WO2022113112A1

WO2022113112A1 - Device for rna extraction for sars-cov-2 detection

Info

Publication number: WO2022113112A1
Application number: PCT/IN2021/051105
Authority: WO
Inventors: Ravikrishnan ELANGOVAN; Dinesh Kalyanasundaram; Manoj Balakrishna MENON; Vivekanandan PERUMAL; Ada ZWETLANA; Kanishka GROVER; Vineet PENUMARTHY; Vinod Parmar; Allanki Sai SWAROOP; Ujjwal DHAIYA; Tushar JEET; Prabhu Balasubramanian
Original assignee: Reliance Industries Limited
Priority date: 2020-11-27
Filing date: 2021-11-26
Publication date: 2022-06-02

Abstract

The present subject matter relates to a device (100) for extracting Ribonucleic Acid (RNA) from a cell sample. The device (100) includes a first storage unit L (118) having a pre-stored lysis reagent (R1) and an extraction unit (122) to receive the lysed cell sample from the first storage unit L (118) and to bind RNA out of the lysed cell sample to a filter (124) thereof. The extraction unit (122) is to further receive a wash reagent (R2) via to wash out inhibitors from the RNA and furthermore receives an elusion reagent (R3) to elude the washed RNA from the filter (124). The device (100) further includes an analysis unit (140) to receive the eluded RNA for nucleotide amplification.

Description

DEVICE FOR RNA EXTRACTION FOR SARS-CoV-2 DETECTION

FIELD OF INVENTION

[0001 ] The present subject matter relates, in general, to devices for invitro processing of nucleic acids, and, particularly, to devices for extracting Ribonucleic Acid (RNA) and from a cell sample for Severe Acute Respiratory Syndrome Coronavims-2 (SARS-CoV-2) virus detection.

BACKGROUND

[0002] Coronavirus is a type of Severe Acute Respiratory Syndrome

Coronavims-2of common RNA virus that infects human. Out of all the viruses belonging to the category of the coronavirus, “(SARS-CoV-2)” virus is responsible for causing the respiratory disease Coronavirus Disease 2019 (COVID-19). The SARS-CoV-2 virus interacts with Angiotensin-converting enzyme-2 (ACE-2) receptors on surfaces of cells of a respiratory system of the human to cause viral infection that leads to respiratory diseases or lung diseases. For identifying the viral infection of SARS-CoV-2 in a human body, antibodies generated in the human body in response to the viral infection are detected or the presence of a viral RNA in a testing sample is detected.

BRIEF DESCRIPTION OF DRAWINGS

[0003] The detailed descriptions are depicted with reference to the accompanying figures. In the figures, the left-most digit(s) of a reference number identifies the figure in which the reference number first appears. The same numbers are used throughout the figures to reference like features and components. Some implementations of the system(s), in accordance with the present subject matter, are described by way of examples, and with reference to the accompanying figures, in which:

[0004] FIG.l illustrates a schematic top view of a device for extracting

Ribonucleic Acid (RNA) followed by isothermal amplification analysis from a cell sample, according to an example implementation of the present subject matter; [0005] FIG.2A illustrates an isometric view of a device for extracting

Ribonucleic Acid (RNA) followed by isothermal amplification analysis from a cell sample, according to an example implementation of the present subject matter;

[0006] FIG.2B illustrates an isometric view of a device for extracting Ribonucleic Acid (RNA) followed by isothermal amplification analysis from a cell sample, according to an example implementation of the present subject matter;

[0007] FIG. 3 illustrates an isometric view, a top view, and a front view of an extraction unit of the device of FIG. 1, according to an example implementation of the present subject matter; [0008] FIG. 4 illustrates an isometric view, a top view, and a front view of an extraction unit connected with the analysis unit of the device of FIG. 1, according to an example implementation of the present subject matter; and

[0009] FIG. 5 illustrates an isometric view, a top view and a front view of a waste chamber of the device of FIG. 1, according to an example implementation of the present subject matter.

DETAILED DESCRIPTION

[0010] Generally, infection by a Severe Acute Respiratory Syndrome

Coronavims-2 (SARS-CoV-2) virus in a human body is determined by performing either of a serological test or a molecular test.

[0011 ] The serological test is performed by collecting a blood sample of a patient and then identifying the availability of “spike protein” in the blood sample. Since, the “spike protein” is highly immunogenic, an immune response in a human body is provoked to generate the antibodies in response to and counteracting an antigen related to the SARS-CoV-2 virus. However, the serological test fails to offer detection of the infection at an early stage of the infection, i.e., 1-2 days following the infection, since the production of the antibodies specific to the SARS- CoV-2 vims begins only 9 to 11 days after the patient begins to show symptoms. Therefore, the applicability of the serological test in detecting the infection due to the SARS-CoV-2 virus is limited as the viral infection cannot be detected at the earlier stage and can only be detected at an acute phase of the infection. Also, collecting, transporting and processing the blood sample for conducting the serological test requires a lot of effort and has a huge turnaround time for providing the result.

[0012] The molecular test, such as a reverse-transcriptase polymerase chain reaction (RT-PCR) is performed by collecting a sample of mucus from the nose or the throat of the patient. Rather than testing for antibodies, a genetic material or a genome of a virus causing the infection is detected from the collected sample. The primers, indicative of the known sequences of the genome, are added to the sample, where RNA of the virus is converted to DNA of the virus and genomic replication is emulated by addition of the relevant enzymes, cofactors, and substrates. The genomic replication allows growth of the DNA at the primer sequences. This process leads to creation of enough copies of the DNA, to which a fluorescent dye is added. The fluorescent dye enables the detection of the DNA structure of the genome of the virus. The RT-PCR involves a high instrumentation cost, requires significant training of the instruments used, has a high turnaround time, and has a high cost-per-test. Therefore, the RT-PCR based molecular testing is ineffective in conducting mass testing efforts for low-and middle-income populations.

[0013] A reverse-transcriptase loop-mediated isothermal amplification

(RT-LAMP) test is another molecular test that has seen widespread scientific application for detection of infection by a SARS-CoV-2 virus. However, RT- LAMP test requires highly trained personnel to carry out a multi-step testing process involving highly technical RNA extraction, sample purification, and RT- LAMP assay procedures. This again limits the reach of this form of testing to centralized laboratories in urban and semi-urban places.

[0014] The above-described conventional techniques may detect the SARS-

CoV-2 virus in the human body, but the turnaround time and cost required to execute the detection of the SARS-CoV-2 virus are high. Also, collection and transportation of the sample to testing facilities require a lot of effort. The rate of testing or detection of the SARS-CoV-2is required to be very high and inexpensive along with easy sample processing, sample purification, and sample transportation so that the further spread of the SARS-CoV-2 vims can be curtailed as quickly as possible.

[0015] The present subject matter describes devices for extracting

Ribonucleic Acid (RNA) from a cell sample, which facilitates transportation and processing of the cell sample, for e.g., a nasopharyngeal sample or a nasal sample for detection the SARS-CoV-2 virus in an economical manner and with an automated cell sample processing.

[0016] In an example implementation of the present subject matter, a device is provided with a first storage unit (L), a second storage unit (W), a third storage (E), an extraction unit, a waste chamber and an analysis unit. The first storage unit (L) includes a lysis reagent pre-stored therein. The lysis reagent enables the disintegration of RNA from the cell sample collected from a patient. In the first storage unit (L), the pre-stored lysis reagent is mixed with the cell sample for lysis thereof when the storage unit (L) receives the cell sample. The cell sample is a biological sample. In an example, the cell sample may be one of a cheek swab, a nasal swab, saliva and a throat gargle. The extraction unit further includes a first inlet, a second inlet, a third inlet, a first outlet, a second outlet, and a filter. The first inlet connects the first storage unit (L) and the extraction unit. The extraction unit receives the lysed cell sample from the storage unit via the first inlet. The porous filter of the extraction unit is a porous filter that binds specifically to RNA. In an example, the filter is a membrane that binds specifically to RNA. After receiving the lysed cell sample from the first storage unit (L), the lysed cell sample is then collected onto the filter, which is having a specific property of binding only to RNA. Due to such a specific property, the filter binds RNA out of the lysed cell sample, the residual reagent and the cell sample are transferred to the waster chamber of the device. The extraction unit further receives a wash reagent via the second inlet thereof. The wash reagent is pre-stored within the device in the second storage unit (W), which is connected to the extraction unit at the second inlet thereof. The wash reagent washes out the inhibitors from the RNA, which can inhibit nucleotide amplification of the RNA during the optical study of the RNA.. When the RNA is treated with the wash reagent and the inhibitors are washed out from the RNA, the extraction unit receives an elusion reagent via the third inlet thereof. The elusion reagent is pre- stored within the device in the third storage unit (E), which is connected to the extraction unit at the third inlet thereof. The elusion reagent eludes or dislodges the washed RNA from the filter so that nucleotide amplification of the RNA can be carried out.

[0017] Further, the analysis unit connected to the first outlet of the extraction unit receives the eluded RNA from the extraction unit. The analysis unit further includes a plurality of dried membranes for nucleotide amplification, to each of which the eluded RNA is diverted. Each dried membrane includes an amplification reagent to act upon the received RNA for nucleotide amplification so that the vims type present in the cell sample can be detected.

[0018] The device of the present subject matter functions as a lab-on-chip system, which integrate the full gamut of sample processing steps into a single microfluidic chip. The device reduces the test complexity, scope of error, and the volume of reagents required for sample processing. By this the turnaround time related to efficient fluid transport, sample processing, and RNA extraction is also reduced along with reduction in costs-per-test. Using the device of the present subject matter, the cell sample processing steps are automated.

[0019] Since all the reagents are pre-stored in the device, no reagent addition step is to be conducted manually. Also, the device of the present subject matter is a contained unit, once the cell sample is added no liquid material moves in or out for the vims detection.

[0020] These and other advantages of the present subject matter would be described in a greater detail in conjunction with FIGS. 1 to 5 in the following description. The manner in which the sample collection device is implemented and used shall be explained in detail with respect to FIGS. 1 to 5. It should be noted that the description merely illustrates the principles of the present subject matter. It will thus be appreciated that those skilled in the art will be able to devise various arrangements that, although not explicitly described herein, embody the principles of the present subject matter and are included within its scope. Furthermore, all examples recited herein are intended only to aid the reader in understanding the principles of the present subject matter. Moreover, all statements herein reciting principles, aspects and implementations of the present subject matter, as well as specific examples thereof, are intended to encompass equivalents thereof.

[0021 ] FIG. 1 illustrates a schematic top view of a device 100 for extracting

Ribonucleic Acid (RNA) from a cell sample, according to an example implementation of the present subject matter. The device 100 is injection moldable. The cell sample is a biological sample. In an example, the cell sample is one of a cheek swab, nasal swab, saliva, and throat gargle. The device 100 has a quadrilateral structure, in which a first side SI of the device 100 is provided with a first air inlet 102, a second air inlet 104, and a third air inlet 106. The first side SI of the device 100 includes a first air outlet 108 and a second air outlet 110. The first air inlet 102, the second air inlet 104, the third air inlet 106 are formed to receive a positive pressure. In an example, the positive pressure can be applied at the first air inlet 102, the second air inlet 104, the third air inlet 106 by means of an external device (not shown), for example, an air injection device. The first air outlet 108 and the second air outlet 110 are formed to receive a negative pressure. In an example, the negative pressure can be applied at the first air outlet 108 and the second air outlet 110 by means of another external device (not shown), for example, an air suction device. The first air inlet 102, the second air inlet 104, the third air inlet 106, the first air outlet 108, and the second air outlet 110, in each case, are air-tightly sealed in a non-operational state by a glue for sealing thereof in an air-tight manner. The glue is a medium that is used for sealing any opening. In an example, the glue is thermoplastic polyurethane glue. In an example, the glue is pierceable and is formed to keep the air-tight sealing intact after the piercing. The device 100 further includes a passage 112 having a first opening 114. The first opening 114 is formed at a proximal end of the passage 112 and is capable of receiving a cell sample collection device (not shown). The passage 112 has a second opening 116 at its distal end and concludes towards a first storage unit (L) 118 of the device 100. A longitudinal dimension of the passage 112 is parallel to a second side S2 of the device 100. The passage 112 is perpendicular to the first side SI of the device 100. A first air inlet 102 is connected to the passage 112 and is formed adjacent to the first opening 114. The third air inlet 106 is to enable inflow of air inside the first storage unit (L) 118 when the positive pressure is applied by an external air injection unit (not shown). The first storage unit (L) 118 is formed such that two sides of the first storage unit (L) 118 are adjacent to the second side S2 and a third side S3 of the device 100, such that the device 100 and the first storage unit (L) 118 have one common corner. The third side S3 is opposite to the first side SI. A lysis reagent R1 is pre-stored inside the first storage unit (L) 118. The lysis reagent R1 is shown with squares in FIG. 1 within the first storage unit (L) 118 of the device 100. The lysis reagent R1 is to carry out the lysis of the cell sample received in the first storage unit (L) 118. In the lysis, the outer membrane of the cell sample is disintegrated. In an example, the lysis reagent R1 is a lysis buffer solution. In an example, the lysis reagent R1 is 3 M Guanidium Hydrocholoride/Guanidium Thiocynate/Sodium Iodide and 40% Isoproponol or Ethanol, 100 mM Sodium acetate, 1-5% TritonXIOO or Tween20 or SDS and 5-20 mM DTT or beta meracpato ethanol or EDTA. In an example, any solution capable of lysis of a biological sample can be used as the lysis buffer solution.

[0022] Further, a capillary tube 120 connects the first storage unit (L) 118 to an extraction unit 122. The capillary tube 120 is configured to transmit a lysed cell sample towards the extraction unit 122 and is formed such that an automatic/accidental transmission of the lysed cell sample from the first storage unit (L) 118 to the extraction unit 122 is prevented. In an example, a reservoir (not shown) is abutted to the extraction unit 122. The extraction unit 122 includes a filter 124 having a porosity in a range from 500 nanometre (nm) to 5000 nm and a specific chemical property to allow binding or holding of RNA out of the lysed cell sample. In an example, the filter 124 is a semi-permeable membrane made of a silicone coated glass fibre. The extraction unit 122 is to extract RNA from the lysed cell sample received from the first storage unit (L) 118. The extraction unit 122 includes a first inlet 126, a second inlet 128, and a third inlet 130. The extraction unit 122 is in fluidic communication with a separating unit 132 having a first outlet 134 and a second outlet 136. In an example the separating unit 132 is a knob that functions as a three-way valve to control the direction of the cell sample in a desired direction. The separating unit 132 has a first tubular extension 138 at the first outlet 134 and is in fluidic communication with an analysis unit 140 of the device 100. The separating unit 132 has a second tubular extension 142 at the second outlet 136 and is in fluidic communication with a waste chamber 144 of the device 100. The waste chamber 144 is connected to the second outlet 136 of the extraction unit 122. The analysis unit 140 is connected to the first outlet 134 of the extraction unit 122. The analysis unit 140 includes a plurality of dried membranes 146 for nucleotide amplification. In an example, the analysis unit 140 includes three dried membranes. In an example, the analysis unit 140 includes one dried membrane. The analysis unit 140 is to receive the eluded RNA from the extraction unit 122 and to divert the eluded RNA to each of the plurality of dried membranes 146. Each of the dried membrane 146 comprises an amplification reagent so that the RNA when treated with the amplification reagent can undergo nucleotide amplification. The analysis unit may include a divertor (not shown) to equally divert the RNA towards the plurality of dried membranes. The analysis unit 140 is designed to have a single input from the second tubular extension 142 and to have multiple outputs. In an example, each of the dried membrane 146 is a Loop-Mediated Isothermal Amplification (LAMP) membrane. In an example, the amplification reagent may be a LAMP reagent. The LAMP membrane has a length and a width of about 2 millimetres and height of about 0.70 millimetres. The LAMP membrane has a thickness of about 400 micrometres.

[0023] In addition, the device 100 has a plurality of LAMP membrane chambers 148, each with a width of about 2 millimetres to accommodate each of the dried membrane 146.

[0024] Lurther in LIG.l, the waste chamber 144 is further connected to the first air outlet 108. The analysis unit 140 is connected with the second air outlet 110. The waste chamber 144 is to collect the lysed cell sample and residual reagents after the RNA binds to the filter 124.

[0025] The device 100 further includes a second storage unit (W) 150 connected at one end to the first air inlet 102 of the device and at another end to the second inlet 128 of the extraction unit 122. In an example, the second storage unit (W) 150 is connected to the second inlet 128 of the extraction unit 122 by means of a capillary tube 152. In an example, the second storage unit (W) 150 is capable of storing multiple buffer solutions. In an example, the second storage unit (W) 150 is pre-stored with a wash reagent R2 shown by triangles in FIG.l within the second storage unit (W)150. The wash reagent R2 is to wash out the inhibitors from the RNA. In an example, the wash reagent R2 is 80% Ethanol. In an example, the wash reagent R2 is 100% Isoproponol. The second storage unit (W) 150 is to supply the wash buffer R1 to the extraction unit 122.

[0026] Although, the second storage unit (W) 150 is depicted to have a serpentine shape, the shape of the second storage unit (W) 150 may vary. Further, the second storage unit (W) 150 is depicted as a single channel moulded in layers having convex curves, in an example embodiment of the present subject matter the number of channels may vary, where each channel may be connected to a next channel via a connection mechanism having air or oil plug gaps. The longitudinal dimensions of the second storage unit (W) 150 are parallel to the fourth side S4 of the device 100. The fourth side S4 is parallel to the second side S2.

[0027] In an example, the first air inlet 102, the second air inlet 104, the third air inlet 106, the first air outlet 108, and the second air outlet 110 are provided with filters (not shown) having porosity in a range of 10 micrometres to 50 micrometres. In an example implementation, the filters have a porosity of 20 micrometres.

[0028] The device 100 further includes a third storage unit (E) 154 connected at one end to the second air inlet 104 of the device 100 and at another end to the third inlet 130 of the extraction unit 122. In an example, the third storage unit (E) 154 is connected to the third inlet 130 of the extraction unit 122 by means of a capillary tube 156. In an example, the third storage unit (E) 154 is capable of storing multiple buffer solutions. In an example, the third storage unit (E) 154 is pre-stored with an elusion reagent R3 shown by circles in FIG.l within the third storage unit (E) 154. The third storage unit (E) 154 is to supply the elusion buffer to the extraction unit 122. The elusion reagent R3 is to elude the RNA, which is bind to the filter 124. In an example, the elusion reagent R3 is nuclease free water. In an example, the third storage unit (E) 154 is formed to store multiple buffer solutions, such that multiple solutions are prevented to get mix with each other.

[0029] Although, the third storage unit (E) 154 is depicted to have a serpentine shape, the shape of the third storage unit (E) 154 may vary. Further, the third storage unit is depicted as a single channel moulded in layers having convex curves, in an example embodiment of the present subject matter the number of channels may vary, where each channel may be connected to a next channel via a connection mechanism having air or oil plug gaps. The longitudinal dimensions of the third storage unit (E) 154 are parallel to the fourth side S4 of the device 100.

[0030] Further, the lysis reagent Rl, the wash reagent R2, and the elusion reagent R3 pre-stored in the device 100 are retained within the respective storage units (L, W, and E) by holding the air into the respective storage unit and the respective capillary tube at the other end. Thus, the spilling of the lysis reagent, the wash reagent, and the elusion reagent within the device 100 is prevented.

[0031] In operation, the cell sample collection device is inserted in the device 100 through the first opening 114 of the passage 112. The cell sample collection device moves towards the first storage unit (L) 118 via the passage 112. In an inserted state of the cell sample collection device in the device 100, a portion of the cell sample collection device, containing the cell sample, is placed inside the first storage unit (L) 118. In an example implementation another portion of the cell sample collection device has a square extrusion. The square extrusion is to connect to the first opening 114 of the device 100 in a form- fitting manner and lock the cell sample collection device in the device 100. The form-fitting connection creates an air-tight seal. Further, the lysis buffer Rl of the first storage unit (L) 118 disintegrates the RNA from the cell sample. Further, a positive pressure is applied at the third air inlet 106 and a negative pressure is applied at the first air outlet 108. In an example, the first air outlet 108 is opened to be exerted upon with a normal atmospheric pressure. Application of the positive pressure and the negative pressure flushes the lysed cell sample containing disintegrated RNA towards the extraction unit 122 via the capillary tube 120. The cell sample containing disintegrated RNA is further washed by the wash buffer R2 on the application of a positive pressure at the first air inlet 102 and a negative pressure at the first air outlet 108. Due to the positive pressure and the negative pressure, the second storage unit (W) 150 supplies the wash buffer R2 to the extraction unit 122 to wash out inhibitors from the RNA. In an example, the first air outlet 108 is opened to be exerted upon with a normal atmospheric pressure. The impurities, such as inhibitors are washed by the wash buffer R2 and pass through the filter 124 of the extraction unit 122 to get collected in the waste chamber 144. The RNA is bound on the filter 124 after impurities are washed off. After the RNA bound on the filter 124, the application of the positive pressure and the negative is stopped.

[0032] Further during the operation, the RNA bound at the filter 124 is released or eluded by the elution reagent R3 stored in the third storage unit (E) 154 on the application of a positive pressure at the second air inlet 104 and a negative pressure at the second air outlet 110. Due to the positive pressure and the negative pressure, the eluded RNA from the filter 124 is guided towards the analysis unit 140. The analysis unit 140 diverts the eluded RNA towards the plurality of dried membranes 146 for the nucleotide amplification when treated with the amplification reagent of each dried membrane. The pressure in the device 100 is created in the form of pulses or a continuous pressure may also be exerted. A pre calculated pressure is exerted to make the cell sample move to a pre-calculated distance within the device 100.

[0033] The device 100 is further synchronized with an external optical detection system. The device 100 is placed within the external optical detection system, such that the plurality of dried membranes 146 containing the released RNA are able to be acted upon by an excitation unit, a heating unit and a detection unit of the external optical detection system. Each of the plurality of dried membranes 146 is heated from an upper side as well as a lower side of the device 100. In an example, the plurality of dried membranes 146 are placed inside a transparent structure that does not interfere with heating process, excitation process and detection process by the external optical detection system. In an example, the transparent structure is formed from a material selected from either of polycarbonate, polymethylmethacrylate, and cyclic olefin copolymer.

[0034] The device 100 is pre- stored with all the required reagents or buffer solutions. In addition, the device 100 is provided with a lid (not shown in FIG.l). The lid is fixed on the first side S 1 of the device 100 to seal the first, second, and third air inlets and to seal the first and second air outlets. In an example, the lid is to be peeled-off before using the device 100. The lid is to prevent the damage or the leakage of the reagents or buffer solutions before use of the device 100. In an example, the device 100 is formed a thermoplastic material or from either of a polypropylene material, polycarbonate material, polymethylmethacrylate material, and cyclic olefin copolymer material.

[0035] The device 100 is formed such that an air-tight sealing is maintained throughout that is suitable for performing the pneumatic operations.

[0036] Although the device 100 depicted in FIG. 1 has a quadrilateral structure, the structure may vary according to the requirements, for e.g., synchronization or compatibility with the external optical detection system with different configurations. In an example, the device 100 may have a triangular structure.

[0037] FIG. 2A illustrates an isometric view of the device 100 of FIG. 1 for extracting Ribonucleic Acid (RNA) from a cell sample, according to an example implementation of the present subject matter. The quadrilateral structure of the device 100 is composed of two portions i.e., an upper portion 202 and a lower portion 204. The lower portion 204 of the device 100 is shown more clearly in FIG.2B, which illustrates an isometric view of the device 100 having the upper portion 202 and the lower portion 204 separated from one another. In an example, the upper portion 202 is joined with the lower portion 204 by performing an ultrasonic bonding process. In an example, the ultrasonic bonding process is one of ultrasonic welding, laser welding, and heat welding. In an example, the upper portion 202 is joined with the lower portion 204 by a double-sided tape. In an example, the upper portion 202 is joined with the lower portion 204 by a glue.

[0038] The upper portion 202 of the device 100 is injection molded. The lower portion 204 of the device 100 is flat. The lower portion 204 of the device 100 includes guidelines 206 on which the upper portion 202 is joined so as to make the joining portions airtight. In an example, the lower portion 204 of the device 100 is made with optically transparent material for optical readout to monitor the progression on nucleotide amplification. In an example, the upper portion 202 of the device 100 is made with optically transparent material for optical readout to monitor the progression on nucleotide amplification. In an example, the device 100 is made with optically transparent material for optical readout to monitor the progression on nucleotide amplification.

[0039] A length of the device 100 is in the range from 80 millimetres to 100 millimetres. In an example the length of the device 200 is 90 millimetres. A width of the device 100 is in the range from 40 millimetres to 50 millimetres. In an example the width of the device 200 is 45 millimetres. A height of the device 100 is in the range from 6 millimetres to 8 millimetres. In an example the height of the device 200 is 7.1 millimetres.

[0040] In addition, the upper portion 202 of the device 100 has a window

208. The window 208 is formed to expose the analysis unit 140 to the external optical detection system and also to lock the device 100 with the external optical detection system while in an operating mode related to detection of RNA. The window 208 is placed on the upper portion 202 such that a total height of the device is increased in a range from 10 millimetres to 12 millimetres. In an example the total height is 11.1 millimetres. [0041] Further, the lower portion 204 of the device 100 is provided with three alignment pins (not shown) and the upper portion 202 of the device is provided with three receptacles (not shown). The receptacles are formed in-line with the alignment pins and are capable of receiving the alignment pins. This reception is symbolic of a male and female connection. The male and female connection is to keep the upper portion 202 and the lower portion 204 of the device 100 intact with each other. Although, three sets of the alignment pins and the receptacles are provided in the device 100 as depicted in FIG. 2, the number of sets of the alignment pins and the receptacles may vary.

[0042] Furthermore, the device 100 has a slot (not shown) for receiving an identification unit. The slot is formed on the upper portion 202 of the device 100 right above a space, where the first storage chamber (L) (as shown in FIG.l) is accommodated in the device 100 of FIG. 1. Although the slot is explained to be formed on the upper portion 202 of the device 100, in another example, the slot is formed on the lower portion 204 of the device 100. The location of the slot is defined such that detection of the released RNA by the external optical detection system is not disturbed. The identification unit is to record details of a patient, of which the sample has been collected. In an example, the identification unit is a barcode chip. In another example, the identification unit is a QR code chip. The device 200 is used in combination with an external scanner to scan the identification unit for storing the details of the patient in a required storage server, such a hospital database.

[0043] FIG. 3A illustrates an isometric view, a top view and a front view of the extraction unit 122 of the device 100 of FIG. 1, according to an example implementation of the present subject matter. The extraction unit 122 has a volume of about 79 cubic millilitres. The dimensions of the extraction unit 122 are measured in millimetres and are clearly marked in FIG.3A. FIG. 3B illustrates an isometric view, a top view and a front view of the extraction unit 122 of the device 100 of FIG. 1, according to another example implementation of the present subject matter. The dimensions of the extraction unit 122 as per another example are measured in millimetres and are clearly marked in FIG.3B. FIG. 3C illustrates an isometric view of the extraction unit 122 having the filter 124.

[0044] FIG. 4 illustrates an isometric view, a top view and, a front view of the extraction unit 122 connected with the analysis unit 140 of the device 100 of FIG. 1, according to an example implementation of the present subject matter. The dimensions of the extraction unit 122 and the analysis unit 140 and their dimensional relationship with each other are measured in millimetres and are clearly marked in FIG.4.

[0045] FIG. 5 illustrates an isometric view, a top view and a front view of the waste chamber 144 of the device 100 of FIG.l, according to an example implementation of the present subject matter. The waste chamber 144 has a volume of about 2.2 millilitres. A length of the waste chamber 144 is in the range from 20 millimetres to 30 millimetres. In an example the length of the waste chamber 144 is 27.6 millimetres. A width of the waste chamber 144 is in the range from 11 millimetres to 15 millimetres. In an example the width of the waste chamber 144 is 14.4 millimetres. A height of the waste chamber 144 is in the range from 7 millimetres to 9 millimetres. In an example the height of the waste chamber 144 is 8 millimetres.

[0046] Although examples for the present disclosure have been described in language specific to structural features and/or methods, it is to be understood that the appended claims are not limited to the specific features or methods described herein. Rather, the specific features and methods are disclosed and explained as examples of the present disclosure.

Claims

We claim:

1. A device (100) for extracting Ribonucleic Acid (RNA) from a cell sample, the device comprising: a first storage unit L (118) having a lysis reagent (Rl) pre-stored therein, wherein the first storage unit L (118) is to receive the cell sample for mixing with the pre-stored lysis reagent (Rl) for lysis thereof; an extraction unit (122) having a first inlet (126), a second inlet (128), a third inlet (130), a first outlet (134), and a filter (124), wherein the extraction unit (122) is to: receive the lysed cell sample from the first storage unit L (118) via the first inlet (126) connected to the first storage unit L (118), wherein the filter (124) is to collect the lysed cell sample and to bind RNA out of the lysed cell sample; receive a wash reagent (R2) via the second inlet (128) to wash out inhibitors from the RNA, wherein the wash reagent (R2) is pre-stored within the device (100); and receive an elusion reagent (R3) via the third inlet (130) to elude the washed RNA from the filter (124), wherein the elusion reagent (R3) is pre stored within the device (100); and an analysis unit (140) connected to the first outlet (134) of the extraction unit (122), wherein the analysis unit (140) comprises a plurality of dried membranes (146) for nucleotide amplification, wherein the analysis unit (140) is to receive the eluded RNA from the extraction unit (122) and to divert the eluded RNA to each of the plurality of dried membranes (146), wherein each dried membrane (146) comprises an amplification reagent.

2. The device (100) as claimed in claim 1, wherein the analysis unit (140) comprises three dried membranes (146).

3. The device (100) as claimed in claim 1, wherein the device (100) comprises a first air inlet (102), a second air inlet (104), a third air inlet (106), a first air outlet (108), and a second air outlet (110), wherein the first air inlet (102), the second air inlet (104), and the third air inlet (106) are to receive a positive pressure, and wherein the first air outlet (108) and the second air outlet (110) are to receive a negative pressure.

4. The device (100) as claimed in claim 3, wherein the device (100) comprises a waste chamber (144) connected to a second outlet (136) of the extraction unit (122) and to the first air outlet (108), wherein the waste chamber (144) is to collect the lysed cell sample and residual reagents after the RNA binds to the filter (124), when the third air inlet (106) connected to the first storage unit L (118) receives the positive pressure and the first air outlet (108) receives the negative pressure.

5. The device (100) as claimed in claim 4, wherein the device (100) comprises a second storage unit W (150) having the wash reagent (R2) stored therein and connected to the first outlet (134) of the extraction unit (122) and to the first air inlet (102) of the device (100), wherein the second storage unit W (150) is to supply the wash reagent (R2) to the extraction unit (122), when the first air inlet (102) receives the positive pressure and the first air outlet (108) receives the negative pressure.

6. The device (100) as claimed in claim 3, wherein the device (100) comprises a third storage unit E (154) having the elusion reagent (R3) stored therein and connected to the third inlet (130) of the extraction unit (122) and to the second air inlet (104) of the device (100), wherein the third storage unit E (154) is to supply the elusion reagent (R3) to the extraction unit (122), when the second air inlet (104) receives the positive pressure and the second air outlet (110) connected to the diversion unit (140) receives the negative pressure.

7. The device (100) as claimed in claim 3, wherein each of the first air inlet (102), the second air inlet (104), the third air inlet (106), the first air outlet (108), and the second air outlet (110) comprise a glue for sealing thereof in an air-tight manner.

8. The device (100) as claimed in claim 7, wherein the glue is a thermoplastic polyurethane glue.

9. The device (100) as claimed in claim 7, wherein the glue is pierceable.

10. The device (100) as claimed in claim 1, wherein the device (100) comprises a passage (112) having one end connected to the first storage unit L (118) and another end having a first opening (114) to receive a cell sample collection device.

11. The device (100) as claimed in claim 1, wherein the device (100) comprises an upper portion (202) and a lower portion (204), wherein the lower portion (204) is made of an optically transparent material.

12. The device (100) as claimed in claim 1, wherein the device (100) is injection moldable.