A system and a method for point-of-care detection of a bacterial infection

Abstract

The present disclosure relates to a system (100) for the detection of bacterial infection in humans. The system comprises an outer surface (101), a pressure-sensitive polyurethane film (103), a microfluidic channel (105), at least one marker (106) and an outlet for removing an excess biofluid (102). The system optionally comprises a lateral flow test strip (200). The system is safe, uses specific antigen-antibody interaction, is accurate, non-invasive allows rapid detection / screening of tuberculosis patients, and is easy to handle without health workers. Further, the present disclosure relates to a method for point-of-care detection of bacterial infection with or without a lateral flow strip (200). The method is specific, non-invasive, rapid, simple, has greater patient compliance and requires less attention from a health care assistant. The system and the method are specifically used to detect Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus.

Description

[0001] A SYSTEM AND A METHOD FOR POINT-OF-CARE DETECTION OF A BACTERIAL INFECTION

[0002] TECHNICAL FIELD

[0003] The present disclosure relates to a system and a method for point-of-care detection of a bacterial infection. The present disclosure relates particularly, but not exclusively, to the system and the method for point-of-care detection of Tuberculosis from a biofluid, such as sweat. The system of the present disclosure incorporates a microfluidic system and optionally a lateral flow strip in a single unit.

[0004] DEFINITIONS

[0005] As used in the present disclosure, the following terms are generally intended to have the meaning as set forth below, except to the extent that the context in which they are used indicates otherwise.

[0006] Point-of-care (POC): The term “Point-of-care” refers to medical testing that is performed outside of a laboratory setting. POC testing is also known as bedside testing, near-patient testing, remote testing, mobile testing and rapid diagnostics. In general, the term can encompass any patient medical test that is given ad hoc and provides quick results.

[0007] Biofluid or biological fluid: The term “Biofluid” refers to any bio-organic fluid produced by an organism such as sweat, saliva, intestinal fluid, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid.

[0008] Biocompatibility: The term “Biocompatibility” refers to a general term describing the property of material being compatible with living tissue. Biocompatible materials do not produce a toxic or immunological response when exposed to the body or bodily fluids.

[0009] Microfluidics: The term “microfluidics” refers to the study of systems that can process small quantities of fluids by using tiny channels having dimensions at the microscale.

[0010] BACKGROUND

[0011] The background information herein below relates to the present disclosure but is not necessarily prior art.

[0012] Infectious diseases are widespread in the world. One of the most prominent and deadlier ones is Tuberculosis which is caused by Mycobacterium tuberculosis (MTB). It is usually characterized by the production of antibodies to specific disease-related antigens and the formation of granulomas in the affected organ. It commonly affects the lungs. Common symptoms include cough, which lasts for longer than two weeks. Weight loss, low-grade evening fevers, and excessive night sweats are also common among the patients. Cough is associated with phlegm production and blood. According to the World Health Organization (WHO), the infectious disease tuberculosis kills about three million people every year. Globally, there are millions of people who suffer from tuberculosis. Millions die every year due to this disease. In developing countries, the disease is rampant. People living in poverty-stricken areas bear the greatest burden of the disease. Most of those affected are in the economically productive age group. Even though it is a bacterial infection, there are significant associations between socioeconomic conditions and tuberculosis.

[0013] There are several conventional tests for the detection of MTB, however, these conventional tests all have disadvantages.

[0014] One conventional test is a purified protein derivative (PPD) skin test. In this test, a PPD is injected subcutaneously into a patient, and after 2 to 3 days, the site of injection is observed to determine whether there is hardening and / or discolouration of the patient’s skin. This test is believed to be only about 50-75 % accurate. In persons that have been immunized with Mycobacterium Bovis BCG, there is a high percentage of false positives. Another disadvantage of the PPD skin test is that it requires a needle injection, and patient compliance is low in coming back for observation by trained medical personnel for hardening and / or discolouration. This test is typically used as a screening test for people who do not have symptoms of tuberculosis. Another conventional test is a culture test. However, this test takes up to 6 to 8 weeks since that is how long it takes for the MTB organism to grow to a visibly detectable amount. Another disadvantage of the culture test is that typically only about half of all samples wherein the Mycobacterium tuberculosis is present grow to the point where it is detectable. Thus, this method has about 50% false negatives.

[0015] Another conventional test is a chest x-ray. However, this method does not work if the person undergoing the chest x-ray has been recently infected with Mycobacterium tuberculosis . Another disadvantage is it requires attendance at health centres several times before a diagnosis is made. The diagnosis is also challenging in children, pregnant women who cannot be exposed to radiation, and people living in remote areas without easy access to healthcare services.

[0016] Another conventional test is the presence of acid-fast bacilli (AFB) in sputum smears. This requires the collection of multiple sputum samples. The new diagnostic method, based on sputum molecular testing, is expensive, requires some technological expertise, and has lots of infrastructure support which is not readily available in rural areas. These factors contribute to delays in diagnosis.

[0017] Further, there are other point-of-care tests (POCT) available for childhood tuberculosis such as the detection of Lipoarabinomannan (LAM) in urine using lateral flow assays, loop-mediated isothermal amplification (LAMP), PCR-based point-of-care tests and point-of-care ultrasound (POCUS) imaging for pulmonary tuberculosis, however, aforestated conventional tests and other point-of-care tests for childhood tuberculosis have are not up to the benchmark and have their own disadvantage such as high percentage of false positives, requires needle injection, poor patient compliance, takes longer time for detection, less specific, low sensitivity, expensive, complex in nature, requires attendance at health centres for several times before diagnosis, requires some technological expertise, and lots of infrastructure support and the like.

[0018] Therefore, in light of the foregoing discussion, there exists a need to provide a system and a method for point-of-care detection of bacterial infection that mitigates the drawbacks mentioned hereinabove or at least provides a useful alternative. OBJECTS

[0019] Some of the objects of the present disclosure, which at least one embodiment herein satisfies, are as follows.

[0020] An object of the present disclosure is to ameliorate one or more problems of the prior art orto at least provide a useful alternative.

[0021] An object of the present disclosure is to provide a system for the detection of bacterial infection. Another object of the present disclosure is to provide a system for the detection of Tuberculosis in a subject, particularly in humans.

[0022] Another object of the present disclosure is to provide a system that is safe, accurate and easy to handle without health workers.

[0023] Still, another object of the present disclosure is to provide a system that is non-invasive and allows rapid detection / screening of tuberculosis patients in real-time.

[0024] Yet another object of the present disclosure is to provide a system that uses specific antigenantibody interaction.

[0025] Still, another object of the present disclosure is to provide a method for point-of-care detection of Tuberculosis.

[0026] Yet another object of the present disclosure is to provide a method for point-of-care detection of Tuberculosis which is simple, accurate, specific, sensitive, patient-centric, quick, requires no assistance from health workers, and provides real-time qualitative and qualitative data for further monitoring.

[0027] Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of the present disclosure.

[0028] SUMMARY

[0029] The present disclosure relates to a system and a method for point-of-care detection of bacterial infections, such as Tuberculosis. The present disclosure relates particularly, but not exclusively, to the system and the method for point-of-care detection of Tuberculosis from the biofluid, such as sweat.

[0030] The present disclosure provides a solution to the existing problem of how bacterial infection is specifically, without assistance from health workers and quickly detected non-invasively using biofluid. The aim of the present disclosure is to provide a solution that overcomes the problems encountered in the prior art and provides a system and a method for point-of-care detection of tuberculosis.

[0031] One or more objectives of the present disclosure are achieved by the solutions provided in the enclosed independent claims. Advantageous implementations of the present disclosure are further defined in the dependent claims.

[0032] In one aspect, the present disclosure provides a system (100) for the detection of bacterial infection in the subject, the system (100) comprises a biofluid collector which consists of an outer surface (101) that is made up of biocompatible material for contacting a skin surface of the subject and a pressure -sensitive polyurethane film (103), a microfluidic channel (105), at least one marker (106), optionally, a lateral flow test strip (200), an outlet for removing an excess biofluid (102).

[0033] The outer surface comprises an inlet (104) that receives the biofluid excreted from the skin of the subject.

[0034] The lateral flow test strip (200) optionally is connected with the biofluid collector and microfluidic channel through the outlet (102).

[0035] The microfluidic channel (105) is in fluid communication with the biofluid collector, the marker (105) and the lateral flow test strip (200).

[0036] The parts of the system (100) are arranged from one to other to form a single unit.

[0037] In another implementation form, the bacterial infection is at least one selected from the group consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus.

[0038] In another implementation form, the bacterial infection is Tuberculosis.

[0039] In another implementation form, the system (100) detects Tuberculosis, diagnose and monitors the infection in real-time.

[0040] In another implementation form, the system (100) detects active and latent stages of Tuberculosis.

[0041] In another implementation form, the subject is a human.

[0042] In another implementation form, the system (100) is a detachable device that is attached directly to the skin surface and the device has a length ranging from 5 to 10 cm and a width ranging from 0.5 cm to 2 cm.

[0043] In another implementation form, the device is attached to the skin surface through a skin-safe adhesive selected from marker adhesive and paper adhesive.

[0044] In another implementation form, the biofluid is at least one selected from the group consisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid.

[0045] In another implementation form, the biofluid is sweat.

[0046] In another implementation form, the biofluid is saliva.

[0047] In another implementation form, the biocompatible material of the outer surface (101) is a hydrophilic foam.

[0048] In another implementation form, the hydrophilic foam is highly permeable to the skin surface. In another implementation form, the outer surface (101) covers an external surface area in the range of 20 mm2to 25 mm2.

[0049] In another implementation form, the inlet (104) consists of pores that direct the flow of sweat and dispense it to the microfluidic channel (105) followed by optionally the lateral flow test strip (200).

[0050] In another implementation form, the pores are characterized by having an average pore size of less than 0.1 mm.

[0051] In another implementation form, the microfluidic channel (105) has an internal volume to trap the sweat ranging from 50 ul to 100 ul in a time period ranging from 5 to 10 minutes.

[0052] In another implementation form, the system (100) further comprises a sensor (107) which is embedded in the microfluidic channel (105).

[0053] In another implementation form, the sensor is a chemical sensor.

[0054] The chemical sensor detects the analyte of interest. The chemical sensor reacts with the biomarker in the sweat and produces a colourimetric / colour change if the biomarker is present. So the chemical sensor varies depending on the presence of the biomarker used and the intensity of the colour change is determined by the level of the biomarkers in the sweat.

[0055] The chemical sensor has specificity in the range of 80 to 90%, preferably 88% and the sensitivity in the range of 90 to 95%, preferably 92%.

[0056] In another implementation form, the biofluid in the microfluidic channel (105) has a unidirectional flow.

[0057] In another implementation form, the marker (106) is human serum amyloid protein AA2.

[0058] In another implementation form, the marker (106) is embedded into the microfluidic channel (105). The marker (106) is embedded into the microfluidic channel (105) for early detection of Tuberculosis directly from the biofluid.

[0059] In another implementation form, the lateral flow test strip (200) comprises a specimen pad (203) having dispensed sweat, a conjugate pad (204) comprising at least one detector molecule, and a test line (199) comprising capture molecules.

[0060] In another implementation form, the lateral flow test strip (200) detects the presence of the specific marker / protein associated with Mycobacterium tuberculosis .

[0061] In another implementation form, the lateral flow test strip is 100% specific for detecting Mycobacterium tuberculosis . In another implementation form, each of the detector molecules comprises a first antigen-specific antibody conjugated to a detectable particle.

[0062] In another implementation form, the detectable particle is gold nanoparticles.

[0063] In another implementation form, the gold nanoparticles have a size in the range from 3 nm to 40 nm.

[0064] In another implementation form, the detector molecules comprise one or more first antigenspecific antibodies conjugated to the detectable particle.

[0065] In another implementation form, the first antigen-specific antibody is specific for Mycobacterium tuberculosis .

[0066] In another implementation form, the capture molecules comprise a second antigen-specific antibody.

[0067] In another implementation form, the second antigen-specific antibody is specific for Mycobacterium tuberculosis .

[0068] In another implementation form, the specimen pad contains a pretreated buffer and / or is added by a dropper.

[0069] In another implementation form, the buffer is a SAA assay buffer.

[0070] In another implementation form, the outlet (102) has a diameter in the range of 2 to 10 mm.

[0071] In another implementation form, the system uses capillary motion to control the flow of sweat among its all components.

[0072] In another implementation form, the system provides test results in about 30 minutes or less.

[0073] In another implementation form, the system provides test results in about 20 minutes or less.

[0074] In another implementation form, the system provides test results in about 10 minutes.

[0075] In another implementation form, the system provides test results with an accuracy of about 80% or greater.

[0076] In another implementation form, the system provides test results with an accuracy of about 85% or greater.

[0077] In another implementation form, the system provides test results with an accuracy of about 88% or greater. In another implementation form, the microfluidic channel (105) is further embedded with latex particles (108) for the detection of Mycobacterium tuberculosis .

[0078] In another implementation form, the system (100), further comprises (a) a processing unit, which receives an output signal of the sensor and detects said signal qualitatively and quantitatively and (b) a transmitter, which receives an output signal of the sensor and transmits said signal to a remote server and / or to a portable electronic device.

[0079] In another implementation form, the remote server and the portable electronic device comprise an algorithm, which generates qualitative and quantitative data for further diagnosing and / or monitoring.

[0080] In another implementation form, the system (100) comprises an indicator, wherein the processing unit and the transmitter are configured to transmit a positive indication signal or a negative indication signal to the indicator, thereby displaying the outcome.

[0081] In another implementation form, the processing unit, the remote server and the portable electronic device further comprise an algorithm configured to differentiate between the response of the sensor to the biofluid excreted from the skin and the response of the sensor to a stimulus selected from the group consisting of temperature, humidity, lateral strain, and combinations thereof.

[0082] In another implementation form, the system generates real-time quantitative and qualitative data for early detection, diagnosis and / or monitoring of Mycobacterium tuberculosis in the subject.

[0083] In another aspect, the present disclosure provides a method for point-of-care detection of a bacterial infection.

[0084] The method comprises the following steps: a) obtaining a biofluid excreted from the skin of the subject by placing the system (100) of claim 1 on the skin surface of the subject for 10 min to collect the biofluid; b) dispensing the collected biofluid to a microfluidic channel (105) through an inlet (104); and c) allowing the dispensed biofluid in the microfluidic channel (105) for 15 to 30 minutes followed by detecting the presence and absence of the bacterial infection in the subject.

[0085] In another implementation form, the bacterial infection is at least one selected from the group consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus.

[0086] In another implementation form, the bacterial infection is Tuberculosis.

[0087] In another implementation form, the method detects Tuberculosis, diagnose and monitors the infection in real-time. In another implementation form, the method detects active and latent stages of Tuberculosis.

[0088] In another implementation form, the subject is human.

[0089] In another implementation form, the biofluid is at least one selected from the group consisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid.

[0090] In another implementation form, the biofluid is sweat.

[0091] In another implementation form, the biofluid is saliva.

[0092] In another implementation form, the biofluid is dispensed from the inlet (104) to the microfluidic channel (105) and other components through a capillary motion.

[0093] In another implementation form, the microfluidic channel (105) comprising a chemical sensor (107) wherein the chemical sensor (107) is exposed to the biofluid followed by measuring an output signal of the chemical sensor upon exposure to the biofluid.

[0094] In another implementation form, the formation of double lines (109, 201) confirms the presence of Mycobacterium tuberculosis in the subject (fig. 5(a) and 6(a)) and wherein the formation of a single line (110, 202) confirms the absence of Mycobacterium tuberculosis in the subject (fig. 5(b) and 6 (b)).

[0095] In another implementation form, the formation of a single test line (203) confirms the presence of an indeterminate result in the subject (fig. 5c).

[0096] In another implementation form, the system (100) is placed on the chest, hands, legs and inner arm skin of the subject.

[0097] In another implementation form, a lateral flow strip (200) is optionally attached to the system (100) through the outlet (102).

[0098] In another implementation form, the method provides test results in about 30 minutes or less.

[0099] In another implementation form, the method provides test results in about 20 minutes or less.

[0100] In another implementation form, the method provides test results in about 10 minutes.

[0101] In another implementation form, the method provides test results with an accuracy of about 80% or greater.

[0102] In another implementation form, the method provides test results with an accuracy of about 85% or greater. In another implementation form, the method provides test results with an accuracy of about 88% or greater.

[0103] It is to be appreciated that all the aforementioned implementation forms can be combined.lt is noted that all devices, elements, circuitry, units, and means described in the present disclosure could be implemented in the software or hardware elements or any kind of combination thereof. All steps which are performed by the various entities described in the present disclosure, as well as the functionalities described to be performed by the various entities, are intended to mean that the respective entity is adapted to or configured to perform the respective steps and functionalities. Even if, in the following description of specific embodiments, a specific functionality or step to be performed by external entitiesis not reflected in the description of a specific detailed element of that entity that performs that specific step or functionality, it should be clear for a skilled person that these methods and functionalities can be implemented in respective software or hardware elements, or any kind of combination thereof. It will be appreciated that features of the present disclosure are susceptible to being combined in various combinations without departing from the scope of the present disclosure as defined by the appended claims.

[0104] Additional aspects, advantages, features, and objects of the present disclosure would be made apparent from the drawings and the detailed description of the illustrative implementations construed in conjunction with the appended claims that follow.

[0105] BRIEF DESCRIPTION OF THE DRAWING

[0106] The summary above, as well as the following detailed description of illustrative embodiments, is better understood when read in conjunction with the appended drawing. For the purpose of illustrating the present disclosure, exemplary constructions of thedisclosure are shown in the drawing. However, the present disclosure is not limited to specific methods and instrumentalities disclosed herein. Moreover, those skilled in the art will understand that the drawings are not to scale. Wherever possible, like elements have been indicated by identical numbers.

[0107] Embodiments of the present disclosure will now be described, by way of example only, with reference to the following diagrams wherein:

[0108] FIG. 1 (a) illustrates a front-side view of the system (100), in accordance with an embodiment of the present disclosure;

[0109] FIG. 1 (b) illustrates a back-side view of the system (100), in accordance with an embodiment of the present disclosure;

[0110] FIG. 2 illustrates the system (100) with an external lateral flow strip (200) attached to the outlet (102), in accordance with an embodiment of the present disclosure;

[0111] FIG. 3 illustrates the capillary motion of the biofluid in the microfluidic channel (105), in accordance with an embodiment of the present disclosure;

[0112] FIG. 4 illustrates the microfluidic channel (105) of the system (100) with embedded gold- conjugated latex particles (108), in accordance with an embodiment of the present disclosure;

[0113] FIG. 5 (a) illustrates the system (100) with the lateral flow strip (200) that shows double lines (201) which indicates a positive result i.e. the presence of tuberculosis, in accordance with an embodiment of the present disclosure; FIG. 5 (b) illustrates the system (100) with the lateral flow strip (200) that shows a single control line (202) which indicates a negative result i.e. the absence of tuberculosis, in accordance with an embodiment of the present disclosure;

[0114] FIG. 5 (c) illustrates the system (100) with the lateral flow strip (200) that shows a single test line (203) which indicates the indeterminate result, in accordance with an embodiment of the present disclosure;

[0115] FIG. 6 (a) illustrates the system (100) without the lateral flow strip (200) that shows double lines

[0116] (109) which indicates a positive result i.e. the presence of tuberculosis, in accordance with an embodiment of the present disclosure; and

[0117] FIG. 6 (b) illustrates the system (100) without the lateral flow strip (200) that shows a single line

[0118] (110) which indicates a negative result i.e. the absence of tuberculosis, in accordance with an embodiment of the present disclosure.

[0119] In the accompanying drawing, an underlined number is employed to represent an item over which the underlined number is positioned or an item to which the underlined numberis adjacent. A nonunderlined number relates to an item identified by a line linking the non-underlined number to the item. When a number is non-underlined and accompanied by an associated arrow, the nonunderlined number is used to identify a general item at which the arrow is pointing.

[0120] DETAILED DESCRIPTION OF EMBODIMENTS

[0121] In the following detailed description, embodiments, of the present disclosure, will now be described with reference to the accompanying drawing, which forms a part hereof.

[0122] The following detailed description illustrates embodiments of the present disclosure and ways in which they can be implemented. Although some modes of carrying out the present disclosure have been disclosed, those skilled in the art would recognize that other embodiments for carrying out or practising the present disclosure are also possible.

[0123] Infectious diseases are widespread in the world. One of the most prominent and deadlier ones is Tuberculosis which is caused by Mycobacterium tuberculosis (MTB). According to the World Health Organization (WHO), the infectious disease tuberculosis kills about three million people every year. Globally, there are millions of people who suffer from tuberculosis. Millions die every year due to this disease. In developing countries, the disease is rampant. People living in poverty- stricken areas bear the greatest burden of the disease. Most of those affected are in the economically productive age group. There are several conventional tests for the detection of MTB, however, these conventional tests all have disadvantages. Further, there are other point-of-care tests (POCT) available for childhood tuberculosis such as the detection of Lipoarabinomannan (LAM) in urine using lateral flow assays, loop-mediated isothermal amplification (LAMP), PCR- based point-of-care tests and point-of-care ultrasound (POCUS) imaging for pulmonary tuberculosis, however, aforestated conventional tests and other point-of-care tests for childhood tuberculosis have are not up to the benchmark and have their own disadvantage such as high percentage of false positives, requires needle injection, poor patient compliance, takes longer time for detection, less specific, low sensitivity, expensive, complex in nature, requires attendance at health centres for several times before diagnosis, requires some technological expertise, and lots of infrastructure support and the like. In an aspect, the present disclosure provides a system (100) forthe detection of bacterial infection in the subject which overcomes the drawbacks of the conventional tests and provides a technical solution to the prior art. FIG. 1 (a) illustrates a front-side view of the system (100), in accordance with an embodiment of the present disclosure. FIG. 1 (b) illustrates a back-side view of the system (100), in accordance with an embodiment of the present disclosure.

[0124] The system (100) comprises a biofluid collector which consists of an outer surface (101) that is made up of biocompatible material for contacting a skin surface of a subject and a pressuresensitive polyurethane fdm (103), a microfluidic channel (105), at least one marker (106), optionally, a lateral flow test strip (200), an outlet for removing an excess biofluid (102).

[0125] FIG. 2 illustrates the system (100) with an external lateral flow strip (200) attached to the outlet (102), in accordance with an embodiment of the present disclosure.

[0126] The outer surface comprises an inlet (104) that receives the biofluid.

[0127] The lateral flow test strip (200) optionally is connected with the biofluid collector and microfluidic channel (105) through the outlet (102).

[0128] The microfluidic channel (105) is in fluid communication with the biofluid collector, the marker (105) and the lateral flow test strip (200).

[0129] The parts of the system (100) are arranged from one to other to form a single unit.

[0130] In accordance with an embodiment of the present disclosure, the bacterial infection is at least one selected from the group consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus. In an exemplary embodiment of the present disclosure, the bacterial infection is Tuberculosis.

[0131] In accordance with an embodiment of the present disclosure, the system (100) detects Tuberculosis, diagnose and monitors the infection in real-time.

[0132] In accordance with an embodiment of the present disclosure, the system (100) detects active and latent stages of Tuberculosis.

[0133] In accordance with an embodiment of the present disclosure, the subject is a human.

[0134] In accordance with an embodiment of the present disclosure, the system is a detachable device that is attached directly to the skin surface and the device has a length ranging from 5 to 10 cm and a width ranging from 0.5 cm to 2 cm.

[0135] In accordance with an embodiment of the present disclosure, the device is attached to the skin surface through a skin-safe adhesive selected from marker adhesive and paper adhesive. In accordance with an embodiment of the present disclosure, the biofluid is at least one selected from the group consisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid. In an exemplary embodiment of the present disclosure, the biofluid is sweat.

[0136] In accordance with an embodiment of the present disclosure, the biocompatible material of the outer surface (101) is a hydrophilic foam. In an exemplary embodiment of the present disclosure, the hydrophilic foam is highly permeable to the skin surface. In accordance with an embodiment of the present disclosure, the innermost surface of the biofluid collector is impermeable to the biofluid (sweat) which ensures the proper entrapment of the biofluid and retains the biofluid for further processing. Therefore, this avoids draining as well as drying out of the collected sweat.

[0137] In accordance with an embodiment of the present disclosure, the outer surface (101) covers an external surface area in the range of 20 mm2to 25 mm2.

[0138] In accordance with an embodiment of the present disclosure, the inlet (104) consists of pores that direct the flow of sweat and dispense it to the microfluidic channel (105) followed by optionally the lateral flow test strip (200).

[0139] In accordance with an embodiment of the present disclosure, the pores are characterized by having an average pore size of less than 0.1 mm.

[0140] FIG. 3 illustrates the capillary motion of the biofluid in the microfluidic channel (105), in accordance with an embodiment of the present disclosure;

[0141] In accordance with an embodiment of the present disclosure, the microfluidic channel (105) has an internal volume to trap the sweat ranging from 50 ul to 100 ul in a time period ranging from 5 to 10 minutes.

[0142] In accordance with an embodiment of the present disclosure, the system (100) further comprises a sensor (107) which is embedded in the microfluidic channel (105).

[0143] In accordance with an embodiment of the present disclosure, the sensor is a chemical sensor.

[0144] The chemical sensor detects the analyte of interest. The chemical sensor reacts with the biomarker in the sweat and produces a colourimetric / colour change if the biomarker is present. So the chemical sensor varies depending on the presence of the biomarker used and the intensity of the colour change is determined by the level of the biomarkers in the sweat.

[0145] The chemical sensor has specificity in the range of 80 to 90%, preferably 88% and sensitivity in the range of 90 to 95%, preferably 92%.

[0146] In accordance with an embodiment of the present disclosure, the biofluid in the microfluidic channel (105) has a unidirectional flow.

[0147] In accordance with an embodiment of the present disclosure, the marker (106) is human serum amyloid protein AA2.

[0148] FIG. 5 (a) illustrates the system (100) with the lateral flow strip (200) that shows double lines (201) which indicates the positive result i.e. the presence of tuberculosis, in accordance with an embodiment of the present disclosure. FIG. 5 (b) illustrates the system (100) with the lateral flow strip (200) that shows a single control line (202) which indicates the negative result i.e. the absence of tuberculosis, in accordance with an embodiment of the present disclosure.

[0149] FIG. 5 (c) illustrates the system (100) with the lateral flow strip (200) that shows a single test line (203) which indicates the indeterminate result, in accordance with an embodiment of the present disclosure;

[0150] In accordance with an embodiment of the present disclosure, the lateral flow test strip (200) detects the presence of the specific marker / protein associated with Tuberculosis.

[0151] In accordance with an embodiment of the present disclosure, the lateral flow test strip is 100% specific for detecting Mycobacterium tuberculosis .

[0152] FIG. 6 (a) illustrates the system (100) without the lateral flow strip (200) that shows double lines (109) which indicates the presence of tuberculosis, in accordance with an embodiment of the present disclosure. FIG. 6 (b) illustrates the system (100) without the lateral flow strip (200) that shows a single line (110) which indicates the absence of tuberculosis, in accordance with an embodiment of the present disclosure. In accordance with an embodiment of the present disclosure, the marker (106) is embedded into the microfluidic channel (105) for early detection of Tuberculosis directly from the sweat and / or saliva in the microfluidic channel.

[0153] In accordance with an embodiment of the present disclosure, the lateral flow test strip (200) comprises a specimen pad (203) having dispensed sweat and / or saliva, a conjugate pad (204) comprising at least one detector molecule, and a test line (199) comprising capture molecules.

[0154] In accordance with an embodiment of the present disclosure, each of the detector molecules comprises a first antigen-specific antibody conjugated to a detectable particle.

[0155] In accordance with an embodiment of the present disclosure, the detectable particle is gold nanoparticles.

[0156] In accordance with an embodiment of the present disclosure, the gold nanoparticles have a size in the range from 3 nm to 40 nm.

[0157] In accordance with an embodiment of the present disclosure, the detector molecules comprise one or more first antigen-specific antibodies conjugated to the detectable particle.

[0158] In accordance with an embodiment of the present disclosure, the first antigen-specific antibody is specific for Mycobacterium tuberculosis .

[0159] In accordance with an embodiment of the present disclosure, the capture molecules comprise a second antigen-specific antibody. In accordance with an embodiment of the present disclosure, the second antigen-specific antibody is specific for Mycobacterium tuberculosis .

[0160] In accordance with an embodiment of the present disclosure, the specimen pad contains a pretreated buffer and / or is added by a dropper.

[0161] In accordance with an embodiment of the present disclosure, the buffer is a SAA assay buffer.

[0162] In accordance with an embodiment of the present disclosure, the outlet (102) has a diameter in the range of 2 to 10 mm.

[0163] In accordance with an embodiment of the present disclosure, the system uses capillary motion to control the flow of sweat and / or saliva among its all components.

[0164] In accordance with an embodiment of the present disclosure, the system (100) provides test results in about 30 minutes or less. In accordance with an embodiment of the present disclosure, the system (100) provides test results in about 20 minutes or less. In accordance with an embodiment of the present disclosure, the system (100) provides test results in about 10 minutes.

[0165] In accordance with an embodiment of the present disclosure, the system (100) provides test results with an accuracy of about 80% or greater. In accordance with an embodiment of the present disclosure, the system (100) provides test results with an accuracy of about 85% or greater. In accordance with an embodiment of the present disclosure, the system (100) provides test results with an accuracy of about 88% or greater.

[0166] FIG. 4 illustrates the microfluidic channel of the system (100) with embedded latex particles, in accordance with an embodiment of the present disclosure. In accordance with an embodiment of the present disclosure, the microfluidic channel (105) is further embedded with latex particles (108) forthe detection of Tuberculosis.

[0167] In accordance with an embodiment of the present disclosure, the system (100) is safe, accurate and easy to handle without health workers.

[0168] In accordance with an embodiment of the present disclosure, the system (100) is non-invasive and allows rapid detection / screening of tuberculosis patients.

[0169] In accordance with an embodiment of the present disclosure, the system (100) uses specific antigen-antibody interaction.

[0170] In another aspect, the present disclosure provides a method for point-of-care detection of a bacterial infection.

[0171] The method comprises the following steps: a) obtaining a biofluid excreted from the skin of the subject by placing the system (100) of claim 1 on the skin surface of the subject for 10 min to collect the biofluid; b) dispensing the collected biofluid to a microfluidic channel (105) through an inlet (104); and c) allowing the dispensed biofluid in the microfluidic channel (105) for 15 to 30 minutes followed by detecting the presence and absence of the bacterial infection in the subject.

[0172] In accordance with an embodiment of the present disclosure, the bacterial infection is at least one selected from the group consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus. In an exemplary embodiment of the present disclosure, the bacterial infection is Tuberculosis.

[0173] In accordance with an embodiment of the present disclosure, the method detects Tuberculosis, diagnose and monitors the infection in real-time.

[0174] In accordance with an embodiment of the present disclosure, the method detects active and latent stages of Tuberculosis.

[0175] In accordance with an embodiment of the present disclosure, the subject is human.

[0176] In accordance with an embodiment of the present disclosure, the biofluid is at least one selected from the group consisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid. In an exemplary embodiment of the present disclosure, the biofluid is sweat. In another exemplary embodiment of the present disclosure, the biofluid is saliva.

[0177] In accordance with an embodiment of the present disclosure, the biofluid is dispensed from the inlet (104) to the microfluidic channel (105) and other components through a capillary motion.

[0178] In accordance with an embodiment of the present disclosure, the microfluidic channel (105) comprising a chemical sensor (107) wherein the chemical sensor (107) is exposed to the biofluid followed by measuring an output signal of the chemical sensor upon exposure to the biofluid.

[0179] In accordance with an embodiment of the present disclosure, the formation of double lines (109, 201) confirms the presence of Mycobacterium tuberculosis in the subject and wherein the formation of a single line (110, 202) confirms the absence of Mycobacterium tuberculosis in the subject.

[0180] In accordance with an embodiment of the present disclosure, the system (100) is placed on the chest, hands, legs, and inner arm skin of the subject.

[0181] In accordance with an embodiment of the present disclosure, a lateral flow strip (200) is optionally attached to the system (100) through the outlet (102).

[0182] In accordance with an embodiment of the present disclosure, a buffer solution is added to the specimen pad of the lateral flow strip (200) when the LFA is attached to the system (100) via outlet (102) to ensure migration of the biofluid along the path and preserve the marker of interest. The specimen pad is usually pretreated and in addition, there is a buffer solution provided with a dropper to add 2-3 drops, to the specimen pad after being fully wicked with the biofluid.

[0183] In accordance with an embodiment of the present disclosure, the method provides test results in about 30 minutes or less.

[0184] In accordance with an embodiment of the present disclosure, the method provides test results in about 20 minutes or less.

[0185] In accordance with an embodiment of the present disclosure, the method provides test results in about 10 minutes.

[0186] In accordance with an embodiment of the present disclosure, the method provides test results with an accuracy of about 80% or greater.

[0187] In accordance with an embodiment of the present disclosure, the method provides test results with an accuracy of about 85% or greater.

[0188] In accordance with an embodiment of the present disclosure, the method provides test results with an accuracy of about 88% or greater.

[0189] Modifications to embodiments of the present disclosure described in the foregoing are possible without departing from the scope of the present disclosure as defined by the accompanying claims. Expressions such as "including", “at least one”, "comprising", "incorporating", "have", "is" used to describe and claim the present disclosure are intended to be construed in a non-exclusive manner, namely allowing for items, components or elements not explicitly described also to be present. Reference to the singular is also to be construed to relate to the plural. The word "exemplary" is used herein to mean "serving as an example, instance or illustration". Any embodiment described as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments or to exclude the incorporation of features from other embodiments. The word "optionally" is used herein to mean "is provided in some embodiments and not provided in other embodiments". It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable combination or as suitable in any other described embodiment of the disclosure.

[0190] As used herein, the term "about", in the context of the time period of components, conditions, other measurement values, etc., means + / - 90% of the stated value, or + / - 95% of the stated value, or + / - 99% of the stated value, or < 30 minutes of the stated value, or < 20 minutes of the stated value, or < 10 minutes of the stated value. As used herein, the term “at least one” is equivalent to "one or more"." For example, a marker is at least one selected from the group consisting of colloidal gold nanoparticles and protein A-coated gold nanoparticles. That means the marker of the present disclosure can be one marker or more than one marker. Moreover, the use of the phrase “at least one” in claims is a worldwide accepted standard practice of claim drafting. This expression is definite and does not seek excessive protection.

[0191] Throughout this disclosure, certain embodiments may be disclosed in a range of formats. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosed ranges. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, a description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0192] The foregoing description of the embodiments has been provided for purposes of illustration and is not intended to limit the scope of the present disclosure. Individual components of a particular embodiment are generally not limited to that particular embodiment, but, are interchangeable. Such variations are not to be regarded as a departure from the present disclosure, and all such modifications are considered to be within the scope of the present disclosure.

[0193] The present disclosure is further described in light of the following experiments which are set forth for illustration purposes only and not to be construed as limiting the scope of the disclosure. The following experiments can be tested to scale up to an industrial / commercial scale and the results obtained can be extrapolated to the industrial scale.

[0194] EXPERIMENTS

[0195] Example 1: SAA assay Buffer preparation

[0196] 0.159 gm of sodium carbonate, 0.294 gm of sodium bicarbonate, 0.095 gm of sodium azide and 0.9 gm of sodium chloride were weighed independently and added in 250 ml of a beaker containing 80 ml of water to obtain a homogenous mixture. 1 ml of polysorbate 20 (Tween 20) was added to the so obtained mixture by adjusting the pH to 8 and followed by adding + / - 0.1 with 1 N HCL on a magnetic stirrer. The volume was made with distilled water to 100 ml followed by mixing to obtain the SAA assay buffer.

[0197] Example 2: Preparation of colloidal gold and Nitrocellulose membrane:

[0198] Colloidal gold was manufactured using the sodium citrate reduction method. Gold chloride Trihydrate (Sigma; Cat No G-4022), and Tri Sodium Citrate (Sigma; Cat No. S-4641) were used as raw materials.

[0199] For the capture molecules, colloidal gold was conjugated to Anti-SAS Ab (Cat No. ab207445), blocked with BSA (MP Biomedicals Cat No.: 160069) and re-suspended in proprietary colloidal gold conjugate buffer containing borate buffer, sucrose as principal components. Capture cone. (on NCM: cat.no. 207445): 0.426 mg / ml

[0200] For the control line, colloidal gold was conjugated to mouse IgG ( Arista Biologicals Cat No. AGMIG-0100), blocked with BSA (MP Biomedicals Cat No.: 160069) and re-suspended in proprietary colloidal gold conjugate buffer containing borate buffer, sucrose as principal components. Conjugate Ab (Cat No. 125414), the conjugate of OD 10 used for the control line goat, anti-mouse was used at 0.8 mg / ml and mouse IgG conjugate of OD 4 used

[0201] The anti-SAS antibody conjugated colloidal gold conjugate and mouse IgG colloidal gold were impregnated into a polyester conjugate release pad and dried for 6 hrs in a room maintained at RH of below 15%. The dried colloidal gold conjugate pads were stored in sealed aluminium foil pouches for further use.

[0202] Nitrocellulose Membrane NCPC -040 (MDI, India) was coated with Anti-SAS Ab (Cat No. ab 125414) as a test line and as a procedural control line. The antibody-coated membrane dried at 45°C for 1 hr in a controlled temperature oven. The coated membranes were stored in sealed aluminium foil pouches for further use.

[0203] The coated membrane pouches and conjugate pad pouches were opened in a room maintained at RH between 20- 30 %. The coated membrane was assembled with a conjugate pad strip, sample pad strip and absorbent pad. The assembled membranes were cut into strips. The strips were placed in a plastic housing assembly to obtain cassettes. The cassettes were sealed in an aluminium foil pouch along with a desiccant.

[0204] Example 3: Preparation of test line capture molecules:

[0205] Test line capture molecules were prepared by conjugating biotin to anti-antibodies (Millipore, Massachusetts, USA) according to the following protocol:

[0206] 1. 50mM EZ-link® Amine-PEG2 -Biotin (Thermo Scientific, Massachusetts, USA) was prepared in 0. 1 M MES buffer to form a Biotin-MES solution.

[0207] 2. 0.5 to 1ml of 0.1M MES buffer was added to 50uL o lOOuL (i.e., of the Biotin- MES solution.

[0208] 3. A sufficient volume of the antibody was then added to the solution to prepare a MES- Biotin-Antibody solution having a concentration of l.Omg / ml to 2.0mg / mL.

[0209] 4. 1 to 5uL of lOmM EDC (l-Ethyl-3-(3- dimethyl aminopropyl)carbodiimide) was added into the MES-Biotin-Antibody solution.

[0210] 5. The resultant solution was incubated for 2hrs with gentle shaking at room temperature.

[0211] 6. After incubation, the resultant solution was centrifuged at 14000g for lOmins and the biotin-antibody supernatant was collected.

[0212] 7. The biotin-antibody supernatant was then subjected to a desalting process: a. A desalting column was opened and inserted into a 15mL falcon tube before spinning it down once at 1000g for 2mins. b. The falcon tube was discarded. The desalting column was inserted into a new 15ml falcon tube and overlaid with the biotin-antibody supernatant obtained from step (6) and spun at 1000g for 2mins. c. The biotin-antibody supernatant was collected and the desalting column was discarded. d. The biotin-antibody supernatant collected was stored at -20°C until further use. Example 4: Preparation of detector molecules:

[0213] Detector molecules were prepared by conjugating protein A-coated colloidal gold (i.e., protein A-coated colloidal gold nanoparticles) to TB-specific markers according to the following protocol:

[0214] 1. 1.2ml of Colloidal gold solution comprising gold nanoparticles was added into an Eppendorf tube and spun at 4°C, 25000g for Ihr.

[0215] 2. The supernatant was discarded. The pellet was re-suspended with 1ml of Protein A IgG Binding Buffer (ThermoScientific).

[0216] 3. A sufficient volume of the selected biomarker antibody(ies) was added to the resultant mixture of step (2) to prepare a solution having a final concentration of 0.1 to 0.3mg / mL.

[0217] 4. The resultant solution of step (3) was incubated for 1 hour with gentle shaking on a thermomixer at 450rpm at 25 °C.

[0218] 5. After incubation, the solution was spun down at 4°C at 25000g for Ihr. The pellet was resuspended in 0.5 to ImL of Protein A IgG Binding Buffer (ThermoScientific) to the final solution comprising protein A coated gold nanoparticles conjugated to the antibody used.

[0219] 6. The final solution comprising protein A-coated gold nanoparticles conjugated to the antibody can be stored at 4°C until further use.

[0220] Example 5: Preparation of control line capture molecules:

[0221] Control line capture molecules were prepared by conjugating biotin to goat anti-mouse IgG antibodies (ThermoScientific) and ten streptavidin according to the following protocol:

[0222] 1. 50mM EZ-link® Amine-PEG2-Biotin (ThermoScientific) was prepared in 0.1 M MES buffer to form a Biotin-MES solution.

[0223] 2. 0.5 to ImL of 0 .IM MES buffer was added to 50uL to lOOuL of the Biotin-MES solution.

[0224] 3. A sufficient volume of goat anti -mouse IgG antibodies (ThermoScientific) was then added to the solution to prepare an MES-Biotin-Antibody solution having a concentration of 2.0mg / ml.

[0225] 4. 1 to 5uL of lOmM EDC was added into the MES-Biotin-Antibody solution.

[0226] 5. The resultant solution was incubated for 2hrs with gentle shaking at room temperature.

[0227] 6. After incubation, the resultant solution was centrifuged at 14000g for 1 Omins and the biotinantibody supernatant was collected.

[0228] 7. The biotin-antibody supernatant was then subjected to desalting: a. A desalting column was opened and inserted into a 15ml falcon tube before spinning it down once at 1000g for 2mins. b. The falcon tube was discarded. The desalting column was inserted into a new 15ml falcon tube and overlaid with biotin-antibody supernatant obtained from step (6) and spun at 1000g for 2mins. c. The biotin-antibody supernatant was collected and the desalting column was discarded. d. The biotin-antibody supernatant collected was stored at -20°C until further use. e. 100 to 500 pl of Img / ml Streptavidin (Sigma-Aldrich, St. Louis, Missouri, USA) was added to the biotin-antibody supernatant solution and mixed thoroughly. f. The final biotin-antibody solution was stored at -20°C until further use. Example 6: Evaluation of clinical efficacy in diseased patients and healthy patients

[0229] An experiment was conducted on diseased patients who were diagnosed with confirmed TB and healthy subjects to ascertain the specificity, sensitivity and efficacy of the system. The age range of the cohort was 19-49 years and male to female ratio of 1.5: 1.

[0230] Aims and objectives: To ascertain the yield, prevalence and number needed to screen (NNS) to detect a case of TB through household contact investigation within the intervention areas.

[0231] Method:

[0232] A prospective case-controlled study was conducted with 40 individuals in which 25 TB patients confirmed with sputum smear and GeneXpert (a molecular detection of TB that detects DNA in TB bacteria and using sputum sample) and 15 healthy controls. Institutional Ethical approval was obtained and participants also gave informed written consent. The strips (i.e. system- a detachable device in accordance with the present disclosure) were used to collect sweat and or saliva (i.e. a biofluid) for 10 minutes from each participant and processed for 25 minutes as per the protocol. Results were read as negative (110, 202- figures 5b and 6b), positive (109, 201- figures 5a and 6a) and indeterminate (20- figure 5c).

[0233] Results:

[0234] The test was positive in 22 patients (88%) out of 25 TB-confirmed patients with PTB (pulmonary tuberculosis) and negative in 2 (8%) patients with PTB. There was an indeterminate result in one patient. Among 15 apparently healthy individuals, who were negative for PTB and have no symptoms of TB, the test was negative in 12 (80%) individuals, positive in 2 (13%) and indeterminate in 1 person (7%), thus p-value <0.05.

[0235] The test was clinically effective for the detection of PTB without using any sputum sample or any other invasive procedure, specific i.e. accurate (88%), easy to use and rapid (less than 10 minutes) and sensitive. All the participants and health workers preferred the test over the conventional test methods that used sputum and blood or any other sample to detect TB.

[0236] The preliminary clinical trial of the system (a detachable device-strips) for the diagnosis of pulmonary tuberculosis was effective in patients with a greater rate of acceptance / compliance and performance.

[0237] A similar clinical experiment was also conducted with the same group of patients using the lateral flow test strip (200) connected with the biofluid collector and the microfluidic channel through the outlet (102).

[0238] It was found that the same results were obtained as that of above.

Claims

CLAIMS1. A system (100) for the detection of a bacterial infection in a subject, the system (100) comprising: a) a biofluid collector comprises: i. an outer surface (101) consisting of a biocompatible material for contacting a skin surface of the subject; and ii. a pressure-sensitive polyurethane film (103), b) a microfluidic channel (105); c) at least one marker (106); d) optionally, a lateral flow test strip (200); e) an outlet for removing an excess biofluid (102), wherein the outer surface comprises an inlet (104) for receiving the biofluid, wherein optionally, the lateral flow test strip (200) is connected with the biofluid collector and the microfluidic channel through the outlet (102), wherein the microfluidic channel (105) is in fluid communication with the biofluid collector, said marker (106) and said lateral flow test strip (200), and wherein said parts (a) to (e) of the system are arranged from one to other to form a single unit.

2. The system (100) of claim 1, wherein the bacterial infection is at least one selected from the group consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus.

3. The system (100) of claim 2, wherein the bacterial infection is Tuberculosis.

4. The system (100) of claim 3, wherein the system (100) detects Tuberculosis, diagnose and monitors the infection in real-time.

5. The system (100) of claim 4, wherein the system (100) detects active and latent stages of Tuberculosis.

6. The system (100) of claim 1, wherein the subject is a human.

7. The system (100) of claim 1, wherein the system (100) is a detachable device that is attached directly to the skin and wherein the device has a length ranging from 5 to 10 cm and a width ranging from 0.5 cm to 2 cm.

8. The system (100) of claims 1 and 7, wherein the device is attached to the skin through a skin-safe adhesive selected from marker adhesive and paper adhesive.

9. The system (100) of claim 1, wherein the biofluid is at least one selected from the groupconsisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid.

10. The system (100) of claim 9, wherein the biofluid is selected from sweat and / or saliva.

11. The system ( 100) of claim 1 , wherein the biocompatible material of the outer surface (101) is a hydrophilic foam.

12. The system (100) of claim 11, wherein the hydrophilic foam is highly permeable to the skin surface.

13. The system (100) of claim 1, wherein the outer surface (101) covers an external surface area in the range of 20 mm2to 25 mm2.

14. The system (100) of claim 1, wherein the inlet (104) consists of pores that direct the flow of the biofluid and dispense it to the microfluidic channel (105) followed by optionally the lateral flow test strip (200).

15. The system (100) of claim 14, wherein the pores are characterized by having an average pore size of less than 0.1 mm.

16. The system (100) of claim 1, wherein the microfluidic channel (105) has an internal volume to trap the sweat ranging from 50 ul to 100 ul in atime period ranging from 5 to 10 minutes.

17. The system (100) of claim 1, wherein the system further comprises a sensor (107) which is embedded in the microfluidic channel (105).

18. The system (100) of claim 17, wherein the sensor is a chemical sensor and wherein the sensor has a sensitivity in the range of 90 to 95% and specificity in the range of 80 to 90%.

19. The system (100) of claim 1, wherein the biofluid in the microfluidic channel (105) has a unidirectional flow.

20. The system (100) of claim 1, wherein the marker (106) is human serum amyloid protein AA2.

21. The system (100) of claim 20, wherein the marker (106) is embedded into the microfluidic channel (105).

22. The system (100) of claim 1, wherein the lateral flow test strip (200) comprises a specimen pad (203) having dispensed sweat and / or saliva, a conjugate pad (204) comprising at least one detector molecule, and a test line (199) comprising capture molecules.

23. The system (100) of claim 22, wherein the lateral flow test strip (200) detects the presenceof the specific marker / protein associated with Mycobacterium tuberculosis .

24. The system (100) of claims 22 or 23, wherein the lateral flow test strip is 100% specific for detecting Mycobacterium tuberculosis .

25. The system (100) of claim 22, wherein each of the detector molecules comprises a first antigen-specific antibody conjugated to a detectable particle.

26. The system (100) of claim 25, wherein the detectable particle is gold nanoparticles.

27. The system (100) of claim 26, wherein the gold nanoparticles have an average size in the range from 3 nm to 40 nm.

28. The system (100) of claims 22 to 27, wherein each of the detector molecules comprises one or more first antigen-specific antibodies conjugated to the detectable particle.

29. The system (100) of claim 28, wherein the first antigen-specific antibody is specific for Mycobacterium tuberculosis .

30. The system (100) of claim 22, wherein the capture molecules comprise a second antigenspecific antibody.

31. The system (100) of claim 30, wherein the second antigen-specific antibody is specific for Mycobacterium tuberculosis .

32. The system (100) of claim 22, wherein the specimen pad contains a pretreated buffer and / or is added by a dropper.

33. The system (100) of claim 32, wherein the buffer is SAA assay buffer.

34. The system (100) of claim 1, wherein the outlet (102) has a diameter in the range of 2 to 10 mm.

35. The system (100) of claim 1, wherein the system uses capillary motion to control the flow of sweat among its all components.

36. The system (100) of claim 1, wherein the system provides test results in 30 minutes or less.

37. The system (100) of claim 36, wherein the system provides test results in 20 minutes or less.

38. The system (100) of claim 36, wherein the system provides test results in 10 minutes.

39. The system (100) of claim 1, wherein the system provides test results with an accuracy of80% or greater.

40. The system (100) of claim 39, wherein the system provides test results with an accuracy of 85% or greater.

41. The system (100) of claim 39, wherein the system provides test results with an accuracy of 88% or greater.

42. The system (100) of claim 1, wherein the microfluidic channel (105) is further embedded with gold-conjugated latex particles (108) forthe detection of Mycobacterium tuberculosis .

43. The system (100) of claim 1, further comprising (a) a processing unit, which receives an output signal of the sensor and detects said signal qualitatively and quantitatively and (b) a transmitter, which receives an output signal of the sensor and transmits said signal to a remote server and / or to a portable electronic device.

44. The system (100) of claim 43, wherein the remote server and the portable electronic device comprise an algorithm, which generates qualitative and quantitative data for further diagnosing and / or monitoring.

45. The system (100) of any one of claims 43 to 44, comprising an indicator, wherein the processing unit and the transmitter are configured to transmit a positive indication signal or a negative indication signal to the indicator, thereby displaying the outcome.

46. The system (100) of any one of claims 43 to 45, wherein the processing unit, the remote server and the portable electronic device further comprises an algorithm configured to differentiate between the response of the sensor to the biofluid excreted from the skin and the response of the sensor to a stimulus selected from the group consisting of temperature, humidity, lateral strain, and combinations thereof.

47. The system (100) of any one of claims 43 to 46, wherein the system generates real-time quantitative and qualitative data for early detection, diagnosis and / or monitoring of Mycobacterium tuberculosis in the subject.

48. A method for point-of-care (POC) detection of a bacterial infection in a subject, wherein the method comprising the steps of: a) obtaining a biofluid excreted from the skin of the subject by placing the system (100) of claim 1 on the skin surface of the subject for 10 min to collect the biofluid; b) dispensing the collected biofluid to a microfluidic channel (105) through an inlet (104); and c) allowing the dispensed biofluid in the microfluidic channel (105) for 15 to 30 minutes followed by detecting the presence and absence of the bacterial infection in the subject.

49. The method of claim 48, wherein the bacterial infection is at least one selected from thegroup consisting of Tuberculosis, Anthrax, Tetanus, Pneumonia, Cholera, Diphtheria, Typhoid and Typhus.

50. The method of claim 49, wherein the bacterial infection is Tuberculosis.

51. The method of claim 48, wherein the method detects Tuberculosis, diagnose and monitors the infection in real-time.

52. The method of claim 48, wherein the method detects active and latent stages of Tuberculosis.

53. The method of claim 48, wherein the subject is human.

54. The method of claim 48, wherein the biofluid is at least one selected from the group consisting of sweat, saliva, urine, seminal fluids, vaginal fluids, mucus, phlegm, blood, plasma, tears, gastrointestinal fluid, cerebrospinal fluid and synovial fluid.

55. The method of claim 54, wherein the biofluid is selected from sweat and / or saliva.

56. The method of claim 48, wherein the biofluid is dispensed from the inlet (104) to the microfluidic channel (105) and other components through a capillary motion.

57. The method of claim 48, wherein the microfluidic channel (105) comprising a chemical sensor (107) wherein the chemical sensor (107) is exposed to the bio fluid followed by measuring an output signal of the chemical sensor upon exposure to the biofluid.

58. The method of claims 48 to 53, wherein the formation of double lines (109, 201) confirms the presence of Mycobacterium tuberculosis in the subject, wherein the formation of a single control line (110, 202) confirms the absence of Mycobacterium tuberculosis in the subject and wherein the formation of a single test line (203) confirms an indeterminate result.

59. The method of claim 48, wherein the system (100) is placed on the chest, hands, legs, and inner arm skin of the subject.

60. The method of claim 48, wherein a lateral flow strip (200) is optionally attached to the system (100) through the outlet (102) for detection.

61. The method of claim 48, wherein the method provides test results in 30 minutes or less.

62. The method of claim 61, wherein the method provides test results in 20 minutes or less.

63. The method of claim 61, wherein the method provides test results in 10 minutes.

64. The method of claim 48, wherein the method provides test results with an accuracy of 80% or greater.

65. The method of claim 64, wherein the method provides test results with an accuracy of 85% or greater.

66. The method of claim 64, wherein the method provides test results with an accuracy of 88% or greater.

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