WO2018025046A2

WO2018025046A2 - Device

Info

Publication number: WO2018025046A2
Application number: PCT/GB2017/052291
Authority: WO
Inventors: Kodanda Ranganatha Nagaraja RAO; Carl Jorg Michael GRAZ
Original assignee: Nn Scientific Limited; Biophys Limited
Priority date: 2016-08-05
Filing date: 2017-08-04
Publication date: 2018-02-08
Also published as: WO2018025046A3

Abstract

The present invention provides an analyser suitable for use in the simultaneous diagnosis of one or more conditions using a sample obtained from a human or animal body, the analyser comprising a first chamber having a first entry port and a second chamber having a second entry port, wherein each chamber is pre-loaded with assay reagents. The analyser may additionally comprise a probe comprising: a first swab tip and a second swab tip; and an actuator configured to transition the probe between a sampling configuration and an analysis configuration, wherein in the sampling configuration the first swab tip and the second swab tip are located such that they form a single swab tip, and wherein in the analysis configuration the first swab tip and the second swab tip are located relatively further apart than in the sampling configuration such that they are operable as two separate swab tips. Kits and method relating to the same are also provided.

Description

Device

Field of Invention

The present invention relates to parallel diagnosis of pathological conditions and especially to simultaneous differential diagnosis of two or more different pathological conditions. Kits, apparatus and methods relating to the same are also described.

Background to the Invention

A number of pathological conditions are difficult to diagnose on the basis of clinical symptoms alone. For example, the symptoms of Candidiasis (yeast infection) and bacterial vaginosis (BV) both include itching and discharge.

Yeast infections are caused by an overgrowth of Candida microorganisms and are conventionally diagnosed based on clinical symptoms and/or potassium hydroxide (KOH) testing (where a sample is treated with KOH solution followed by microscopic examination to identify the presence Candida microorganisms) and or microscopy and more recently PCR analysis. However, both of these methods require a patient to be examined by a medical practitioner and access to laboratory facilities..

BV infections are caused by an overgrowth of anaerobic bacteria with a concomitant decrease in numbers of lactobacilli. The drop in lactobacilli numbers leads to an increase in pH, which is commonly used as a diagnostic marker for BV. Diagnosis is also based on clinical symptoms using the Amsel criteria where at least three of the four criteria are met: a characteristic vaginal discharge; vaginal pH of great than 5; a positive amine test with boiling KOH and a fishy odour (Whiff test); and the presence of "clue cells" on microscopy.

While yeast infections and BV can have very similar symptoms the causes of these conditions are very different, being fungal and bacterial infections, respectively, and different therapeutic agents are required for effective treatment. Currently, excepting the more expensive and lengthy PCR analysis, the more usually available clinic available diagnostic methods are only able to distinguish one or other of these conditions, which can make it difficult to make a correct diagnosis, especially if a patient is relying on over the counter (OTC) tests, rather than visiting their doctor. Treatments for a yeast infection do not effectively treat BV, and repeated BV infections can lead to increased susceptibility to HIV infection and infertility. Incorrect diagnosis of yeast infections can lead to overuse of antibiotics, which is well known to contribute to the growing problem of antibiotic resistance. There is therefore a need for a rapid simultaneous diagnosis of these conditions in order to provide effective and appropriate treatment both in clinic and home testing (over the counter, OTC) setting. Other conditions such as bacterial and viral chest infections or abdominal pain have similar needs.

The present invention therefore provides quick and economical methods for parallel diagnosis of pathological conditions and especially to simultaneous diagnosis of two or more different pathological conditions. Kits and apparatus relating to the same are also described.

Summary of the Invention

In a first aspect the present invention provides a probe suitable for collecting biological samples from a human or animal body, the probe comprising a handle connected to a first swab tip and a second swab tip, wherein the probe further comprises an actuator configured to transition the probe between a sampling configuration and an analysis configuration, wherein in the sampling configuration the first swab tip and the second swab tip are located such that they form a single swab tip, and wherein in the analysis configuration the first swab tip and the second swab tip are located relatively further apart than in the sampling configuration such that they are operable as two separate swab tips. The probe of the present invention therefore allows at least two samples to be collected from a single sampling step. This reduces costs and improves the efficiency of the diagnostic process. The single sampling step also has important advantages for patients in situations where sampling might be invasive, uncomfortable or embarrassing. Significantly, the probe of the present invention allows sufficient sample to be collected in the single sampling step to run at least two separate diagnostic tests.

The probe of the invention may be a single piece construction. For example, the probe may be extruded or 3D printed as a single piece construction. Alternatively, the probe may be vacuum cast or injection moulded. In embodiments of the invention the probe may be blow moulded. When provided as a single piece construction it is preferred that the swab tips are an integral part of the probe. In other words, the swab tips can be formed from the same material as the rest of the probe and no other parts or materials are added to them. The swab tips are preferably textured, e.g. through the use of ridges or dimples and do not comprise any added on elements such as brushes or buds.

In a second aspect the present invention provides a probe suitable for collecting biological samples from a human or animal body, the probe comprising a handle and a first receiving port for a first swab having a first swab tip and a second receiving port for a second swab having a second swab tip, wherein the probe further comprises an actuator configured to transition the probe between a sampling configuration and an analysis configuration, wherein in the sampling configuration the first swab tip and the second swab tip are located such that they form a single swab tip, and wherein in the analysis configuration the first swab tip and the second swab tip are located relatively further apart than in the sampling configuration such that they are operable as two separate swab tips.

Preferably the first and second receiving ports of the second aspect of the invention comprise grips for holding the first and second swabs. The grips allow swabs to be releasably connected to and held securely by the handle. In embodiments of the invention the grips comprise a female part into which the end of the swabs interlock as the corresponding male part. In more detail, the grips may compress a recess or dimple that enables the end of the swab to interlock with the handle.

The swabs used in the second embodiment of the invention may be conventionally known and commercially available swabs, such as cotton wool swabs. Such swabs are relatively inexpensive and widely available. Suitable swabs can be inserted into the handle for the sampling steps. The swabs may be detached from the handle for the analysis step or may be left in place. Prior to taking a further sample from the same patient or taking a sample from a different patient the used swabs can be removed from the handle and new swabs can be inserted. This avoids the need to provide a new handle for each sampling step.

The probe including the handle and optionally, the swabs, is preferably formed from a biologically inert material that is amenable to conventional types of sterilisation, such as autoclaving or gamma irradiation. The probe may therefore be formed from or comprise plasticised paper, plastic or a polymer. Suitable materials may include e.g. polyurethane, polyethylene, polycarbonate and/or polyvinyl chloride (PVC). Preferably the material has a high degree of flexibility, which aids the transitions between the sampling and analysis configurations. The material is preferably shatter proof and resistant to breakage. In preferred embodiments of the invention the material is hydrophilic. Optionally the probe material may be biodegradable.

The swab tips may be formed from a solid, open or closed cell foam or may be formed from cotton or viscose. In embodiments of the invention the swab tips may formed from a biologically inert solid, such as a plastic or polymer. Suitable materials may include e.g. polyurethane, polyethylene, polycarbonate and/or polyvinyl chloride (PVC). In embodiments of the invention the swab tips may be textured, e.g. through the use of ridges or dimples, to aid in retention of the biological sample. The surface texture of the swab tip can provide particular advantages in allowing a sufficient volume of sample to be held by the swab to react with an analyte when the sample is tested.

In embodiments of the probe the first swab tip and second swab tip may be resiliently biased toward the sampling configuration. Alternatively, the first swab tip and second swab tip may be resiliently biased toward the analysis configuration. Suitable mechanisms for achieving the resilient bias will be familiar to the skilled person. For example the structure of the handle of the probe may bias the swab tips toward the sampling or analysis configurations. The resilient bias may be provided by a slight bowing to the handle, preferably biasing the swab tips toward the sampling configuration. Additionally or alternatively, the resilient bias may be provided by a spring. Preferably the spring is an integral part of the handle of the probe and is formed from the same material as the rest of the handle. When the swab tips are resiliently biased toward the analysis configuration the probe preferably comprises a locking means that locks the swab tips in the sampling configuration until the probe is ready for analysis.

The actuator may transition the probe from the sampling configuration to the analysis configuration or from the analysis configuration to the sampling configuration or both. When the swab tips are resiliently biased toward the sampling configuration the actuator preferably transitions the probe from the sampling configuration to the analysis configuration. When the swab tips are resiliently biased toward the analysis configuration the actuator preferably transitions the probe from the analysis configuration to the sampling configuration.

The actuator may comprise a lever configured to pivot about a fulcrum to transition the probe between the sampling configuration and the analysis configuration. Additionally or alternatively the actuator may comprise a handle that is compressed to transition the probe between the sampling configuration and the analysis configuration. The actuator may comprise a spring, such as a V-spring. In embodiments of the invention the actuator may form part of the handle of the probe and squeezing the actuator urges the swab tips from the sampling configuration to the analysis configuration. Preferably the actuator is an integral part of the handle of the probe and is formed from the same material as the rest of the probe. The actuator may comprise a living hinge.

As mentioned above, when in the sampling configuration the swab tips form a single swab tip. In other words, in the sampling configuration the swab tips are substantially in contact with each other, for example as pressed together by the resilient bias toward the sampling configuration. In the sampling configuration preferably at least some of the outer surface of the first swab tip is in contact with at least some of the outer surface of the second swab tip. For example, at least 20% or at least 30% or at least 40% of the outer surface area of the first swab tip may be in contact with the second swab tip.

Preferably when in the analysis configuration the swab tips are separated to allow the sample collected on each tip to be analysed separately. The swab tips are therefore positioned away from each other in the analysis configuration and are preferably not in contact with each other. The optimum configuration for the analysis configuration will be determined by the way in which the swab tips are analysed. For example, the analysis configuration may be arranged such that the first and second swab tips align with the first and second entry ports of the analyser described below. In preferred embodiments of the invention the first and second swab tips may be substantially parallel when in the analysis configuration.

The probe may comprise a chemical or biological agent for detecting a substance in the biological sample and a display for indicating the presence or absence of said substance in the sample. For example, the probe may comprise one or more of a pH indicator, a pH dependent or pH independent redox indicator, a chromophore or fluorophore, such as fluorescamine, that changes colour in the presence of a desired substance in the biological sample. The display may comprise a window allowing the colour change to be viewed by the user and/or may be in the form of an electronic display. In preferred embodiments of the invention the substance in the biological sample is a pathogen such as a bacteria, virus or fungus.

In a third aspect the present invention provides an analyser suitable for use in the diagnosis of one or more conditions using a sample obtained from a human or animal body, the analyser comprising a first chamber having a first entry port and a second chamber having a second entry port, wherein each chamber is pre-loaded with assay reagents. In preferred embodiments of the invention the first and second chambers are loaded with different assay reagents. The analyser is therefore able to simultaneously diagnose the presence or absence of two different conditions.

Preferably the first and second chambers each comprise an upper compartment and a lower compartment, separated by a breakable seal. The analyser of the present invention is therefore a reactor, containing at least two separated components, which are combined to form an active species, which then reacts with a sample.

In embodiments of the invention the upper and lower compartments may each comprise a separate seal. For example, breakable seals may be placed at the lower end of the upper compartment and/or at the upper end of the lower compartment. The upper end of the upper compartment may also be covered by a breakable or peelable seal. Preferably reagents in the upper and lower compartments are physically and chemically separate until such time as the seal is broken. Consequently, the seal is preferably gas and liquid impermeable and can comprise or be made of a non-woven material such as a plastic or foil. The upper and lower compartments of each chamber provide particular advantages in that reagents in the upper and lower compartments can be kept separate until required for testing. This is beneficial because when mixed the reagents form reactive species, generally having a short half-life, which may be as little as 10 minutes. Keeping the reagents separate until required also avoids false positives such as those that may be caused by oxygen reacting with reagents to cause a colour change.

In embodiments of the invention the seal or seals can be broken by squeezing the outer walls of the analyser. Alternatively, the seal or seals may be broken by twisting or pressing upper and lower portions of the analyser toward each other, such movement causing the seal(s) to be pierced or otherwise broken. In embodiments of the invention the seal(s) may be broken by the insertion of a probe as described herein. Once the seal or seals have been broken, assay reagents in an upper compartment may drop or flow into assay reagents of the corresponding lower compartment, causing the formation of reactive species. Alternatively, the biological sample may be dispersed in liquid assay reagents of an upper chamber prior to the seal being broken. The liquid assay reagents comprising the dispersed sample will then react concomitantly with the assay reagents in the corresponding lower compartment once the seal has been broken.

In embodiments of the invention the upper and lower portions of the analyser can be releasably coupled, e.g. using threads or snapfit. Alternatively, the upper and lower portions of the analyser may be non-releasably coupled. In embodiments of the invention once the upper and lower portions of the analyser are twisted or pressed together they interlock, e.g. though a snap fit, which acts to prevent the upper and lower portions of the analyser from being separated. This can reduce the risk of the user or environment being exposed to the reagents or any potentially infectious sample contained within the analyser.

The analyser can comprise or be made of plastics material, especially polypropylene or polyethylene, low or high density, polyvinylchloride or other polymer used in manufacture of containers or in the medical industry, e.g. polyethylene terephthalate, polyolefin or polycarbonate. The analyser may be manufactured by extrusion, vacuum casting, injection moulding or 3D printing.

In embodiments of the invention at least a portion of the lower portion of the analyser is substantially transparent so as to allow a colour change or other signal to be visible to the user or to an optical reader. Preferably at least a portion of the lower compartments of the first and second chambers is substantially transparent. In embodiments of the invention the entire lower portion of the analyser may be transparent.

In embodiments of the invention the analyser may comprise an outer wrap substantially covering the first and second chambers, the wrap preferably being formed from a flexible material such as card, foil or plastic. The wrap may comprise a cut out or transparent window allowing at least a portion of the lower compartments of the first and second chambers to be visible. The wrap may be printed with a colour chart or other details relating to the analyser, such as instructions for use. Preferably the assay reagents react to indicate the presence or absence of a substance in the biological sample. For example, the assay reagents may react in the presence of a pathogenic substance such as a bacteria, virus, protozoa or fungus. Additionally or alternatively, the assay reagents may react in the presence of a biomarker. The biomarker may be a marker for a pathological substance (such as a bacteria, virus, protozoa or fungus), or it may be a biomarker for a disease condition. The reaction may take the form of a colour change, which may be visible through a transparent window in a portion of the analyser. Additionally or alternatively, the presence of the substance may be indicated by an electronic signal such as may be provided by a light or an electronic display.

In embodiments of the invention the analyser may be inserted into an optical reader to detect a colour change in the assay reagents.

The presence or absence of the substance in the biological sample is preferably indicated within about 10 minutes or less, more preferably within about 5 minutes or less. In other words, once the biological sample has been introduced to the analyser any colour change or other signal of positive diagnosis will occur within about 5 minutes or less. The analyser is typically incubated at room temperature during this period.

The type of assay reagents are not particularly limited. However, it is preferred that the assay reagents have a strong and obvious colour change in the presence of a target substance, the colour change being visible under normal visible light. For example, the assay reagents may present as a pale green in the absence of the substance, changing to a bright blue in the presence of the substance, or pale yellow in the absence of a substance, changing to pink in the presence of the substance. This helps to ensure quick and simple detection of the substance enabling a diagnosis to be made without the need for complex equipment or specialist training. Colour charts may be provided on the analyser or wrap to assist the user in making a diagnosis. In embodiments of the invention the upper compartments may comprise solid or liquid assay reagents and the lower compartments comprise solid or liquid assay reagents. Solid assay reagents may comprise reagents in the form of dry powders or granules. In use, when the seal between the upper and lower compartments is broken the assay reagents of the upper compartment drop or flow into the lower compartment, where they combine with the assay reagents of the lower compartment to form reactive species. In preferred embodiments of the invention the upper compartments comprise liquid assay reagents and the lower compartments comprise solid assay reagents.

The total volume of reagent in the compartments of the analyser may be about 5ml or less, about 3ml or less, preferably about 2ml or less. In preferred embodiments of the invention the total volume of reagent in the compartments is about 1ml or less.

The assay reagents may include one or more of a pH dependent or pH independent redox indictor, a pH indicator, a chromophore or a fluorophore, such as fluorescamine, that changes colour in the presence of a desired substance in the biological sample. Alternatively, the assay reagents may comprises antigens or antibodies covalently linked to enzymes to provide an enzyme-linked immunosorbent assay (ELISA) that can produce a visible signal in the presence of a desired substance (such an antigen or antibody) in the biological sample.

The assay reagents of the upper compartments may be selected from one or more of buffer, dimethyl formamide, dimethyl sulphoxide and water.

The assay reagents of the lower compartment may be selected from one or more of the following combinations: urea hydrogen peroxide/3,5 dihydroxy phenylalanine; Indigo disulphonic acid/potassium; carbonate; Indigo tetrasulphonic acid/potassium carbonate; hydrogen peroxide/o-phenylenediamine; Safranin T/potassium carbonate; Phenol Red in glycine- or ammonium formate-potassium chloride; or Alizarin Red S in potassium dihydrogen phosphate-potassium chloride. The analyser is preferably suitable for use with a probe according to the first or second aspect of the invention. In preferred embodiments of the invention once the probe has been inserted into the analyser the components interlock to form a seal that prevents the sample or assay reagents from exiting the analyser. Said interlocking may be aided or provided by the resilient bias of the swab tips. For example, if the swab tips are resiliently biased toward the sampling configuration they will try to revert to this configuration once the actuator has been released, thereby causing the swab tips to gip or squeeze the insides of the chambers of the analyser.

In a fourth aspect the present invention provides a kit comprising a probe and an analyser as described herein, wherein the analyser is complementary in shape to the analysis configuration of the probe such that the first swab tip can be inserted via the first entry port into the first chamber simultaneously to the second swab being inserted via the second entry port into the second chamber. The kit therefore allows a single sample to be simultaneously tested in two separate sets of reagents, which may be the same or different. In this way the single sample can be simultaneously tested, for example to diagnose two different conditions, or to test for a single condition in two different ways. The arrangement of entry ports and chambers in the analyser prevents intermixing of reagents between the chambers.

The kit may further comprise instructions for use and is preferably in an over the counter form. The kit may therefore be purchased without prescription. Preferably the kit is suitable for use both in the home and in a clinical setting. Typically, the kit is portable.

The kit may further comprise one or more active therapeutic agents for the treatment of conditions diagnosed by the kit. For example, the active therapeutic agents may be selected from an antibacterial agent, an antiviral agent and an antifungal agent.

In a fifth aspect the present invention a method of parallel diagnosis, the method comprising the steps of: i) providing a biological sample simultaneously to the first and entry ports of an analyser as described herein, and ii) allowing the assay reagents to react with the biological sample in two parallel assays to provide two parallel output signals. The method therefore allows a single sample to be simultaneously tested in two separate sets of reagents, which may be the same or different. In this way the single sample can be simultaneously tested to diagnose two different conditions, or to test for a single condition in two different ways. For example a urine sample may be simultaneously tested for a marker of appendicitis and a marker of inflammation, such as the presence of leukocytes, thereby providing a test for appendicitis having improved specificity.

The biological sample may be provided by a pipette or dropper, or may be provided by a probe as described herein. When the biological sample is provided by a probe of the present invention the method preferably comprises providing the biological sample simultaneously to the first and second swab tips of the probe, manipulating the actuator to transition the probe from the sampling configuration to the analysis configuration, and inserting the probe into the analyser such that the first swab tip is inserted via the first entry port into the first chamber simultaneously to the second swab tip being inserted via the second entry port into the second chamber. The method may be carried out using a kit as described above.

The biological sample may be any sample obtained from a mammal including humans. In embodiments of the invention the sample may be a sample of bodily fluid, such as urine, mucus or faecal matter. The sample may be obtained from a bodily orifice, such as the nose mouth, throat, ears, vagina, urethra or anus.

Preferably the method provides for simultaneous diagnosis of two different conditions, such as two different infections. The conditions may be selected from a bacterial infection, a viral infection and/or a fungal infection. For example, the conditions may be a bacterial infection and a viral infection, a bacterial infection and a fungal infection or a fungal infection and a viral infection. Alternatively, the conditions may be two different bacterial infections, two different fungal infections or two different viral infections. Preferably, the conditions are a bacterial infection and a fungal infection, such as a Candida infection and a bacterial vaginosis infection. In embodiments of the invention the infection may be a bacterial infection associated with a mammalian lung, or it may be a Candida infection in a male. The device may also be used for diagnosis of a urinary tract infections or distinguishing between appendicitis and pelvic inflammatory disease.

The parallel output signals may be in the form of a colour change or an electronic reading as described above.

In embodiments of the invention the method may further comprise administering a suitable treatment for the diagnosed conditions.

Brief Description of the Drawings

Figures 1 to 4 show analysers of the present invention as described herein.

Figures 5 to 11 show probes of the present invention as described herein.

Figure 12 shows the results of Candida and bacterial vaginosis tests as carried out in Example 14. Panel A shows the presence (dark purple/black) and absence (pale brown) of Candida. Panel B shows the presence (reddish pink) or absence (pale yellow) of polyamines such as putrascine and cadaverine, which are markers of infection due to bacterial vaginosis. Panel C shows the absorbance of the positive bacterial vaginosis test at a wavelength of λ515ηιη.

Figure 13 shows the results of Candida and bacterial vaginosis tests as carried out in Example 16. Panel A shows the presence (dark purple/black) and absence (pale brown) of Candida. Panel B shows the presence (reddish pink) or absence (pale yellow) of polyamines such as putrascine and cadaverine, which are markers of infection due to bacterial vaginosis. Panel C shows the absorbance of the positive bacterial vaginosis test at a wavelength of λ525ηιη. Figure 14 shows the results of sensitivity tests comparing Phenol Red with Indigo Carmine. Panel A shows a 25 to 500 μg range for Phenol Red. Panel B shows a 50 to 500 μg range for Indigo Carmine.

Figure 15 shows the presence (dark purple/black) or absence (pale brown) of Candida in panels A, C and E. Panel B shows the presence (blue) or absence (pale green) of bacterial vaginosis using Indigo Carmine. Panel D shows the presence (red) or absence (pale yellow) or bacterial vaginosis using Phenol Red. Panel F shows the presence (red) or absence (pale yellow) or bacterial vaginosis using Alizarin Red S. Panel G shows the absorbance of a positive test for BV at a wavelength of 525 nm.

Description

Referring now to Figure 1, there is shown a view of an analyser of the present invention. The analyser comprises a circular housing 1 having an upper lid 2 with two circular entry ports 3A,3B along the length thereof, a lower end piece 4 comprising two chambers, each divided into a top compartment 5A and a bottom compartment 5B. In the illustrated embodiment, the entry ports 3A,3B comprise two semi-circular cut outs that extend towards the lower end 4. The entry ports 3A,3B are adapted to provide access to a probe or other means for the introduction of test analytes. Prior to use of the analyser the entry ports 3 A,3B can be sealed by a removable lid 8, such as a peelable foil lid. The analyser allows reagents in the upper and lower compartments of each chamber to come in contact with each other through an opening (not shown) created by squeezing the outer part of the analyser together. In a preferred embodiment, the housing of the analyser is composed of plastic or other suitable materials such as polypropylene, polyethylene, polyvinyl chloride or polycarbonate.

In the example provided, the upper and lower compartments of each chamber are separated by a breakable seal 6, such as a thin strip of non-porous material that can be broken open by squeezing the outer part of the device. The analyser may comprise a wrap 9, such as a card, foil or plastic wrap, covering the two chambers. Preferably the wrap comprises a test window (not shown). The test window may comprise a rectangular cut out (not shown) that allows the user to view a portion of the test colour change, which is disposed within the interior of the housing. The analyser may be adapted to display a colour chart 7 for showing a positive result and for comparisons.

The housing 1 comprises compartments providing a hollow interior for holding reagents therein. The reagents in each of the compartments may be in a form of a solid or liquid or a combination of solid and liquid. The exterior of the analyser may comprise a protrusion (not shown) that can be pressed towards the interior of the housing. When pressed, the breakable seal 6 breaks to allow the reagents from the upper compartment 5A to combine with the lower compartment 5B and remain to form an active substrate.

Referring now to Figure 2, there is shown an embodiment of the analyser of the present invention which comprises a circular housing having an upper lid 1 1 with two circular entry ports 12A,12B along the length thereof and a lower end piece 13 comprising two chambers, each divided into a top compartment and a bottom compartment. In the illustrated embodiment, the entry ports 12A,12B comprise two circular cut-outs that extend towards the lower end of the analyser but are separated from the lower end piece 13 by a seal 18. The entry ports 12A,12B are adapted to provide access for a swab or for the introduction of test analytes. The analyser allows reagents in the upper and lower compartments to come into contact with each other through an opening (not shown) created by squeezing the outer part of the analyser together to break a seal separating the upper and lower compartments. In a preferred embodiment, the housing of the analyser is composed of plastic or other suitable materials.

In the example provided, the upper and lower compartments are separated by a breakable seal in the form of a thin strip of non-porous material (not shown) that can be broken open by a twisting the upper lid of the analyser while pushing the upper and lower parts toward each other. In more detail, the upper lid 11 and lower end piece 13 of the analyser are separated by an internal seal 18 and an outer security seal 15 comprising a pull tab 14, which is used to remove the security seal. The upper lid 11 is then twisted such that helical thread 17 in the top part of the device engages with a mating thread 20 in the lower part of the analyser, thereby causing spikes 19 to pierce internal seal 18. This operation breaks open the internal seal between the upper lid 1 1 and lower end piece 13, as well as breaking the seal between the upper and lower compartments, thus allowing the reagent housed in the upper compartment to travel towards the lower compartment. The two reagents coming into contact generates reactive species. Introduction of test analytes into the reactive species will trigger a cascade of reactions to give coloured products.

The housing of the analyser also comprises a test window 16. In the illustrated embodiment, the test window comprises a rectangular cut out in wrap 21 that allows the user to view a portion of the test colour change, which is disposed within the interior of the housing. The device is adapted to display a colour chart (not shown) for showing a positive result and for comparisons.

Referring now to Figure 3, an embodiment of the analyser of the present invention comprises a rectangular housing 25 having an upper compartment 26 including housing for aqueous reagents 27 A, 27B. Bottom compartments 28 contain solid reagents. Reagents in the top and bottom compartments are allowed to come in contact with each other through an opening created by removing the security strip using pull tab 24 and pressing the upper compartment 26 and bottom compartments 28 together. In a preferred embodiment, the housing 25 is composed of plastic or other suitable materials. The analyser may additionally comprise a gasket 34, which prevents leakage from the device.

The top and bottom compartments may be separated by a breakable seal 29, such as may be provided by a thin strip of non-porous material that can be broken open by squeezing the outer part of the analyser. By pressing the upper lid 26 and lower end piece 28 of the device together, sheer 33 and spikes 31 pierce seal 29 and the reagent located in the top compartment migrates to the bottom compartment, thus allowing a reactive species to form. Introduction of a test sample will trigger a sequence of reactions culminating in a discrete colour change of the reagents in the bottom compartment. This colour change can be visualised through a test window 30. As shown in Figure 3, the test window 30 may comprise a rectangular cut out that allows the user to view a portion of the reactive species. The analyser may be adapted to display a colour chart (not shown) for showing a positive result and for comparisons. The colour chart may be printed on outer wrap 32.

Figures 4A and 4B show an embodiment of the analyser of the present invention having an upper lid 60, with two entry ports 61A, 61B. Chambers 62A, 62B are divided into upper compartments 63A, 63B in the lid 60 and lower compartments 64A, 64B in lower end piece 67. The lower ends of the upper compartments 63 A, 63B are sealed by breakable seals 65 A, 65B, while the upper ends of lower compartments 64A, 64B are sealed by breakable seals 66A, 66B. Pressing upper lid 60 onto lower end piece 67 causes the bottom compartments 64A, 64B of each chamber to pierce the breakable seals, allowing reagents in the upper and lower compartments to mix.

The analyser may comprise a locking mechanism (not shown), such as a pin, which prevents the upper lid and lower end piece from being pressed together until the user is ready to break the seals, i.e., the user may be required to remove the pin before the upper lid 60 and lower end piece 67 can be pressed together. In embodiments of the invention entry ports 61A and 61B may comprise additional seals (not shown) that can be removed prior to the introduction of test analytes. The seals may be breakable seals that can be pierced by a swab or they may be peelable seals.

In all of the four analysers illustrated above, the reagent located in the top section of a compartment migrates towards the bottom section of that compartment to give a reactive species or a reactive substrate. Introduction of a test sample via a swab or other means allows a series of reactions to occur that culminates in the generation of colour species or substances that can be easily read out.

Figures 5 to 11 illustrate probes of the present invention. As shown in Figures 5 to 1 1 a probe comprises a handle 35 connected to first and second swabs 36A, 36B, each swab having a swab tip 37 for retaining a sample. In a sampling configuration swabs 36A,36B are arranged such that swab tips 37 form a single swab tip. In an analysis configuration swab tips 37 are separated such that each tip aligns easily with entry ports 38A,38B in an analyser. Swab tips 37 may be in any form suitable for retaining a biological sample, for example, the swab tips may be formed from a solid open or closed cell foam or may be formed from cotton wool. In embodiments of the invention the swab tips may be textured, e.g. through the use of ridges or dimples, to aid in retention of the biological sample.

As shown in Figure 7, swab tips 37 may be joined together when the probe is in the sampling configuration, e.g. via perforations 40 between the two swab tips. The probe may comprise a living hinge 51, such that pushing the ends of swabs 36A, 36B toward each other causes perforations 40 to split, separating the swap tips 37 and transitioning the probe into the analysis configuration.

As shown in Figures 8 and 10, swabs 36A,36B comprising swab tips 37 may be removable from handle 35. In this embodiment of the invention swabs 36A,36B are situated within first and second receiving ports 41 A, 4 IB.

As shown in Figures 8 to 11, swabs 36A,36B may be reciprocally biased toward each other such that the probe is maintained in the sampling configuration. Application of pressure to actuator 42 transitions the probe to the analysis configuration by moving swab tips 37 away from each other. As shown in Figure 11, at least one of the swabs 36A may be shaped to allow the tip of the adjacent swab 36B to sit as close as possible to the tip of the first swab when the probe is in the sampling configuration.

Examples

Examples 1-5

The top compartments (26) of the analyser shown in Figure 3 contain liquids, as shown in Table 1. The bottom compartments (28) of the analyser contain a dry composite mixture as shown in Table 1. Active reagents are prepared by peeling off the security strip using the pull tab 24 and pressing the top and bottom parts together to allow the liquids in the top compartments to flow through into the bottom compartments. After allowing the reagents to mix together by gently shaking the analyser, the swabs with the analytes are introduced into the bottom compartments through the entry ports. The contents of the analyser are allowed to stand at room temperature and colour changes are compared with the chart. Colour changi indicative of the presence or absence of infection are as shown in Table 1.

Table 1: Reagents and indications used in examples 1-5.

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Compartments Compartments (dry Indication (liquid) composite)

A distinct blue colour is indicative of the pH 10 buffer, Urea hydrogen presence of polyamines like putrascine and peroxide/3,5 cadaverine. A green or pale yellow colour in dihydroxy phenyl this compartment is indicative of the alanine absence of infection due to bacterial

Example vaginosis.

1

A distinct deep dark purple or black colour dimethyl Indigo disulphonic

in the adjacent compartment is indicative of formamide acid/potassium

the presence of Candida while a pale brown solution, water carbonate

colour signifies absence of Candida.

A distinct blue colour is indicative of the pH 10 buffer, Urea hydrogen

presence of polyamines like putrascine and peroxide/3,5

cadaverine. A green or pale yellow colour in dihydroxy phenyl

this compartment is indicative of the alanine

absence of infection due to bacterial

Example

vaginosis.

2

A distinct deep dark purple or black colour dimethyl Indigo disulphonic

in the adjacent compartment is indicative of sulphoxide acid/potassium

the presence of Candida while a pale brown solution, water carbonate

colour signifies absence of Candida.

pH 10 buffer, Urea hydrogen A distinct blue colour is indicative of the peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A green or pale yellow colour in alanine this compartment is indicative of the

Example absence of infection due to bacterial

3 vaginosis.

A distinct deep dark purple or black colour dimethyl Indigo tetrasulphonic

in the adjacent compartment is indicative of formamide acid/potassium

the presence of Candida while a pale brown solution, water carbonate

colour signifies absence of Candida.

A distinct blue colour is indicative of the pH 10 buffer, Urea hydrogen

presence of polyamines like putrascine and peroxide/o- cadaverine. A green or pale yellow colour in phylenediamine

this compartment is indicative of the absence of infection due to bacterial

Example vaginosis.

4

A distinct deep dark purple or black colour dimethyl Indigo disulphonic

in the adjacent compartment is indicative of sulphoxide acid/potassium

the presence of Candida while a pale brown solution, water carbonate

colour signifies absence of Candida. pH 10 buffer, Urea hydrogen A distinct reddish violet is indicative of the peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A pale yellow colour in this alanine compartment is indicative of the absence of

Example infection due to bacterial vaginosis.

5

A distinct deep dark purple or black colour dimethyl

Sfranin T/potassium in the adjacent compartment is indicative of sulphoxide

carbonate the presence of Candida while a pale brown solution, water

colour signifies absence of Candida.

absence of infection due to bacterial

Example vaginosis.

6

Indigo Trisulphonic A distinct deep dark purple or black colour dimethyl

acid potassium in the adjacent compartment is indicative of formamide

salt/potassium the presence of Candida while a pale brown solution, water

carbonate colour signifies absence of Candida.

Examples 7-12

The top compartments (26) of the analyser shown in Figure 3, contain liquids as shown in Table 2. The bottom compartments (28) of the analyser contain a composite mixture of components as shown in Table 2. A probe as shown in Figure 6 containing the test samples is introduced in the upper compartments containing the solutions. After allowing the samples to disperse into the solutions for 2 minutes, the solution along with the swab tips are allowed to pass through into the bottom compartment by pressing the top and bottom compartments together to break the seal 29. The liquid portion along with the swab tips is thoroughly mixed with the solid reagent by gently shaking the analyser. The contents of the analyser are allowed to stand at room temperature and the colour change compared with the chart. Colour changes indicative of the presence or absence of infection are as shown in Table 2.

Table 2: Reagents and indications used in examples 7-12.

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Compartments Compartments (dry Indication

(liquid) composite) Example buffer of pH 10 urea hydrogen A distinct blue colour is indicative of the

7 peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A green or pale yellow colour alanine in this compartment is indicative of the absence of infection due to bacterial vaginosis. a solution of Indigo disulphonic A distinct deep dark purple or black colour Dimethyl acid/potassium in the adjacent compartment is indicative of formamide and carbonate the presence of Candida while a pale brown water colour signifies absence of Candida.

Example a buffer of pH 10 urea hydrogen A distinct blue colour is indicative of the 8 peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A green or pale yellow colour alanine in this compartment is indicative of the absence of infection due to bacterial vaginosis. a solution of Indigo disulphonic A distinct deep dark purple or black colour Dimethyl acid/potassium in the adjacent compartment is indicative of sulphoxide and carbonate the presence of Candida while a pale brown water colour signifies absence of Candida.

Example a buffer of pH 10 urea hydrogen A distinct blue colour is indicative of the 9 peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A green or pale yellow colour alanine in this compartment is indicative of the absence of infection due to bacterial vaginosis. a solution of Indigo tetrasulphonic A distinct deep dark purple or black colour Dimethyl acid/potassium in the adjacent compartment is indicative of formamide and carbonate the presence of Candida while a pale brown water colour signifies absence of Candida.

Example buffer of pH 10 urea hydrogen A distinct blue colour is indicative of the 10 peroxide/o- presence of polyamines like putrascine and phylenediamine cadaverine. A green or pale yellow colour in this compartment is indicative of the absence of infection due to bacterial vaginosis.

a solution of Indigo disulphonic A distinct deep dark purple or black colour Dimethyl acid/potassium in the adjacent compartment is indicative of sulphoxide and carbonate the presence of Candida while a pale brown water colour signifies absence of Candida. Example buffer of pH 10 urea hydrogen A distinct reddish violet is indicative of the 11 peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A pale yellow colour in this alanine compartment is indicative of the absence of infection due to bacterial vaginosis.

a solution of Sfranin T/potassium A distinct deep dark purple or black colour Dimethyl carbonate in the adjacent compartment is indicative of sulphoxide and the presence of Candida while a pale brown water colour signifies absence of Candida.

Example buffer of pH 10 urea hydrogen A distinct blue colour is indicative of the 12 peroxide/3,5 presence of polyamines like putrascine and dihydroxy phenyl cadaverine. A green or pale yellow colour alanine in this compartment is indicative of the absence of infection due to bacterial vaginosis. a solution of Indigo trisulphonic A distinct deep dark purple or black colour Dimethyl acidpotassium in the adjacent compartment is indicative of formamide and salt/potassium the presence of Candida while a pale brown water carbonate colour signifies absence of Candida.

Examples 13-16

The top compartments (26) of the analyser shown in Figure 3, contain reagent as shown in Table 3. The bottom compartments (28) of the analyser contain a composite mixture of reagents as shown in Table 3. The contents of the top compartments are allowed to pass into the bottom compartments by tearing out the security strip using pull tab 24 and pressing the device together to form a tight locking grip. This operation allows the liquid from the top compartment to flow into the bottom compartment. The liquid portion is allowed to mix with the solid reagent thoroughly by gentle shaking of the analyser for about 2 minutes. Upon this time period Glycine- potassium chloride dissolves to give a pale orange yellow coloured solution, while the solution in the second set of chambers where UHP-DOPA are allowed to mix with buffer, the colour of the active ingredient slightly pale brown. At this point a probe as shown in Figure 6 is introduced all the way into the bottom compartment. This operation locks the analyser securely. The contents of the analyser are allowed to stand at room temperature for 5 minutes with occasional shaking and resultant colour changes are compared with the chart. Colour changes indicative of the presence or absence of infection are as shown in Table 3.

Table 3: Reagents and indications used in examples 13-16.

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Indication

Compartments Compartments

Example buffer of pH 10 urea hydrogen A distinct reddish pink colour is indicative 13 (1.5 mL) peroxide/3,5 of the presence of polyamines like dihydroxy phenyl putrascine and cadaverine. A yellow alanine (35 mg) coloured solution in this compartment is indicative of the absence of infection due to bacterial vaginosis. solution of Phenol Red (18 μg in A distinct deep dark purple or black colour Dimethyl Gly cine-potassium in the adjacent compartment is indicative of sulphoxide and chloride (2: 1; 50 mg) the presence of Candida while a pale brown water (1:2;0.75 colour signifies absence of Candida.

mL)

Example buffer of pH 10 urea hydrogen A distinct reddish pink colour is indicative 14 (1.5 mL) peroxide/3,5 of the presence of polyamines like dihydroxy phenyl putrascine and cadaverine. A yellow alanine (35 mg) coloured solution in this compartment is indicative of the absence of infection due to bacterial vaginosis.

The reddish pink coloured solution thus formed has a distinct absorbance at λ₅ΐ5 nm in the ultraviolet region of the spectra (see Figure 12). solution of and Phenol Red (18 A distinct deep dark purple or black colour Dimethyl μg in ammonium in the adjacent compartment is indicative of sulphoxide and formate-potassium the presence of Candida while a pale brown water (1:2;0.75 chloride (2: 1; 50 mg) colour signifies absence of Candida.

mL)

Example buffer of pH 10 urea hydrogen A distinct reddish pink colour is indicative 15 (1.5 mL) peroxide/3,5 of the presence of polyamines like dihydroxy phenyl putrascine and cadaverine. A yellow alanine (35 mg) coloured solution in this compartment is indicative of the absence of infection due to bacterial vaginosis.

solution of Alizarin Red S (64 μg A distinct deep dark purple or black colour Dimethyl in potassium in the adjacent compartment is indicative of sulphoxide and dihydrogen the presence of Candida while a pale brown water (1:2;0.75 phosphate-potassium colour signifies absence of Candida.

mL) chloride (2: 1; 50 mg) Example buffer of pH 10 urea hydrogen A distinct reddish pink colour is indicative 16 (1.5 mL) peroxide/3,5 of the presence of polyamines like dihydroxy phenyl putrascine and cadaverine. A yellow alanine (35 mg) coloured solution in this compartment is indicative of the absence of infection due to bacterial vaginosis.

The pink coloured solution thus formed has a distinct absorbance at λ₅25 nm in the ultraviolet region of the spectra (see Figure 13). solution of Alizarin Red S (64 μg A distinct deep dark purple or black colour Dimethyl in potassium in the adjacent compartment is indicative of sulphoxide and dihydrogen the presence of Candida while a pale brown water (1 :2;0.75 phosphate-potassium colour signifies absence of Candida.

mL) chloride (2: 1; 50 mg)

Example 17

Standard solutions of either putrascine or Cadaverine were prepared in aqueous Dimethyl sulphoxide to give a concentration of 8.6 mg/mL. Aliquots of this solution (25,50,75,150, 200) were added to activated species prepared by mixing solid reagent with aqueous dimethylsulphoxide. After allowing the reaction to proceed for five minutes at RT the colour formation was noted and confirmed by absorption in the visible wavelength of a scanning ultraviolet visible spectrophotometer. From this study the lower limit of detection for this test was 21 micrograms of amine, (see Figure 14.)

Claims

1. An analyser suitable for use in the diagnosis of one or more conditions using a sample obtained from a human or animal body, the analyser comprising a first chamber having a first entry port and a second chamber having a second entry port, wherein each chamber is pre-loaded with assay reagents.

2. An analyser according to claim 1, wherein the first and second chambers are loaded with different assay reagents.

3. An analyser according to claim 1 or 2, wherein the first and second chambers each comprise an upper compartment and a lower compartment, separated by a breakable seal.

4. An analyser according to any of claims 1 to 3, wherein the upper compartments comprise liquid assay reagents and the lower compartments comprise solid or liquid assay reagents.

5. An analyser according to any of claims 1 to 4, wherein the assay reagents include one or more of a pH dependent or pH independent redox indicator, a chomophore or a fluorophore.

6. An analyser according to claim 3 or 4, wherein the assay reagents of the upper compartments are selected from one or more of buffer, dimethyl formamide, dimethyl sulphoxide and water.

7. An analyser according to any of claims 4 to 6, wherein the assay reagents of the lower compartment are selected from one or more of the following combinations: urea hydrogen peroxide/3,5 dihydroxy phenylalanine; Indigo disulphonic acid/potassium; carbonate; Indigo tetrasulphonic acid/potassium carbonate; hydrogen peroxide/o-phenylenediamine; Safranin T/potassium carbonate; Phenol Red in glycine- or ammonium formate-potassium chloride; or Alizarin Red S in potassium dihydrogen phosphate-potassium chloride.

8. An analyser according to any of claims 1 to 7, wherein the assay reagents react to indicate the presence or absence of a substance in the biological sample.

9. An analyser according to any of claims 1 to 8, further comprising a probe comprising: a first swab tip and a second swab tip; and an actuator configured to transition the probe between a sampling configuration and an analysis configuration, wherein in the sampling configuration the first swab tip and the second swab tip are located such that they form a single swab tip, and wherein in the analysis configuration the first swab tip and the second swab tip are located relatively further apart than in the sampling configuration such that they are operable as two separate swab tips.

10. A probe suitable for collecting biological samples from a human or animal body, the probe comprising a handle connected to a first swab tip and a second swab tip; wherein the probe further comprises an actuator configured to transition the probe between a sampling configuration and an analysis configuration,

wherein in the sampling configuration the first swab tip and the second swab tip are located such that they form a single swab tip,

and wherein in the analysis configuration the first swab tip and the second swab tip are located relatively further apart than in the sampling configuration such that they are operable as two separate swab tips.

11. A probe according to claim 10, wherein the probe is a single piece construction.

12. A probe suitable for collecting biological samples from a human or animal body, the probe comprising a handle and a first receiving port for a first swab having a first swab tip and a second receiving port for a second swab having a second swab tip; wherein the probe further comprises an actuator configured to transition the probe between a sampling configuration and an analysis configuration;

13. A probe according to any of claims 9 to 12, wherein the first swab tip and second swab tip are resiliently biased toward the sampling configuration.

14. A probe according to any of claims 9 to 13, wherein the actuator comprises a lever configured to pivot about a fulcrum to transition the probe between the sampling configuration and the analysis configuration.

15. A probe according to any of claims 9 to 13, wherein the actuator comprises a handle that is compressed to transition the probe between the sampling configuration and the analysis configuration.

16. A probe according to any of claims 9 to 15, wherein the first and second swab tips are substantially parallel when in the analysis configuration.

17. A probe according to claim 12, wherein the first and second receiving ports comprise grips for holding the first and second swabs.

18. A probe according to any of claims 9 to 17, wherein the probe comprises a chemical or biological agent for detecting a substance in the biological sample and a display for indicating the presence or absence of said substance in the sample.

19. A kit comprising:

an analyser according to any of claims 1 to 8 and a probe according to any of claims 10 to 18,

wherein the analyser is complementary in shape to the analysis configuration of the probe such that the first swab tip can be inserted via the first entry port into the first chamber simultaneously to the second swab being inserted via the second entry port into the second chamber.

20. A kit according to claim 19, further comprising instructions for use.

21. A kit according to claim 19 or 20 wherein the kit is in over the counter form.

22. A kit according to any of claims 19 to 20, further comprising one or more active therapeutic agents.

23. A kit according to claim 22, wherein the active agents are selected from at least two of an antibacterial agent, an antiviral agent and an antifungal agent.

24. A method of parallel diagnosis, the method comprising the steps of:

i) providing a biological sample simultaneously to the first and entry ports of the analyser of any of claims 1 to 8, and

ii) allowing the assay reagents to react with the biological sample in two parallel assays to provide two parallel output signals.

25. A method according to claim 24, wherein the biological sample is provided by a pipette or dropper.

26. A method according to claim 24, wherein the biological sample is provided by a probe according to any of claims 10 to 18.

27. A method according to claim 26, wherein the method comprises providing the biological sample simultaneously to the first and second swab tips of the probe, manipulating the actuator to transition the probe from the sampling configuration to the analysis configuration, and inserting the probe into the analyser such that the first swab tip is inserted via the first entry port into the first chamber simultaneously to the second swab tip being inserted via the second entry port into the second chamber.

28. A method of parallel diagnosis according to any of claims 24 to 27, wherein the method provides for simultaneous diagnosis of two different conditions.

29. A method of parallel diagnosis according to claim 28, wherein the conditions are selected from a bacterial infection, a viral infection and/or a fungal infection.

30. A method of parallel diagnosis according to claim 29, wherein the conditions are a bacterial infection and a fungal infection.

31. A method of parallel diagnosis according to claim 30, wherein the conditions are a Candida infection and a bacterial vaginosis infection.

32. A method of parallel diagnosis according to any of claims 24 to 31, wherein the parallel output signals are in the form of a colour change or an electronic reading

33. A method of parallel diagnosis according to any of claims 24 to 32, the method further comprising:

vii) administering a suitable treatment for the conditions identified.