Hygiene monitoring device, electronic optical reader apparatus and related method

Abstract

The present invention relates to a monitoring device for monitoring the hygienic status of a cannulated instrument, such as an instrument used in medical procedures, said device being capable of detecting proteins, an electronic optical device for carrying out an optical signal-generating reaction for the detection and quantification of proteins, and a method for monitoring the presence of proteins in a cannulated instrument.

Description

[0001] HYGIENE MONITORING DEVICE, ELECTRONIC OPTICAL READER APPARATUS AND RELATED METHOD

[0002] TECHNICAL FIELD

[0003] The present invention relates to a monitoring device for monitoring the hygienic status of a cannulated instrument, such as an instrument used in medical procedures, said device being capable of detecting proteins, an electronic optical device for carrying out an optical signal-generating reaction for the detection and quantification of proteins, and a method for monitoring the presence of proteins in a cannulated instrument.

[0004] BACKGROUND

[0005] Proper hygiene of cannulated instruments, and more particularly medical cannulated instruments, poses challenges within healthcare environments due to the complexity of their designs. Cannulated instruments frequently possess narrow, elongated cavities and intricate angles that hinder access during cleaning processes, creating an elevated risk of retaining biological material such as blood, tissue, and other body fluids.

[0006] Numerous studies indicate that inadequately cleaned instruments can lead to nosocomial infections and additional complications, as residual biological waste may persist post-cleaning and disinfection. Of particular concern are protein residues, which may act as vectors for transmitting prion diseases, such as variant Creutzfeldt-Jakob disease (vCJD), while also shielding bacteria and viruses from effective sterilization, allowing them to survive and pose risks to patient safety.

[0007] While advancements have been made in cleaning and disinfection technologies, current devices and methods for monitoring the cleanliness of cannulated instruments fall short in accurately detecting residual proteins. This insufficiency heightens the risk of crosscontamination, exposing patients to harmful pathogens. Given the critical importance of ensuring medical instrument cleanliness to prevent healthcare-associated infections, there is an urgent need for devices capable of reliably detecting and / or quantifying protein residues after cleaning.

[0008] PRO1 Micro Hygiene Monitoring System (hereinafter “PRO1 Micro”), is a device developed by TERRAGENE for the detection of proteins on surfaces. Using a swab, a technician is able to take a sample on a flat surface, and then the sample-loaded swab is introduced in the PRO1 MICRO Hygiene Monitoring System. Inside the device, the sample is contacted with a reactive solution that produces a change in color that can be measured. The concentration of proteins can then be determined by the colorimetric change of the reaction.

[0009] However, the PRO1 Micro device is not designed for the proper detection of samples in cannulated instruments, as evidenced by the type of swab utilized for sampling, and further does not comprise a mechanism to enhance the flow of a reagent from a chamber designed for the preservation of a reagent into another chamber wherein a reaction is meant to take place.

[0010] BRIEF DESCRIPTION OF THE INVENTION

[0011] In view of the limitations of the state of the art, the inventors provide herein a description of a device and method to carry out the analysis of the hygienic status of cannulated instruments, having improved features not only fortaking samples but also for preserving the reactants.

[0012] In a first aspect, the present invention provides a device for monitoring the hygienic status of a cannulated instrument, wherein the device comprises:

[0013] a reading chamber at one end of the device,

[0014] a reagent chamber placed contiguously to the reading chamber and comprising two frangible membranes each located at opposite ends of the reagent chamber, a channel chamber placed contiguously to the reagent chamber such that the reagent chamber is placed between the reading chamber and the channel chamber, and wherein the channel chamber comprises

[0015] a channel tube having a cutting end for breaking the frangible membranes, placed within the channel chamber, and

[0016] a piston-like cap at the other end of the device.

[0017] In an embodiment, the reagent chamber comprises a reagent composition having • a dye suitable for protein detection, such as a triphenylmethane dye, preferably, a dye with affinity for proteins such as Coomasie Blue G, at a concentration of at most 2% w / w; e.g., between 0-2% w / w, preferably between 0.01-2% w / w,

[0018] • a diluent suitable for a dye comprising an alcohol group, preferably in a concentration of at most 98% w / w.

[0019] In another embodiment, the reagent chamber comprises a reagent composition having • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninicacid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0020] • an oxidant capable of reacting with a protein such that allows the formation of a colored complex with the chelating agent,

[0021] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, at a concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0022] • water at a concentration of at most 80% w / w,

[0023] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w.

[0024] In another embodiment, the reagent chamber comprises a first reagent composition having

[0025] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / , preferably between 0.01-4% w / w,

[0026] • water at a concentration of at most 96% w / w;

[0027] and the reading chamber comprises a second reagent composition having

[0028] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninic acid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0029] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, ata concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0030] • water at a concentration of at most 80% w / w.

[0031] In an embodiment, the cutting end of the channel tube comprises a beveled edge.

[0032] It is a third aspect of the invention to provide a method for monitoring the hygienic status of a cannulated instrument comprising the steps of

[0033] a) taking a sample from the cannulated instrument,

[0034] b) introducing the sample in a device of the first aspect of the invention, c) contacting the sample with a reagent composition to perform an optical signalgenerating reaction, d) determining the concentration of protein present in the sample.

[0035] In an embodiment, the optical signal-generating reaction of step b) is performed at a temperature between 55-65°C, for a period of 1-10 minutes, and the absorbance is measured at 550-580 nm.

[0036] In an embodiment, the optical signal-generating reaction comprises a colorimetric reaction or a fluorescence reaction.

[0037] In an embodiment, the step d) is carried out by using a standard calibration curve.

[0038] BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 shows a schematic lateral view of an exemplary embodiment of the device of the first aspect.

[0040] Figure 2 shows a schematic lateral view of the components of an exemplary embodiment of the device of the first aspect.

[0041] Figure 3 shows a schematic representation of different embodiments of the channel tube comprised by the monitoring device.

[0042] Figure 4 shows an exemplary absorbent swab used in a particular embodiment of the device of the present invention.

[0043] Figure 5 shows a standard calibration curve of absorbance vs. bovine serum albumin (BSA) levels of an embodiment of a reagent solution. The absorbance is measured at 562 nm after 4 minutes of incubation at 60°C.

[0044] Figure 6 shows a series of standard curve calibrations of absorbance change vs BSA levels at three temperatures (40°C, 50°C, 60°C) and with an incubation time of four minutes.

[0045] Figure 7 shows a schematic representation of an exemplary mechanism of action of the device of the present invention, showing the breaking of the frangible membranes -represented with X - as the cutting edge of the channel tube penetrates these. A) An initial position of the device with the membranes intact and the channel tube in its resting position. B) An intermediate position wherein one of the membranes has been punctured by the cutting end of the channel tube. C) A final position wherein the two membranes have been punctured by the cutting end of the channel tube.

[0046] Figure 8 shows an embodiment of an automatic optical reader apparatus of second aspect, featuring one readout position 801 or well. User inputs and indication lights are possible through a membrane keyboard 802. The device is designed to allow for verification and calibration of the well temperatures using an external thermometer, which can be inserted through a dedicated calibration orifice. A protective cover 803 is incorporated to preserve the reading sensors while the device is unused.

[0047] Figure 9 shows another embodiment of an automatic reader apparatus featuring three independent readout positions or wells 901 and an integrated thermal printer 903 for registering results. User inputs and indication lights are possible through a membrane keyboard 902. A calibration thermometer can be inserted through a dedicated calibration orifice 904. A protective cover 905 is incorporated to preserve the reading sensors while the device is unused.

[0048] Figure 10 shows two embodiments of heating blocks. A) Shows an individual heating block for a single reading position. B) shows a heating block of a three-position reader. C) shows a bottom view of a single reading well, where 1001 indicates the cavity for the sensing device, 1002 indicates the channel for the emitting device, 1003 indicates the well orifice keyed to the reading chamber 201, and 1004 indicates the retention clip mechanism for the device of the first aspect. D) shows a top view of a reading well, illustrating the keyed design of the position in greater detail.

[0049] Figure 11 shows two embodiments of the reading hardware. A) A single emitter 1101 and a single RGB color sensor 1102 are positioned opposite each other, with a gap between them to accommodate the device of the first aspect. B) A set of three pairs of sensors 1103 and emitters 1104 are arranged, each sensor directly opposite its corresponding emitter, also with a gap between them to align with the device of the first aspect. In this configuration the distance between each sensor and emitter pair is consistently maintained across all three pairs.

[0050] DETAILED DESCRIPTION OF THE INVENTION Each and every embodiment of the aspects, devices, apparatus, methods, procedures or objects of the present invention resulting from the combination of particular embodiments described herein shall be considered as falling within the scope of the present invention.

[0051] Any technical and / or scientific terminology used herein shall be understood by the common definition utilized in the art and / or by those skilled in the art, unless otherwise explicitly stated or inferred by context.

[0052] As used herein, the terms “from X to Y”, or “between X and Y” or “from I between X-Y” and similar expressions, should be interpreted to include X and Y, unless the context indicates otherwise.

[0053] As used herein, the terms “a”, “an”, and “the” refer to “one or more” or “at least one” when used in this application, including the claims. Thus, for example, reference to “the hygienic status of an instrument” includes the hygienic status of a plurality of instruments, likewise reference to “detecting a protein” includes detecting a plurality of proteins, and so forth.

[0054] As used herein, the terms “comprise(s)”, “comprising”, “have”, “has”, “having” is intended to be inclusive and open-ended. It means “including, but not limited to”, and does not exclude additional, non-recited elements or steps. Thus, a system, device, method, or process that comprises one or more components or steps does not exclude the possibility of additional components or steps being present, even though they are not expressly listed.

[0055] As used herein, the term “embodiment”, and similar expressions like “an embodiment”, “one embodiment”, “a preferred embodiment”, etc., indicates the description of a particular embodiment of a device, method, or process of an aspect of the present invention that may include a particular feature, structure, or characteristic, but every embodiment may not necessarily include that particular feature, structure, or characteristic. Furthermore, the expression may not necessarily refer to the same embodiment. When a particular feature, structure, or characteristic is associated or connected to a particular embodiment, it is submitted that is within the knowledge of the person skilled in the art to implement such feature, structure, or characteristic in connection with other embodiments, whether or not explicitly described. As used herein, the term “hygienic status”, in particular in the context of an instrument or tool, refers to the condition in which said instrument or tool is in relation to the presence of proteins, such as residual proteins or undesired proteins. That is, the term “hygienic status” refers mainly to the presence or absence of residual proteins, and secondarily to the quantity of said residual proteins when present. The hygienic status may be qualitatively defined as “clean” or “not clean” depending on whether there is a detectable level of proteins present in said instrument. Said status can be previously determined by the user. Alternatively, additional categories can be defined qualitatively or quantitatively, and these are preferably defined in relation to the concentration of protein present or detected in the instrument measured in any type of scale, e.g., a linear or logarithmical scale.

[0056] As used herein, the term “cannulated instrument” or “cannulated tool” refers to an instrument designed with a hollow core, cannula or channel running through its length. In particular, the term may refer to cannulated instruments for use in medical or surgical procedures, for example a cannulated medical device.

[0057] As used herein, the term “protein” refers to a polypeptide molecule or molecules that can be considered a residue or which presence is considered undesired in an instrument, for example a cannulated instrument. The term refers to polypeptide molecules that can be detected and quantified by any means available in the state of the art, for example, by means of an optical signal-generating reaction, such as a colorimetric or a fluorescence reaction. The term may also refer to a single type of protein, ora mixture of proteins. The protein may be present in an isolated state, or as part of a structure, like a subcellular or cellular residue, or as part of a cell.

[0058] Throughout the present application, the terms “chamber” and “compartment” will be used interchangeably, and refer to a hollow structure that is part of a device and that performs a certain function, or wherein a certain function is performed.

[0059] As used herein, the term “frangible” refers to the quality of a material or element to break under desired conditions. A frangible material or element, e.g., a membrane, thus is designed to maintain its structural integrity until it is intentionally sought to be destroyed, for example, by applying a considerable force or pressure such as that of a cutting or perforating element. It is a first aspect of the present invention to provide a device for monitoring the hygienic status of a cannulated instrument, wherein said device comprises:

[0060] • a reading chamber at one end of the device,

[0061] • a reagent chamber placed contiguously to the reading chamber and comprising two frangible membranes each located at opposite ends of the reagent chamber, • a channel chamber placed contiguously to the reagent chamber such that the reagent chamber is placed between the reading chamber and the channel chamber, and wherein the channel chamber comprises

[0062] o a channel tube having a cutting end for breaking the frangible membranes, placed within the channel chamber, and

[0063] • a piston-like cap at the other end of the device.

[0064] In an embodiment, the device can be used to monitor the hygienic status of a cannulated instrument selected from endoscopes, cannulated needles, and trocars.

[0065] The device determines the hygienic status of a cannulated instrument by detecting proteins that are present in a sample taken from a cannulated instrument. When present in a sample, the protein may be in an isolated state, or as part of a structure, like a subcellular or cellular residue, or as part of a cell.

[0066] The following is a detailed description of an embodiment of the device of the first aspect based on the elements shown in the accompanying Figures, in order to provide a description thereof. Therefore, the following description shall be interpreted only as an exemplary embodiment of the device provided by the present invention and is not intended to be limiting of the invention in any way. Those skilled in the art will recognize or be able to determine by routine experimentation many equivalents of the specific embodiments described herein. Such equivalents are considered to fall within the scope of the present invention.

[0067] It is a first aspect of the present invention to provide, as seen in Figs. 1 and 2, a device 200 for monitoring the hygienic status of a cannulated instrument, wherein said device 200 comprises:

[0068] a reading chamber 201 at one end of the device 200,

[0069] a reagent chamber 203 placed contiguously to the reading chamber 201 and comprising two membranes 209a and 209b each located at opposite ends of the reagent chamber 203, a channel chamber 206 enclosing the reagent chamber 203, the reading chamber 201 being located within or adjacent to said channel chamber 206, wherein said channel chamber 206 comprises

[0070] a channel tube 207 having a cutting end 301 for breaking the frangible membranes 209a and 209b placed within the channel chamber 206, and a piston-like cap 208 at the opposite end of the device 200.

[0071] The device comprises a linking connector 205 that connects the reagent chamber 203 and the channel chamber 206.

[0072] Furthermore, a first O-ring 202 is placed between the reading chamber 201 and the reagent chamber 203, while a second O-ring 204 is placed between the reagent chamber 203 and the linking connector 205. The O-rings ensure a secure fitting of the elements connected by them.

[0073] The reading chamber 201 comprises an open volume, said volume being sufficient for performing an optic signal-generating reaction triggered by the contact of a reagent composition and a protein under certain reaction conditions (e.g. temperature, pH, salinity). For this purpose, the reading chamber 201 is made of a transparent material suitable for performing and measuring an optic signal-generating reaction inside thereof. Said transparent material comprises copolymers selected from PETG, PCTA, and PCTG.

[0074] It is understood that, when a reaction takes place, the reading chamber 201 is filled partially or completely with a reagent composition and a sample that may or may not contain a protein. Throughout the reaction, changes in the emitting wavelength of the reacting mixture comprising the reagent composition and the sample can be detected depending on the presence and quantity of protein in the sample, and said changes can be quantified.

[0075] The optic signal-generating reaction that takes place inside the reading chamber 201 can be measured on any kind of instrument designed for such end, for example, an spectrophotometer, a colorimeter, or an electronic optical reader apparatus. In a preferred embodiment, the optic signal-generating reaction is measured in an electronic optical reader apparatus. Said apparatus can be configured to receive the reading chamber 203, incubate the reading chamber 203 at a predetermined temperature and time necessary for performing an optic signal-generating reaction, and measure any wavelength absorption changes that can take place inside the reading chamber 201.

[0076] An electronic optical reader apparatus, thus, comprises a reading ch amber- receiver, a calibrated light source for precise illumination at different wavelengths, an RGB photodetector capable of detecting changes in the absorbance of wavelengths, and at least one microprocessor or microcontroller to control the components. Optionally, the apparatus may include software with specific algorithms for processing the readings and interpreting the results.

[0077] Changes in the wavelength absorption of the mixture can result in variations in absorbance values of the mixture at different wavelengths that can be registered by the electronic optical reader apparatus. Preferably, the changes in absorbance are measured in wavelengths of 300-1000 nm, preferably between 400-700 nm, more preferably between 550-580 nm. For example, and without intent of limiting the scope of the present invention, for a colorimetric reaction, absorbance can be measured in wavelength ranging from 550-580 nm, while for a fluorescence reaction, absorbance can be measured in wavelength between 350-400 or between 430-520 nm.

[0078] Furthermore, as a person skilled in the art would appreciate, in orderto adapt the present aspects of the invention as to perform a fluorescence reaction, it would be necessary to modify the reagent composition by including a fluorescence-generating compound that reacts, i.e. generates fluorescence, in the presence of a protein. The wavelength absorbance thus would require to be between 350-530 nm, for example 250-400 nm or between 430-520 nm.

[0079] For example, and without intent to limit the scope of the present invention, the electronic optical reader apparatus may perform an incubation at 40-70°C, preferably 60°C; during a period of 1-10 minutes, preferably 4 minutes. Followed by an absorbance reading at wavelengths of 550-580 nm, preferably at 562 nm.

[0080] As evidenced in Fig. 5 provided herein, the level of absorbance has a linear relation with the concentration of a protein when measuring the absorbance of 562 nm. In the exemplified case, the linear relation is maintained between 0.5-60 pg of a reference protein (BSA) which can be used as a standard calibration curve of absorbance for the determination of concentration of unknown concentrations of protein present in a sample. The determination of protein concentration in a sample by comparison with a standard concentration is only an exemplary embodiment of how the device provided by the present invention can be used to determine the presence and concentration of proteins. This example is not meant to be limiting in any way, since other methods for determining the presence or concentration of proteins can be adapted by a person skilled in the art. It shall be understood that such methods fall within the scope of the present invention. A standard calibration curve can be obtained using any protein suitable for such end. A non-limiting example comprises using a bovine serum albumin (BSA) as the standard protein. In an embodiment, the standard calibration curve can be used to determine the concentration of proteins between 0.5-70 pg.

[0081] The reagent chamber 203 is designed to contain a reagent composition formulated for the detection of proteins, more specifically, the detection of proteins through an optic signal-generating reaction wherein a change in wavelength absorption can be detected. Additionally, the reagent chamber 203 keeps the reagent composition unaltered until the moment of use.

[0082] It falls within the knowledge of the person skilled in the art to optimize the formulation of the reagent composition in order to detect a particular type of protein, or to carry out the reaction in the way to be considered most suitable, e.g., to obtain a particular wavelength or to carry out the reaction at a specific pH, salinity, temperature, or incubation time, among other conditions. Non-limiting examples of colorimetric reactions include but are not limited to colorant-protein reactions or Copper-protein colorimetric reactions.

[0083] As used herein, the term “reagent composition” refers to a mixture of compounds, formulation, solution, etc., that reacts when in contact with a protein. In the context of the present invention, the reaction between the reagent composition and the protein is revealed by the emission of an optical signal, i.e., the resulting reaction between the reagent composition and a protein generates a modification in the spectrum of light that’s either absorbed, emitted, or both, and said modification can be measured.

[0084] In this context, the term “light” has to be understood in a broad sense and not limited to the spectrum of the electromagnetic radiation that is known as “visible light”, rather the spectrum that goes from 100 to 2000 nm.

[0085] In an embodiment, the reagent composition comprises a “pre-activated” reagent composition, i.e., a reagent composition that is ready to initiate a reaction upon contacting a sample comprising a protein under certain reaction conditions are met (e.g. temperature, pH, salinity, incubation time).

[0086] It is understood that the pre-activated reagent composition comprises all the components necessary for performing an optical signal-generating reaction when in contact with a sample that may or may not comprise a protein. Thus, in an embodiment the preactivated reagent comprises the compounds necessary to perform a colorimetric reaction or a fluorescence reaction.

[0087] In an embodiment, the pre-activated reagent composition comprises ...

[0088] • a dye suitable for protein detection, such as a triphenylmethane dye, preferably, a dye with affinity for proteins such as Coomasie Blue G, at a concentration of at most 2% w / w; e.g., between 0-2% w / w, preferably between 0.01-4% w / w,

[0089] • a diluent suitable for a dye comprising an alcohol group, preferably in a concentration of at most 98% w / w.

[0090] In another embodiment, the pre-activated reagent composition comprises

[0091] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninicacid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0092] • an oxidant capable of reacting with a protein such that allows the formation of a colored complex with the chelating agent,

[0093] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, at a concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0094] • water at a concentration of at most 80% w / w,

[0095] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w.

[0096] In another embodiment, the reagent chamber 203 comprises a first “non-activated” reagent composition and the reading chamber 201 comprises a second “non-activated” reagent composition. Each one of the non-activated reagent compositions is unable to initiate a reaction when in contact with a sample on their own. Thus, in this embodiment, in order to initiate a reaction, both the first and second non-activated reagent compositions have to be mixed before or during contact with a sample. In this embodiment, the mixture of the first and second “non-activated” reagent results in a “post-activated” reagent composition that comprises all the components necessary for performing an optical signal-generating reaction when in contact with a sample. Thus, in an embodiment the post-activated reagent comprises the compounds necessary to perform a colorimetric reaction or a fluorescence reaction.

[0097] In an embodiment, the post-activated reagent composition comprises

[0098] • a dye suitable for protein detection, such as a triphenylmethane dye, preferably, a dye with affinity for proteins such as Coomasie Blue G, at a concentration of at most 2% w / w, e.g., between 0-2% w / w , preferably between 0.01-2% w / w;

[0099] • a diluent suitable for a dye comprising an alcohol group, preferably in a concentration of at most 98% w / w.

[0100] In another embodiment, the post-activated reagent composition comprises

[0101] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninicacid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0102] • an oxidant capable of reacting with a protein such that allows the formation of a colored complex with the chelating agent,

[0103] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, at a concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0104] • water at a concentration of at most 80% w / w,

[0105] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w.

[0106] As a person skilled in the art would understand, the different components of the “postactivated” can be divided between a first and a second non-activated reagent compositions, and these components remain separated in reagent chamber 203 and the reading chamber 201, until reaction is needed. In an embodiment, the first non-activated composition comprises...

[0107] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0108] • water at a concentration of at most 96% w / w;

[0109] In an embodiment, the second non-activated composition comprises...

[0110] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninic acid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w,

[0111] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, ata concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0112] • water at a concentration of at most 80% w / w.

[0113] The reagent chamber 203 comprises a first membrane 209a placed between the reading chamber 201 and the reagent chamber 203, and a second membrane 209b placed between the reagent chamber 203 and the linking connector 205. These membranes seal the reagent chamber 203 ensuring that the reagent composition is kept undisturbed and under preservation conditions until used. Furthermore, both these membranes are made of a frangible material. These frangible membranes function as a controlled activation system, impeding the introduction of the reagent composition into the reading chamber until a sample analysis is required. This being said, the reagent composition can be contained in preservation conditions up to 2 years.

[0114] As such, the frangible membranes are made of a frangible material. The frangible material can be of any material with enough resilience to avoid accidental spills or leaks of the reagent composition, but weak enough for being easily broken, e.g. punctured, by the cutting end 301 of the channel tube 207 in any of its embodiments (see, e.g., Fig. 3). Said frangible material is preferably selected from PET, PP, and a mixture of PET and PP.

[0115] The channel chamber 206 comprises an internal volume wherein a channel tube 207 designed to hold a swab 400. The channel chamber 206 can also be used for storing a sample that has been taken from a cannulated instrument, wherein said sample is adsorbed in the swab 400, preferably in the swab-head 401. As seen in more detail in Figure 3, the channel tube 207 consists of a tube comprising i) a cutting end 301 having a cutting edge and an opening,

[0116] ii) an insertion end 302 having an opening,

[0117] iii) an inner diameter 303 having enough space for allowing the passage of a swab 400, and

[0118] iv) optionally, a conical funnel 304.

[0119] The cutting end 301 has a cutting edge designed to ensure that the channel tube 207 can break the frangible membranes located at each end of the reagent chamber 203, thus allowing the contained composition to flow into the reading chamber 201. The cutting end 301 also comprises an opening.

[0120] The cutting end 301 may be present in different shapes and edges. For example, the cutting end comprises a beveled edge. Preferably a beveled edge of a shape selected from a J-bevel, V-bevel, Compound bevel, Compound J-Bevel, or any of these with or without counterbore. In another embodiment, the beveled edge can cut at an angle of 20°-30°. The beveled edge can comprise a hollow bevel with a central groove that enhances the reagent composition transfer from the reagent chamber 203 to the reading chamber 201.

[0121] Figure 3 shows exemplary embodiments of the channel tube 207. Tube 207a represents a channel tube with a beveled V-shape edge. Tube 207b represents a channel tube with an arrowhead-shaped edge with a central groove to enhance solution transfer. Tube 207c represents a channel tube with a multi-faceted bevel.

[0122] The insertion end 302 may be wide enough to allow for the proper insertion of a swab 400 into the inner diameter 303. In order to ensure the introduction of swabs of different sizes, the channel tube 207 comprises a conical funnel 304 that guides and facilitates the introduction of swabs of different sizes into the inner diameter 303. In order to secure the swab introduction, the diameter of the channel tube 207 narrows as it gets closer to the cutting end 301, e.g., the channel tube 207 has a funnel-like shape, with the cutting end 301 being narrower than the insertion end 302.

[0123] The length of the channel tube 207 is enough to completely cross both ends of the reagent compartment 203. In a preferred embodiment, the channel tube 207 measures between 1.00-5.00 cm and the reagent chamber 203 measures between 1.00-5.00 cm. Preferably, the ratio between the channel tube 207 length and the reagent chamber 203 length is of between 1.00-5.00. As seen in Fig. 4, the swab 400 comprises a swab-head 401 and an elongated tube or swab-body 402. The swab-head 401 is optimized for the efficient collection and retention of protein residues. The swab head can be moistened before use or at moment of use for improved the collection of samples. In a preferred embodiment, the swab-head 401 is moisturized with a surfactant comprising a nonionic surfactant, preferably selected from NP-40 (Nonidet P-40), and Tween 20 (Polysorbate 20). The moisturizing process enhances protein recovery by minimizing variations due to sample retention, ensuring reliable detection and / or quantification in conjunction with the automated optical reader apparatus.

[0124] The swab is made with a flexible material, and thanks to the elongated shape of the swab-head 401 and the narrow swab-body 402 (see Fig. 4), the swab is designed to be inserted in narrow cavities, making it ideal for picking up samples from cannulated instruments. In an embodiment, the swab-head 401 has a width of 1.0-4.0 mm, or a length of 1-2500 mm, preferably the swab-head 401 has a width of 1.7-3.0 mm and a length of 2000-2500 mm. In an embodiment, the swab-body 402 has a width of 0.5-3.0 mm, or a length of 2000-2500 mm, preferably, the swab-body 402 has a width of 0.7-1.5 mm and a length of 2000-2500 cm.

[0125] Additionally, the material used for constructing the swab is highly absorbent, increasing the efficiency of capture of residues that might be present in a cannulated instrument. Thus, in a preferred embodiment, the swab-head is made of synthetic polyamide fibers and the swab-body is made of nylon fiber.

[0126] Thanks to the materials used for the construction of the swab, it is possible to pick up samples from cannulated instruments that other types of swab cannot obtain, thus the swab that forms part of the device of the first aspect can have the intended measurements representing a technical advantage before other similar devices like PRO1 Micro, and provides a solution to the technical problem of performing hygienic monitoring of cannulated instruments.

[0127] In order to activate the device, the user presses the piston-like cap 208 in a way that it lodges into the channel chamber 206 until it reaches a blocking top located at the end of said tab 208. This causes the tab 208 in turn to push the channel tube 207 into the reagent chamber 203, breaking first the membrane 209b, and later causes to break the membrane 209a. By breaking both the membrane 209a and 209b, the channel tube 207 pierces through the reagent chamber 203, and in the process the reagent composition is introduced into the channel tube 207 through the opening of the cutting end 301 and contacts the sample contained in the swab 400 that may or may not contain a protein. As a result, the reagent composition once in the reagent chamber 203 now is free to flow into the reading chamber 201, wherein an optical signal-generating reaction can take place. This process is represented in Fig. 7.

[0128] The swab 400 can be inserted in the channel tube 207 by removing the piston-like cap 208 and introducing said swab by the insertion end 302. This process is preferably done after the reagent composition in any of its embodiments has been introduced in the reading chamber 201. More specifically, the element that is introduced in the channel tube 207 is the swab-head 401 which comprises a sample that may or may not contain a protein. As a result, the sample located in the swab-head 401 gets in contact with the reagent composition that has entered the channel tube 207 through the cutting end 301 thus initiating the optical signal-generating process.

[0129] In order to avoid spilling of the reagent composition inside the device, the device shall be handled in an approximately vertical position with the reading chamber 201 placed at the bottom, thus ensuring the flow of the reagent composition into the reading chamber 201 by gravity.

[0130] Alternatively, in an embodiment, the channel tube 207 has a maximum width at the insertion end 302 such that matches or is approximately the same as that of the reagent chamber, thus impeding the flow of the reagent composition into the channel chamber 206.

[0131] It is a second aspect of the invention to provide an electronic optical reader apparatus.

[0132] The electronic optical reader apparatus is designed for the precise detection and quantification of protein levels and record keeping of the results. This reader is compatible with any embodiment of the device of the first aspect disclosed herein, enabling efficient detection, analysis and documentation of protein residues within complex internal surfaces. Thus, the electronic optical reader apparatus can be used for the precise detection and quantification of protein residues that are present in a sample, preferably a sample taken from a cannulated instrument. Two exemplary embodiments of the apparatus are depicted in Figures 8 and 9. These are only meant to represent preferred embodiments of the apparatus of the invention and are not meant to limit the scope of the invention in any way.

[0133] The apparatus can comprise an outer shell designed for protecting the apparatus and its components. The shell may be constructed with a casted polymer or plastic. Non-limiting examples of casted polymers or plastics include Acrylonitrile Butadiene Styrene (ABS), more particularly ABS selected from POLYLAC® PA-757 ABS Resin from CHI MEI CORPORATION and TOYOLAC™ 900 from Toray Plastics.

[0134] In an embodiment, a protective cover, as shown in 803 and 905, may be included to prevent dust and liquid intrusion into the reading wells 801 or 901, respectively, which could otherwise interfere with the apparatus’ intended operation. This cover can be made of a casted polymer or plastic and may be semi-transparent. Non-limiting examples of casted polymers or plastics include WONDERLITE® PC-175 or the WONDERLITE® PC-110 from CHI MEI CORPORATION.

[0135] In an embodiment, the apparatus comprises a user interface which may take form of a membrane keyboard 802 or 902. This interface allows the user to configure the apparatus using physical buttons and monitor the operational status through indicator lights, such as light-emitting diodes (LEDs). In some embodiments, the apparatus is designed to accept voice commands and communicate updates to the user through prerecorded audio or, alternatively, via audio generated by an onboard processing unit. Additionally, the apparatus may feature an integrated touch sensitive display or support wireless control through a separate device, for example, through a computer, laptop, cellphone, smartphone or any other device capable of running a proper software application.

[0136] Figure 8 shows an exemplary embodiment of the apparatus of the second aspect, comprising a single reading well 801 where a device of the first aspect of the invention can be placed to detect and quantify proteins present in a sample, preferably, a sample taken from a cannulated instrument. Upon completion of the readout process, the result can be reported to a user through a user interface as previously described. Additionally, the apparatus may include both wired or wireless connectivity options to transmit any result to an application or quality assurance system for further evaluation and regulatory compliance where applicable. In an embodiment, the connectivity module and optical module are handled separately by separate processing units.

[0137] Figure 9 shows an exemplary embodiment of the apparatus of the second aspect, comprising three readout position or wells 901. In this embodiment, athermal printer 903 can be integrated to provide printed results of the readout carried out on the monitoring device of the first aspect, ensuring compliance with applicable regulations and customer reporting requirements. Additional information that may not be displayable on the user interface can be included in a physical report.

[0138] Temperature plays a critical role in achieving the proper reaction conditions and kinetics in the device of the first aspect while detecting and quantifying the protein levels in a sample. Thus, it is a key aspect of the apparatus to heat and maintain a predefined temperature within the readout wells. The temperature of the reading positions can influence the dynamics and the variation of light absorption of the solution that may or may not contain protein residues, accelerating the reaction as the temperature increases. In some embodiments, the well temperature is heated to and maintained at a temperature of between 37-70°C, preferably between 50-65°C. more preferably between 58°C-62°C, for example 58, 59, 60, 61 or 62°C.

[0139] As shown in Figure 9, the apparatus comprises an orifice 904. Orifice 904 is designed for placing an external thermometer, preferably for calibration and verification of temperature. This orifice allows the external thermometer to make direct contact with each individual well 901 in embodiments where the device of the first aspect is heated individually, or to contact the entire heating block in embodiments where the reading wells 901 share a single heating block.

[0140] Properly heating the device of the first aspect is crucial for achieving a desired result and reaction kinetic. To control and maintain the temperature during readout, the apparatus can be equipped with any one selected from a heating block, a heating element, a temperature sensor, and a thermal protection.

[0141] Figure 10 A) shows an embodiment of the heating block corresponding to a single readout position, for an apparatus as depicted in Figure 8, e.g., comprising a single reading well. Figure 10 B) shows an embodiment of the heating block used in an apparatus illustrated in Figure 9, e.g., comprising three reading wells. In any of the embodiments, the heating block comprises a thermally conductive material, preferably a metal alloy. Non-limiting examples of alloys include aluminum alloys, e.g. ADC12, 6082, 6060, or 6061. These are particularly preferred due to their high thermal conductivity-to-weight ratio. The heating block can be either cast or machined.

[0142] The apparatus of the second aspect comprises a heating element. The heating element, comprises at least one electric resistor, wherein said at least one electric resistor is controlled by a feedback loop on the apparatus’ main microcontroller. Preferably, the heating element consists of at least one electric resistor. This loop operates through a switching element, such as a Field Effect Transistor (FET), and uses temperature measurements from a digital sensor, which are interpreted by the microcontroller to regulate heating to a predefined setpoint.

[0143] As shown in the corresponding Figures 10C and 10D, which correspondingly display the bottom and top a single reading well, the heating block not only provides heating for the device of the first aspect but is also keyed to match the shape of the reading chamber 201, as appreciated in the element 1003. This alignment ensures that the reading chamber 201 of the device of the first aspect is precisely positioned to face the sensing unit and the light emitter, respectively, maximizing the optical benefits of the device’s design. The cavity 1001 for the sensing unit and the channel 1002 are the compartments for the sensing unit and the emitter, respectively. A single circular orifice, positioned perpendicular to these compartments, is located at the exact height at which both the sensor and emitter align with a reading chamber 201, orto a device of the first aspect in any of its embodiments. This orifice prevents the passage of diffused or stray light, allowing only the light transmitted from the emitter and not absorbed by the device of the first aspect’s solution to reach the sensor.

[0144] 1004 represents a retention mechanism that can be present in some embodiments of the apparatus of the second aspect. Due to the physical characteristics of the device of the first aspect, it may not fit fully in the well or could tilt to one side, which could negatively impact the readout performance. The retention clip 1004 ensures firm and secure placement of the device of the first aspect, thus enabling precise and reliable readout.

[0145] The detection of protein in the device of the first aspect, resulting from the biochemical reaction produced after the reagent composition contact with the flexible swab used to sample a cannulated instrument at temperature conditions controlled by the heating block, is carried out by the optical system of the electronic optical reader apparatus.

[0146] As previously established, protein residue quantification is based on changes in light absorbance as the biochemical reaction progresses. The initial and final absorbance values can be used to indirectly estimate the protein levels present originally present in the sample. In some embodiments, a variable proportional to the rate of absorbance change — measured at a specific reaction temperature and wavelength — may be used along with a calibration curve to quantify protein residues on the reactive pen. The peak lightwavelength for the analysis is, preferably, (562 ± 20) nm, which aligns with the onset of the biochemical reaction. In the presence of protein residues, this reaction results in an increase in light absorption at this wavelength as it progresses.

[0147] The optical system consists of a calibrated light source and a sensing unit, with the reading chamber positioned between them at an optimized distance. An optical window is designed to maximize light capture from the reading chamber while blocking any extraneous light. The distance between the sensor and the reading chamber 201 , as well as between the sensor and light source, can be influenced by the physical characteristics of the electronic components, such as the angle of vision, angle of emission, sensor sensitivity, and luminosity.

[0148] Figure 11 shows exemplary embodiments for single position optical systems 1101, or a plurality of independent optical systems 1102. The sensing units and light sources can be mounted on printed circuit boards (PCBs) perpendicular to a main circuit board, and parallel to the reading chamber 201 , specifically parallel to the planar faces.

[0149] The light source may be one or more Light-Emitting Diodes (LEDs), fluorescent lamps, or lasers and it is directly controlled by the optical reading microcontroller. Preferably, the light source is one capable of emitting light in a range between 300-1000 nm. While any source capable of emitting light at 562 nm ± 20 nm would produce adequate results, a white LED is preferred. Examples of suitable white LEDs are the LTW-670DS(-EL), and the LTST-E683CEGBW from Lite-ON and the ASMT-QWBF-NKLOE from Broadcom Limited. This choice enables the apparatus to function not only with the biochemical solutions present in the reactive pen for cannulated instruments but also with other products where light transmission and absorbance techniques can be utilized for indirect detection and quantification of a magnitude of interest. In an embodiment, the white LED is the LTW-670DS and the distance between a white LED to the reading chamber 201 is between 10-20 mm, preferably 16 mm and its height is set at between 8.0 and 12.0 mm, preferably 9.5 mm, so it is aligned with the reactive solution inside the reading chamber. That is, the distance between the light emitter and the center of the reading well or the perpendicular orifice is between 10-20 mm, and its height is set at between 8.0 and 12.0 mm, preferably 9.5 mm.

[0150] The sensing unit comprises a sensor, a signal conditioning circuit, a microcontroller for signal processing, and optionally, an optical filter (typically a high pass or bandpass filter). The signal conditioning circuit is highly dependent on the sensor choice. In at least one embodiment, the sensor is a digital sensor and its signals can be directly interpreted by the microcontroller. In a preferred embodiment, the sensor is analog and a low-pass filter circuit and analog to digital circuit are implemented before the signal is interpreted by the microcontroller.

[0151] The sensor may be a wide-spectrum photodiode with a bandpass filter centered in 560 nm, or a monochromatic sensor like the S6429-01 from Hamamatsu or the EACLSST3227A1 from Everlight, designed specifically for green color detection (450 nm < Ap < 650 nm). Alternatively, the sensor can be multichannel, commonly referred to as color sensors or RGB sensors. The latter alternative is preferred for the same reasons exposed for the light emitter. Examples of suitable color sensors are the VEML3328 from Vishay Semiconductors and the S7505-01 or S9702 manufactured by Hamamatsu. In an embodiment, the sensor is the S7505-01 from Hamamatsu. The distance between the reaction well and the sensor can be between 3-25 mm, preferably 15 mm; and the center of the sensing unit is between 8.0-12.0 mm, preferably 9.5 mm, to align with the reading chamber 201.

[0152] Signal processing is performed by a dedicated microcontroller within the apparatus. Alternatively, a single microcontroller unit may handle configuration, communication with other devices, user interface and user experiencing, and processing functions. The apparatus also includes non-volatile memory, such as EEPROM or eMMC, dedicated to storing calibration parameters, user-defined thresholds for acceptable and alarm protein residue levels, and readout results.

[0153] Independent of the hardware configuration, the microcontroller responsible for signal handling and processing also controls the activation and deactivation of the light emitters, as well as their intensity. This is managed either through pulse-width modulation (PWM) or constant current control. The microcontroller also receives and processes the conditioned signals obtained from the analog-to-digital converter within the measurement circuit.

[0154] The typical readout process for a device of the first aspect for detecting and quantifying protein residues on a cannulated instrument involves collecting a sample from a cannulated instrument, activating the device of the first aspect with the swab to transfer protein residues into the reagent composition, selecting the appropriate readout program on the electronic reader apparatus, and initiating the readout process through the user interface. Once the device is positioned in the reading well, the heating block applies heat to facilitate the biochemical reaction.

[0155] In an embodiment of the apparatus, afterthe readout process is started by the indication of the user, the LED emitter is turned on and the processing unit takes samples from the measuring circuit at a regular interval between 100 and 700 milliseconds. In a preferred embodiment, the samples are taken every 500 milliseconds.

[0156] In another embodiment of the apparatus, afterthe user initiates the readout process, the actual readout may be delayed to allow the reaction kinetics to stabilize. In this case, the readout begins between 30 and 100 seconds afterthe initial start command.

[0157] A magnitude relative to the reaction kinetics in the presence of protein residues is calculated for detection and quantification. For instance, the rate of change in absorbance can be determined by calculating the instantaneous derivative of the signal captured by the color sensor as the biochemical reaction proceeds under illumination from the calibrated light source.

[0158] The results can be accumulated (e.g. a pondered sum) and a value can relative to the reaction kinetics can be obtained.

[0159] The resulting value can be further processed to detect and quantify protein residues. An experimental Bovine Serum Albumin (BSA) calibration curve can be established to correlate the values obtained by the processing unit with a BSA equivalent in micrograms. To develop this calibration curve, a series of experiments are conducted, allowing known quantities of BSA to react with the reactive pen, enabling accurate correlation between sensor output and protein residue concentration. Figures 5 and 6 shows a calibration curve and a series of calibration curves, respectively, for bovine serum albumin protein concentration. These values are recorded for BSA concentrations ranging from 0 to 70 pg over a 4-minute readout process, with each curve representing data at a single (Fig. 5) or different reaction temperatures (Fig. 6). The graphs indicate that the magnitude proportional to the change in absorption increases with temperature for a given BSA concentration, highlighting the impact of temperature on reaction kinetics and absorbance.

[0160] For a specific temperature setpoint, a calibration curve can be established, where the calibration parameters may be represented by a linear approximation, a logarithmic approximation, or a combination of both approaches. This allows for more accurate quantification of protein residues based on the observed changes in absorbance at the given temperature.

[0161] The obtained values can then be compared to predefined threshold values set by the user to determine the admissibility of protein residues found on the sample. In some embodiments, the results can be printed using an integrated thermal printer. Additionally, the apparatus may feature light indicators, such as LEDs, to indicate the result of the readout process (e.g., pass or fail).

[0162] Furthermore, the apparatus may have connectivity options to communicate with other electronic devices. This can include a wired connection through a USB port or wireless communication via Bluetooth or Wi-Fi. This allows the apparatus to share the readout state and results with other electronic systems for monitoring, digital record-keeping, or integration with larger quality assurance or regulatory systems.

[0163] It is a third aspect of the invention to provide a method for monitoring the hygienic status of a cannulated instrument comprising the steps of

[0164] e) taking a sample from the cannulated instrument,

[0165] f) introducing the sample in a device of the first aspect of the invention, g) contacting the sample with a reagent composition to perform an optical signalgenerating reaction,

[0166] h) determining the concentration of protein present in the sample.

[0167] The following is a detailed description of an embodiment of the method of the third aspect based on the elements shown in the accompanying Figures, in order to provide a description thereof. Therefore, the following description shall be interpreted only as an exemplary embodiment of the method of the present invention and is not intended to limit in any way. Those skilled in the art will recognize or be able to determine by routine experimentation many equivalents of the specific embodiments described herein. Such equivalents are considered to fall within the scope of the present invention.

[0168] The cannulated instrument from which the sample is taken comprises a cannulated instrument for use in medical or surgical procedures. Preferably, the method can be used to monitor the hygienic status of a cannulated instrument selected from endoscopes, cannulated needles, and trocars.

[0169] The sample can be taken by using a swab, for example, a swab like the one referred to herein as swab 400, comprising a swab-head 401 and a swab-body 402. The method comprises moisturizing the swab-head 401 prior to taking the sample. In a preferred embodiment, the swab-head 401 is moisturized with a surfactant comprising a nonionic surfactant, preferably selected from NP-40 (Nonidet P-40), and Tween 20 (Polysorbate 20).

[0170] In an embodiment, the swab-head 401 has a width of 1.0-4.0 mm, or a length of 1 -2500 mm, preferably the swab-head 401 has a width of 1.7-3.0 mm and a length of 2000-2500 mm. In an embodiment, the swab-body 402 has a width of 0.5-3.0 mm, or a length of 2000-2500 mm, preferably, the swab-body 402 has a width of 0.7-1.5 mm and a length of 2000-2500 cm.

[0171] The method comprises using a swab head is made of synthetic polyamide fibers and the swab-body is made of nylon fiber.

[0172] Step B) comprises introducing the sample inside a channel tube 207. More specifically, the sample is introduced by inserting a swab-head 401 in channel tube 207. The channel tube 207 is in turn placed inside the channel chamber 206.

[0173] Step C) comprises contacting a sample with a reagent composition.

[0174] In an embodiment, the reagent composition comprises a “pre-activated” reagent composition. Preferably, the pre-activated reagent composition comprises

[0175] • a dye suitable for protein detection, such as a triphenylmethane dye, preferably, a dye with affinity for proteins such as Coomasie Blue G, at a concentration of at most 2% w / w, e.g. between 0-2% w / w, preferably between 0.01-2% w / w, a diluent suitable for a dye comprising an alcohol group, preferably in a concentration of at most 98% w / w.

[0176] In another embodiment, the pre-activated reagent composition comprises

[0177] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninicacid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w;

[0178] • an oxidant capable of reacting with a protein such that allows the formation of a colored complex with the chelating agent,

[0179] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, at a concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0180] • water at a concentration of at most 80% w / w,

[0181] In another embodiment, the reagent composition comprises a first “non-activated” reagent composition, and a second “non-activated” reagent composition, wherein the first and second composition are separated. More specifically, the first non-activated reagent composition is contained in a reagent chamber 203, and the second nonactivated reagent composition is contained in a reading chamber 201.

[0182] In an embodiment, the first non-activated composition comprises

[0183] • a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w;

[0184] • water at a concentration of at most 96% w / w.

[0185] In an embodiment, the second non-activated composition comprises

[0186] • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninic acid, at a concentration of at most 4% w / w, e.g., between 0-4% w / w, preferably between 0.01-4% w / w.

[0187] • an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, orfunctional equivalents, ata concentration of at most 30% w / w, e.g., between 0-30% w / w, preferably between 0.01-30% w / w,

[0188] • water at a concentration of at most 80% w / w, Preferably, the contacting of the sample and the reagent composition is carried out inside a reading chamber 201. Preferably, the reading chamber 201 is made of a transparent material suitable for performing and measuring an optic signal-generating reaction inside thereof, for example, said transparent material comprises copolymers selected from PETG, PCTA, and PCTG.

[0189] The contacting of step C) is done by breaking the frangible membranes 209 and 209b using the cutting end 301 of the channel tube 207, and is performed inside the reading chamber 201. These membranes seal the reagent chamber 203 ensuring that the reagent composition is kept undisturbed and under preservation conditions until used. Thus, the step C) comprises the release of the reagent composition from the reagent chamber 203 into the reading chamber 203.

[0190] In an embodiment, the cutting end 301 of the channel tube 207 comprises a beveled edge. Preferably a beveled edge of a shape selected from a J-bevel, V-bevel, Compound bevel, Compound J-Bevel, or any of these with or without counterbore. In another embodiment, the beveled edge can cut at an angle of 20°-30°. The beveled edge can comprise a hollow bevel with a central groove.

[0191] The optic signal-generating reaction of Step C) comprises detecting changes in the absorbance of wavelengths of the mixture that can be registered by the electronic optical reader apparatus. Said optic signal-generating reaction comprises a colorimetric reaction or a fluorescence reaction.

[0192] Step C) comprises using an electronic optical reader to perform the optic signalgenerating reaction. Said electronic optical reader can be configured to receive a reading chamber 203, incubate the reading chamber 203 at a predetermined temperature and time necessary for performing an optic signal-generating reaction, and measure any wavelength absorbance changes that can take place inside the reading chamber 201.

[0193] In an embodiment, step C) comprises determining the absorbance value of the mixture in wavelengths of 300-1000 nm, preferably between 400-700 nm, more preferably between 550-580 nm.

[0194] In another embodiment, step C) comprises determining the absorbance value of the mixture in wavelengths between 550-580 nm for a colorimetric reaction, or wavelengths between 350-400 or between 430-520 nm for a fluorescence reaction. In an embodiment, step C) comprises determining the absorbance value of the mixture after an incubation of 40-70°C, preferably 60°C; during a period of 1-10 minutes, preferably 4 minutes.

[0195] Step D) further comprises determining the protein concentration in a sample by comparison with a standard calibration curve. The standard calibration curve can be performed using any protein suitable for such end, preferably, bovine serum albumin (BSA) is used as the standard protein. In an embodiment, the standard calibration curve can be used to determine the concentration of proteins between 0.5-60 pg.

[0196] EXAMPLES

[0197] In the following, the invention will be detailed in preferred embodiments described throughout the examples provided. These examples are to be considered only as exemplary embodiments of the invention and are not intended to limit the scope of the invention in any way.

[0198] EXAMPLE 1

[0199] 1.1. - Colorimetric Reaction

[0200] The term “colorimetric reactive solution” refers to a solution that generates a measurable absorbance response upon interaction with protein residues. This reactive solution is utilized in an assay method involving the introduction of varying concentrations of bovine serum albumin (BSA) into a defined volume of the reactive solution. The procedure includes preparing dilutions from a standard BSA solution with a concentration of 2 mg / mL, wherein a control sample devoid of protein is also incorporated. The mixtures undergo incubation for four minutes at a temperature of 60 °C, followed by immediate cooling to halt the reaction. The absorbance of the samples is measured using a spectrophotometer across the wavelength range of 300 to 1000 nm, with air serving as the baseline reference and a cuvette filled with water acting as the spectrum blank. A calibration curve (Fig. 5) is established by plotting the absorbance at 562 nm against the corresponding protein quantities in micrograms, thus creating a reliable standard for protein quantification within the reactive solution.

[0201] 1.2 - Electronic Optical Reader In another aspect, the present disclosure provides an electronic optical reader designed for detecting and quantifying protein residues on cannulated instruments. This system is able to carry out the colorimetric reaction in any embodiment of the device of the first aspect. The electronic optical reader is configured to: (i) receive the reading chamber, (ii) incubate the sample at a stable 60 °C to maintain consistent reaction kinetics, and (iii) measure or detect colorimetric changes in the colorimetric reactive solution, which indicates the presence of protein residues.

[0202] The automated reader includes, among other elements, a reading chamber, a calibrated light source for precise illumination, an RGB photodetector capable of detecting colorimetric change, and at least one microprocessor to control the components. Optionally, the apparatus may include software with specific algorithms for processing the readings and interpreting the results.

[0203] Example 3 - Protein residual detection

[0204] The ability of the method to detect protein residual was evaluated in various types of cannulated surgical instruments with different internal diameters, using swabs of different diameters (1.7 mm, 2.0 mm, 2.7 mm, and 3.0 mm). The results demonstrate that the method quantifies protein residual in all analyzed instruments, including cases where protein levels are zero, confirming high sensitivity, reproducibility, and reliability.

[0205] Protein recovery varies according to the instrument type and the relationship between the internal diameter and the swab diameter, but appropriate swab selection ensures consistent and reproducible measurements. Table 1 summarizes the ranges of quantified protein residuals, confirming that the method provides a precise and reliable assessment of cleaning and disinfection of cannulated surgical instruments.

[0206] Note: The “Protein residual (pg)” column represents quantified measurements of protein residual after cleaning and disinfection. All results were obtained using the swab method, demonstrating reliable quantification across instruments. < < <

[0207]

Claims

CLAIMS1. A device for monitoring the hygienic status of a cannulated instrument, wherein said device comprises:a reading chamber at one end of the device,a reagent chamber placed contiguously to the reading chamber and comprising two frangible membranes each located at opposite ends of the reagent chamber,a channel chamber placed contiguously to the reagent chamber such that the reagent chamber is placed between the reading chamber and the channel chamber, and wherein the channel chamber comprisesa channel tube having a cutting end for breaking the frangible membranes, placed within the channel chamber, anda piston-like cap at the other end of the device.

2. The device of claim 1, wherein the reagent chamber comprises a reagent composition having• a dye suitable for protein detection, such as a triphenylmethane dye, preferably, a dye with affinity for proteins such as Coomasie Blue G, at a concentration of at most 2% w / w,• a diluent suitable for a dye comprising an alcohol group, preferably in a concentration of at most 98% w / w.

3. The device of claim 1, wherein the reagent chamber comprises a first reagent composition having• a compound providing divalent metal ions in solution, such as Cu+2, at a concentration of at most 4% w / w,• water at a concentration of at most 96% w / w;and the reading chamber comprises a second reagent composition having • a chelating agent capable of forming colored complexes with metal ions, preferably bicinchoninic acid, at a concentration of at most 4% w / w,• an alkaline compound selected from the group comprising sodium carbonate, potassium carbonate, or functional equivalents, at a concentration of at most 30% w / w,• water at a concentration of at most 80% w / w.

4. The device of any one of claims 1-3, wherein the cutting end of the channel tube comprises a beveled edge.

5. An electronic optical reader apparatus for the detection and quantification of protein levels, comprising:a light emitter,a sensing unit placed in front of the light emitter,at least one reading well, comprising an orifice placed perpendicular to the light emitter and the sensing unit, wherein said orifice is located at the height at which both the sensor and emitter align with a reading chamber of a device of the first aspect, anda heating block in contact with the at least one reading well.

6. The apparatus of claim 5, wherein the light emitter comprises a source capable of emitting light at 542-582 nm.

7. The apparatus of claim 5, wherein the light emitter is at a distance between IQ- 20 mm from the reading well, and at a height of between 8.0-12.0 mm.

8. A method for monitoring the hygienic status of a cannulated instrument, said method comprising the steps of:a) taking a sample from the cannulated instrument,b) introducing the sample in a device according to any one of claims 1-4, c) contacting the sample with a reagent composition, thereby triggering an optic signal-generating reaction,d) determining the concentration of protein residues in the sample.

9. The method of claim 8, wherein the optic signal-generating reaction of step b) is performed at a temperature between 55-65°C, for a period of 1-10 minutes, and the absorbance is measured at 300-1000 nm.

10. The method of claim 9, wherein the optic signal generating reaction comprises a colorimetric reaction and the absorbance is measured at 550-580 nm.

11. The method of claim 9, wherein the optic signal-generating reaction comprises a fluorescence reaction and the absorbance is measured at between 300-400 nm or between 430-520 nm.

12. The method of any one of claims 8 to 11, wherein the determination of concentration of protein is carried out by using a standard calibration curve.

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