an image capture is performed. The image is sent via Wi-Fi to a remote computer for analysis and storage in a database. The analysis is performed using an image segmentation algorithm and a classification algorithm based on colorimetry to determine whether the result of each individual reaction is positive or negative. Once the data is processed, it is uploaded to a web application and a mobile application for viewing the results and delivering a report to the user.
Ejemplo 2: Detección de Staphylococcus aureus desde muestras nasales de porcinos Example 2: Detection of Staphylococcus aureus from swine nasal samples
Para la detección de Staphylococcus aureus, se toman muestras desde los orificios nasales de los porcinos usando una tórula estéril, la cual se conserva, en un tubo Eppendorf con un reactivo estabilizador de DNA (DNA Shield). Las muestras se mezclan con una solución de extracción de DNA rápido (20 min), mediante un protocolo de agitación y calor. Posteriormente, las muestras se agregan en una placa de 96 pocilios, que ya contiene los reactivos correspondientes el kit para detectar este patógeno mediante amplificación isotermal de ADN (enzima, dNTPS, partidores, buffer y marcador fluorescente o colorimétrico). El protocolo para agregar las muestras a la placa consiste en cargar 10 pL desde la posición D4 hasta G9, llenando en total 36 pocilios de la placa de 96 pocilios (ver Fig. 1 , placa 96). La medición se realiza por triplicado, permitiendo medir 12 muestras al mismo tiempo (12 porcinos diferentes). Los pocilios restantes se utilizan como controles de la reacción (positivo y negativo). La reacción ocurre a 60°C por 20-40 minutos y el cambio de color es un indicador positivo de la reacción. El límite de detección es de 10 fentogramos/ul de concentración de ADN. La detección de Staphylococcus aureus mediante este dispositivo desde muestras nasales de porcinos requiere entre 40-60 minutos incluyendo el procesamiento de la muestra y la reacción de amplificación de ADN. For the detection of Staphylococcus aureus, samples are taken from the porcine nostrils using a sterile torula, which is preserved, in an Eppendorf tube with a DNA stabilizing reagent (DNA Shield). The samples are mixed with a rapid DNA extraction solution (20 min), using a stirring and heat protocol. Subsequently, the samples are added in a 96-well plate, which already contains the corresponding reagents the kit to detect this pathogen by isothermal DNA amplification (enzyme, dNTPS, splitters, buffer and fluorescent or colorimetric marker). The protocol for adding the samples to the plate consists of loading 10 pL from position D4 to G9, filling in total 36 wells of the 96-well plate (see Fig. 1, plate 96). The measurement is done in triplicate, allowing 12 samples to be measured at the same time (12 different pigs). The remaining wells are used as reaction controls (positive and negative). The reaction occurs at 60 ° C for 20-40 minutes and the color change is a positive indicator of the reaction. The detection limit is 10 fentograms / ul of DNA concentration. The detection of Staphylococcus aureus by this device from porcine nasal samples requires between 40-60 minutes including sample processing and DNA amplification reaction.
APLICACIÓN INDUSTRIAL INDUSTRIAL APPLICATION
La presente invención tiene aplicación en toda industria que requiera monitorear o determinar la presencia de microorganismos específicos en distintos tipos de muestras. Una industria específica, por ejemplo, corresponde a la industria de producción de animales para consumo humano. Una aplicación específica es en la detección de patógenos que producen enfermedades respiratorias en cerdos de producción. The present invention has application in any industry that requires monitoring or determining the presence of specific microorganisms in different types of samples. A specific industry, for example, corresponds to the animal production industry for human consumption. A specific application is in the detection of pathogens that produce respiratory diseases in production pigs.

Claims

REIVINDICACIONES
1. Dispositivo móvil que permite detectar la presencia de múltiples microorganismos predeterminados de manera paralela, donde la detección se realiza por medio del uso de técnicas de amplificación de ácidos nucleicos, combinado con la utilización de sondas específicamente diseñadas para la detección de dichos microorganismos predeterminados, CARACTERIZADO porque comprende al menos los siguientes componentes: 1. Mobile device that allows to detect the presence of multiple predetermined microorganisms in parallel, where the detection is carried out through the use of nucleic acid amplification techniques, combined with the use of probes specifically designed for the detection of said predetermined microorganisms, CHARACTERIZED because it comprises at least the following components:
i. Fuente de calor controlable, que permita asegurar la mantención de una temperatura predeterminada por un tiempo predeterminado, donde el control de temperatura permite realizar reacciones de amplificación de ácidos nucleicos;  i. Controllable heat source, which ensures the maintenance of a predetermined temperature for a predetermined time, where the temperature control allows nucleic acid amplification reactions;
¡i. Soporte para contenedor de muestras con múltiples cámaras independientes, que permite depositar muestras en cada una de las cámaras;  I. Support for sample container with multiple independent cameras, which allows depositing samples in each of the cameras;
Ni. Dispositivo de captura de imágenes, que permita obtener una imagen del soporte una vez que se han llevado a cabo las reacciones de amplificación de ácidos nucleicos;  Neither. Image capture device, which allows obtaining an image of the support once the nucleic acid amplification reactions have been carried out;
iv. Fuente de luz UV;  iv. UV light source;
v. Medios de comunicación inalámbrica;  v. Wireless communication media;
vi. Alarmas acústicas y visuales;  saw. Acoustic and visual alarms;
vii. Carcasa; y  vii. Case; Y
viii. Kit de sondas.  viii Probe kit
2. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque la fuente de calor controlable se selecciona entre: placa calefactora y placa tipo peltier. 2. Mobile device according to claim 1, CHARACTERIZED in that the controllable heat source is selected from: heating plate and peltier plate.
3. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque el soporte para contenedor de muestras corresponde a una placa perforada, donde cada perforación permite el posicionamiento de cada una de las cámaras presentes en el contenedor de muestras. 3. Mobile device according to claim 1, CHARACTERIZED because the holder for sample container corresponds to a perforated plate, where each perforation allows the positioning of each of the chambers present in the sample container.
4. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque el material de la placa perforada se selecciona entre aluminio y cobre. 4. Mobile device according to claim 1, CHARACTERIZED in that the perforated plate material is selected from aluminum and copper.
5. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque la placa perforada tiene un tamaño de entre 80 mm y 130 mm por lado, pudiendo tener una forma cuadrada o rectangular. 5. Mobile device according to claim 1, CHARACTERIZED because the perforated plate has a size between 80 mm and 130 mm per side, and can have a square or rectangular shape.
6. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque el contenedor de muestras con múltiples cámaras independientes se selecciona entre: placas de 96 pocilios, tiras de 8 tubos, tubos independientes. 6. Mobile device according to claim 1, CHARACTERIZED in that the sample container with multiple independent chambers is selected from: 96-well plates, 8-tube strips, independent tubes.
7. Dispositivo móvil de acuerdo a la reivindicación 6, CARACTERIZADO porque el volumen de los tubos individuales puede variar entre 20 y 500 pl. 7. Mobile device according to claim 6, CHARACTERIZED because the volume of the individual tubes can vary between 20 and 500 pl.
8. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque el dispositivo de captura de imágenes corresponde a una cámara fotográfica digital. 8. Mobile device according to claim 1, CHARACTERIZED in that the image capture device corresponds to a digital camera.
9. Dispositivo móvil de acuerdo a la reivindicación 8, CARACTERIZADO porque la cámara permite obtener una imagen de al menos 5 megapíxeles de resolución nativa del sensor. 9. Mobile device according to claim 8, CHARACTERIZED in that the camera allows to obtain an image of at least 5 megapixels of native resolution of the sensor.
10. Dispositivo móvil de acuerdo a la reivindicación 8, CARACTERIZADO porque el dispositivo de captura de imágenes además puede capturar video con una resolución de al menos 640x480 pixeles a una frecuencia de entre 60 y 90 cuadros por segundo, o la captura de video puede realizarse con una resolución de 720p a una frecuencia de 60 cuadros por segundo, o la captura de video puede realizarse a 1080p a una frecuencia de 30 cuadros por segundo. 10. Mobile device according to claim 8, CHARACTERIZED because the image capture device can also capture video with a resolution of at least 640x480 pixels at a frequency of between 60 and 90 frames per second, or video capture can be performed with a resolution of 720p at a frequency of 60 frames per second, or video capture can be performed at 1080p at a frequency of 30 frames per second.
1 1 . Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque la fuente de iluminación se selecciona entre una fuente de iluminación basada en diodos emisores de luz (Light-Emitting diodes, LEDs) blancos, UV, RGB, R, G o B, de tipo LED SMD, COB o de alta potencia, en que la amplitud de onda de la luz emitida por dichos LEDs corresponde a todo el rango visible y UV, es decir, en longitudes de onda de entre 395 a 700 nm. eleven . Mobile device according to claim 1, CHARACTERIZED in that the lighting source is selected from a lighting source based on white light emitting diodes (Light-Emitting diodes, LEDs), UV, RGB, R, G or B, of type SMD, COB or high power LEDs, in which the wavelength of the light emitted by said LEDs corresponds to the entire visible range and UV, that is, in wavelengths between 395 to 700 nm.
12. Dispositivo móvil de acuerdo a la reivindicación 11 , CARACTERIZADO porque el circuito LED opera con un voltaje entre 5 y 24 V, siendo la potencia total disipada por éste entre 4 y 20 W. La intensidad luminosa varía entre 70 y 150 mcd por LED. 12. Mobile device according to claim 11, CHARACTERIZED because the LED circuit operates with a voltage between 5 and 24 V, the total power dissipated by it between 4 and 20 W. The light intensity varies between 70 and 150 mcd per LED. .
13. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque además comprende una pantalla LCD con una interfaz táctil, donde se presentan interruptores virtuales o físicos, junto con un regulador de tipo perilla que permite, por ejemplo, navegar por un menú de opciones del dispositivo. 13. Mobile device according to claim 1, CHARACTERIZED in that it further comprises an LCD screen with a touch interface, where virtual or physical switches are presented, together with a knob-type regulator that allows, for example, to navigate through a menu of options Of the device.
14. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque además comprende una pantalla táctil TFT resistiva o capacitiva con interfaz HDMI pequeña de entre 2 y 9 pulgadas, donde la pantalla táctil TFT resistiva o capacitiva utiliza una interfaz SPI entre 2 y 9 pulgadas. 14. Mobile device according to claim 1, CHARACTERIZED in that it further comprises a resistive or capacitive TFT touch screen with a small HDMI interface between 2 and 9 inches, where the resistive or capacitive TFT touch screen uses a SPI interface between 2 and 9 inches .
15. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque las alarmas acústicas y visuales corresponden a señales audibles o visibles que se activan, por ejemplo, una vez que el ciclo de detección de microorganismos ha finalizado. 15. Mobile device according to claim 1, CHARACTERIZED because the acoustic and visual alarms correspond to audible or visible signals that are activated, for example, once the microorganism detection cycle has ended.
16. Dispositivo móvil de acuerdo a la reivindicación 1 , CARACTERIZADO porque los medios de comunicación inalámbrica son seleccionados entre: Wi-Fi, 3G, LoRa. 16. Mobile device according to claim 1, CHARACTERIZED because the wireless communication means are selected from: Wi-Fi, 3G, LoRa.
17. Dispositivo móvil de acuerdo a la reivindicación 16, CARACTERIZADO porque los medios de comunicación inalámbrica permiten enviar la información resultante de utilizar el dispositivo de la presente invención, es decir, la captura de imágenes o video, a un servidor remoto (máquina virtual) en el cual se analiza la información obtenida en las imágenes o vídeo. 17. Mobile device according to claim 16, CHARACTERIZED in that the wireless communication means allow sending the information resulting from using the device of the present invention, that is, the capture of images or video, to a remote server (virtual machine) in which the information obtained in the images or video is analyzed.
18. Método de análisis para procesar los datos producidos por un dispositivo móvil descrito en las reivindicaciones 1 a 17, CARACTERIZADO porque el análisis consiste en la segmentación de la imagen total en unidades singulares, donde cada unidad singular corresponde a una réplica de la muestra tomada y se compara, mediante un algoritmo clasificador, contra al menos dos conjuntos de imágenes curadas manualmente para determinar a qué conjunto pertenecería cada segmento problema, lo que permite saber si la detección fue positiva o negativa en la muestra correspondiente a ese segmento individual, donde una vez clasificados todos los segmentos individuales, se ejecuta una automatización que almacena los datos y genera un reporte para el usuario final. 18. Method of analysis for processing the data produced by a mobile device described in claims 1 to 17, CHARACTERIZED because the analysis consists in the segmentation of the total image into singular units, where each singular unit corresponds to a replica of the sample taken and it is compared, by means of a classifying algorithm, against at least two sets of manually curated images to determine to which set each problem segment would belong, which allows to know if the detection was positive or negative in the sample corresponding to that individual segment, where a Once all the individual segments have been classified, an automation is executed that stores the data and generates a report for the end user.
19. Método de acuerdo a la reivindicación 18, CARACTERIZADO porque los datos procesados en el computador son desplegados mediante alguna de las siguientes formas: a través de una aplicación web, a una aplicación presente en un dispositivo móvil, como por ejemplo un teléfono inteligente, entre otros. 19. Method according to claim 18, CHARACTERIZED because the data processed in the computer is displayed in any of the following ways: through a web application, to an application present in a mobile device, such as a smartphone, among others.
PCT/CL2019/050022 2018-03-28 2019-03-27 Mobile device that allows the detection of determined microorganisms in different types of samples WO2019183739A1 (en)

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