Device for colleting menstrual blood

Abstract

The present invention relates to a self-sampling system for collecting menstrual blood comprising a collection device; and a container comprising a mixture of blood stabilizing substances for receiving the collected menstrual blood and to the use of said system for diagnosing a gynecological disease such as endometriosis.

Description

DEVICE FOR COLLETING MENSTRUAL BLOODDescriptionField of the Invention

[0001] The present invention relates to the field of a self-sampling system for collecting menstrual blood for the purpose of diagnosis of gynecological diseases. Background Art

[0002] Gynecological disorders refer to health issues in the female reproductive organs. These include the uterus, ovaries, fallopian tubes, external genitalia, and breasts. Some relatively common gynecological problems are bound to affect women’s health at some point in their lifetime. Most common gynecological disorders are inter alia uterine fibroids, vaginitis, polycystic ovaries syndrome (PCOS) and endometriosis.

[0003] Endometriosis is one of the most common gynecological diseases of the uterus, affecting up to 10% of all women of childbearing age worldwide. The cause is colonization of tissue that resembles the uterine lining outside the uterus. Symptoms of endometriosis include infertility and severe pain, especially during menstruation. As a result, endometriosis in general has a significant impact on quality of life and poses a threat to the physical, psychological and social integrity of many women. Diagnosis of endometriosis is difficult, since it often overlaps with other gynecological, urological and gastrointestinal symptoms, leading to long diagnostic delays. Diagnosis can take up to 7 years or even up to 8-12 years. The reasons for this may be patient-centered causes, such as shame, stigma, tolerance of or uncertainty about normal or abnormal symptoms, as well as physiciancentered causes, such as normalization of patient symptoms and delays in referral from primary care to a specialist. Finally, endometriosis is diagnosed after years by laparoscopy. Laparoscopy is a surgical procedure in which the abdomen is examined and endometriosis is histologically detected in the removed tissue.

[0004] A less invasive way to diagnose endometriosis would be the use of biomarkers. Thus, the use of diagnostic biomarkers would drastically shorten the time for diagnosing endometriosis. Additionally, it would be an improvement for women to have a point-of-care diagnostic device at hand for an early indication of a potential affliction with endometriosis.

[0005] Therefore, there is still the need for a point-of-care diagnostic device for early diagnosis of endometriosis.Summary of invention

[0006] It is the object of the present invention to provide a self-sampling system for collecting menstrual blood, wherein the system comprises a collection device and a container comprising a mixture of blood stabilizing substances and to the use thereof for diagnosing gynecological diseases. The object is solved by the subject matter of the present invention.

[0007] According to the invention, there is provided a self-sampling system for collecting menstrual blood comprising: a. a collection device; and b. a container comprising a mixture of blood stabilizing substances for receiving the collected menstrual blood.

[0008] Preferably, the mixture of blood stabilizing substances comprises at least one DNA stabilizing substance, and / or at least one RNA stabilizing substance, and / or at least one protease inhibitor, and / or at least one protein stabilizing substance, and / or a lysis buffer.

[0009] According to one embodiment of the invention, the collection device is comprised in a sanitary napkin, in a menstrual cup, or in a vaginal swab. The collection device may be a small container, a pad, an absorbent strip, or a cotton wad. When comprised in the sanitary napkin, the collection device may be covered by a semipermeable membrane penetrable by menstrual blood or configured as an inverse patch with a detachable absorbent strip. When configured as a vaginal swab, the collection device comprises a cotton wad for self-collection of menstrual blood. The vaginal swab may have a predetermined breaking point for detachment after insertion into the container containing the blood stabilizing substances.

[0010] When comprised in the menstrual cup, the collection device may be mounted to the menstrual cup via fasteners or screwed. When comprised in the sanitary napkin the collection device may be removable via a detachable film located on the bottom of the sanitary napkin.

[0011] A further embodiment relates to the collection device as described herein being configured to collect a defined volume of menstrual blood.

[0012] One embodiment of the invention relates to a method of diagnosing a gynecological disease, the method comprising the following steps: a. obtaining a sample of menstrual blood comprised in a collection device as described herein;b. transferring the menstrual blood sample into the container comprising the mixture of blood stabilizing substances; c. sealing the container comprising the stabilized menstrual blood sample; d. shipping the sealed container to a diagnostic center; and e. analyzing the stabilized menstrual blood sample in the laboratory to detect at least one biomarker for a gynecological disease.

[0013] According to an embodiment of the invention, the analysis may comprise performing quantitative measurement of at least one biomarker for a gynecological disease

[0014] A further embodiment relates to the method as described herein, wherein the gynecological disease is endometriosis.

[0015] One embodiment of the invention relates to a method for detecting biomarkers in a menstrual blood sample, the method comprising the following steps:1. obtaining a sample of menstrual blood comprised in a collection device as described herein;2. transferring the menstrual blood sample into the container comprising the mixture of blood stabilizing substances;3. sealing the container comprising the stabilized menstrual blood sample;4. shipping the sealed container to a laboratory; and5. measuring the presence and / or concentration of at least one biomarker in the stabilized menstrual blood sample.

[0016] According to an embodiment of the invention, the measurement may comprise performing quantitative measurement of at least one biomarker for a gynecological disease

[0017] A further embodiment relates to the method as described herein, wherein said at least one biomarker is associated with endometriosis.

[0018] One embodiment of the invention relates to a kit of parts for testing the presence of a biomarker for a gynecological disease in a menstrual blood sample comprising: a) a leaflet with directions for use; b) a system for collecting menstrual blood as described herein; and c) a shipping container for safe delivery of the sample to a diagnostic center.

[0019] A further embodiment relates to the kit of parts as described herein, wherein the leaflet provides instructions for: a) collecting the menstrual blood sample; b) transferring the sample to the container containing stabilizing substances; c) sealing the container; and d) shipping the container to the diagnostic center.

[0020] Preferably, wherein the collection device of the kit of parts is configured for self-use without any healthcare professional assistance

[0021] A further embodiment relates to the kit of parts as described herein, wherein the gynecological disease is endometriosisDescription of Embodiments

[0022] According to one embodiment of the invention there is provided a selfsampling system for collecting menstrual blood comprising a collection device and a container, wherein the collection device is configured to be comprised in a sanitary napkin, a vaginal swab, or a menstrual cup or to be connectable to a menstrual cup, and wherein said container comprises a mixture of blood stabilizing substances for receiving the collected menstrual blood. The container comprising the blood stabilizing substances may be separated from the collection device or the container and the collection device may be connected to each other.

[0023] The collection device may be configured as a small container, a pad, an absorbent strip, or a cotton wad.

[0024] The collection device may be comprised in a sanitary napkin. The collection device may be placed in the middle of the sanitary napkin with the opening covered by a semi-permeable membrane at the top. The collection device may be configured as a small, elongated vessel, wherein one end is open. The opening of the collection device is closed by the semi-permeable membrane. By wearing the sanitary napkin during menstruation for a specified wearing time, the collecting device fills with a defined volume of menstrual blood.

[0025] Alternatively, the collection device comprised in the sanitary napkin may be configured as an inverse patch with a detachable absorbent strip. The absorbent strip is configured to collect menstrual blood during use and is detachable for subsequent transfer to the container comprising the stabilizing substances.

[0026] The sanitary napkin exhibits a removeable film on the bottom side of the napkin. The collection device can be easily removed by stripping off the removeable film from the sanitary napkin.

[0027] Upon wearing the sanitary napkin during menstruation, a small, defined volume of the menstrual blood is collected in the collection device whereas the larger amount is collected in the sanitary napkin.

[0028] Alternatively, the collection device may be connected to a menstrual cup by suitable connection means.

[0029] A menstrual cup comprises a body part formed in a cup shape, a storage part which is comprised in the body part to store menstrual blood therein and a handle. The storage part is formed inside the body part of the menstrual cup to serve to store menstrual blood which is discharged during menstruation and acts as a collection device. The handle of the menstrual cup may be formed to protrude from a side opposite the opening of the body part and may serve to facilitate insertion of the menstrual cup into the vagina or removal of the menstrual cup therefrom.

[0030] Alternatively, the handle is configured to exhibit connection means in order to connect the collecting device to the menstrual cup. The collection device may be sealed by a semi-permeable membrane. The semipermeable membrane allows the menstrual blood to flow into the collecting device.

[0031] The connections means may be a screw, a click connection or any other suitable connection which allows an easy removal of the collection device after wearing.

[0032] Upon wearing the menstrual cup during menstruation, usually a large amount of menstrual blood is collected in the menstrual cup.

[0033] According to the invention, upon wearing the menstrual cup during menstruation, a small defined volume of the menstrual blood is collected in the collection device whereas the larger amount is collected in the storage part of the menstrual cup.

[0034] After the menstrual cup is taken out, the menstrual blood in the storage part of the menstrual cup is transferred to a separate container comprising a mixture of blood stabilizing substances.

[0035] Alternatively, after the menstrual cup is taken out, the collecting device is removed from the body part of the menstrual cup. The menstrual blood in thecollection device and / or in storage part can subsequently be transferred to the container comprising the stabilizing substances.

[0036] To this end, the menstrual cup may be formed of silicone having excellent elasticity and stretchability and no toxicity.

[0037] Alternatively, the collection device may be configured as a vaginal swab comprising a cotton wad for self-collection of menstrual blood. The vaginal swab may comprise a predetermined breaking point configured for detachment of the cotton wad after insertion into the container containing the blood stabilizing substances.

[0038] The transfer of menstrual blood from the vaginal swab to the container comprising stabilizing substances may be accomplished by either: a) rotating the swab in the stabilizing fluid for a predetermined duration and number of rotations and subsequently removing the swab, or b) breaking off the swab at the predetermined breaking point after insertion into the stabilizing fluid.

[0039] According to one embodiment of the invention, the container for receiving the collected menstrual blood comprises a defined volume of a blood stabilizing mixture. The blood stabilizing mixture may be present in an amount of about 0.05 to 5 mL. For example, the collecting device comprises about 0.05, 0.01, 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or about 5.0 mL of the blood stabilizing mixture.

[0040] The transfer process from the collection device to the stabilizing container is the following for each collection method: a) for the sanitary napkin comprising the collection devise having a semipermeable membrane, the collected sample is directly transferred to the container after removal of the collection device; b) for the inverse patch configuration, the detached absorbent strip is placed into the container containing stabilizing substances; c) for the menstrual cup collection device, the collected sample is transferred to the container manually from the storage part and / or after removal of the collection device from the menstrual cup; d) for the vaginal swab, either the rotation method or breaking method may be employed as described herein.

[0041] Each transfer method enables maintaining sample integrity and ensures appropriate collection.

[0042] The container comprising a mixture of blood stabilizing substances may be configured as a collection tube with a volume of approximately 1-10 ml. The container comprises a closure, for example a screw cap. Alternatively, the container comprises an attached lid. The container is specifically designed to receive and store the menstrual blood sample in the stabilizing solution until analysis.

[0043] When using the vaginal swab configuration of the collection device, the swab is inserted directly into the container comprising the stabilizing solution. The predetermined breaking point of the swab is positioned such that upon breaking, the cotton wad remains properly immersed in the stabilizing solution when the container is sealed.

[0044] Alternatively, when using the inverse patch configuration of the collection device, the detachable absorbent strip is removed from the sanitary napkin and inserted into the container comprising the stabilizing solution. The dimensions of the absorbent strip and the container are configured to ensure complete immersion of the strip in the stabilizing solution.

[0045] When using a conventional menstrual cup without an integrated collection device, a defined volume of menstrual blood is transferred from the menstrual cup into the container comprising the stabilizing solution using a disposable transfer device. The transfer device may include volume markings to ensure transfer of the correct sample volume. Alternatively, a small amount can be manually transferred from the storage part to the container comprising the stabilizing solution.

[0046] When using a collection device with a semipermeable membrane, whether integrated in a menstrual cup or in a sanitary napkin, the semipermeable membrane is removed after sample collection. The collected menstrual blood is then transferred into the container comprising the stabilizing solution. The membrane removal process is designed to maintain sample integrity during the transfer process.

[0047] The container may further comprise volume markings to indicate the proper ratio between the collected menstrual blood sample and the stabilizing solution. The container material is selected to be compatible with the stabilizing solution and to maintain sample integrity during storage and transport to the diagnostic center and / or laboratory.

[0048] The blood stabilizing mixture comprises at least one DNA stabilizing substance, and / or at least one RNA stabilizing substance, and / or at least one protease inhibitor, and / or a lysis buffer.

[0049] The stabilizing mixture may comprise a nuclease inhibitor that acts to prevent DNase and / or RNase activity within a blood sample. The nuclease inhibitor is preferably present in an amount sufficient to prevent a decrease in the amount and / or quality of the nucleic acids recoverable from the collected sample as compared with a sample that does not include a nuclease inhibitor. Nuclease inhibitors that may be used include, but are not limited to dithiothreitol (DTT), iodoacetamide, iodoacetic acid, heparin, chitosan, cobalt chloride, diethyl pyrocarbonate, ethanol, aurintricarboxylic acid (ATA), glyceraldehydes, sodium fluoride, ethylenediamine tetraacetic acid (EDTA), formamide, vanadylribonucleoside complexes, macaloid, hydroxylamine-oxygen-cupric ion, bentonite, ammonium sulfate, beta-mercaptoethanol, cysteine, dithioerythritol, tris(2- carboxyethyl) phosphene hydrochloride, a divalent cation such as Mg2+, Mn2+, Zn2+, Fe2+, Ca2+, Cu2+or any combination thereof.

[0050] The stabilizing mixture may also include one or more metabolic inhibitors in a suitable amount to reduce cell metabolism within a blood sample. Metabolic inhibitors that may be used include, but are not limited to glyceraldehyde, dihydroxyacetone phosphate, glyceraldehyde 3-phosphate, 1,3- bisphosphoglycerate, 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, pyruvate and glycerate dihydroxyacetate, sodium fluoride, K2C2O4, or any combination thereof.

[0051] The stabilizing mixture may also include one or more chelators capable of bonding with metal ions. The purpose of the one or more chelators is to further minimize nuclease activity and the resulting nucleic acid degradation. RNA cleavage via RNase activity requires the presence of divalent metal ions. The metal ion chelators will act by bonding with the metal ions thereby inactivating the ions and reducing the RNase effect of the metal ions. Possible metal ion chelators for addition to the protective agent include but are not limited to one or any combination of ethylene glycol tetraacetic acid (EGTA), l,2-bis-(o-Aminophenoxy)- ethane-N,N, — N' ,N' -tetraacetic acid tetraacetoxy-Methyl ester (BAPTA-AM), dietyldithiocarbamate (DEDTC), ethylenediaminetetraacetic acid (EDTA),dicarboxymethyl-glutamic acid, nitrilotriacetic acid (NTA), or ethylenediaminedisuccinic acid (EDDS).

[0052] The stabilizing mixture may also include a substance that acts to prevent oxidative stress. Nucleic acids and RNA in particular have been found to be highly susceptible to oxidative stress. Thus, the addition of antioxidants and / or reactive oxygen species (ROS) scavengers to the protective agent may help to protect the cellular RNA from the deleterious effects of oxidative damage. As used herein, the term “reactive oxygen species (ROS) scavenger group” refers to a group capable of acting as a scavenger of, or reacting with, superoxide (O2) or other reactive oxygen species (ROS) including hydroxyl radicals, peroxynitrite, hypochlorous acid and hydrogen peroxide. Additional examples of such antioxidants and reactive oxygen species scavengers include but are not limited to one or any combination of polyphenols such as flavonoids, phenolic acids, D-Mannitol, N-acetyl-L-cysteine, natural phenolic antioxidants (alpha-hydroxytyrosol, tyrosol, caffeic acid, alphatocopherol) as well as commercial phenolic antioxidants (BHT and BHA) or carotenoids.

[0053] The stabilizing mixture may also include one or more polycations (preferably polyamines) in a suitable amount such that they are capable of binding with any nucleic acids thereby preventing degradation of the nucleic acids. While polycations generally bind to nucleic acids, many polycations also alter cell membrane structure which may be associated with the loss of cell markers located on the cell membrane. Polyamines are naturally synthesized cations that do not compromise the structure of the cell membrane and thus are highly preferred for their ability to bind specifically to cellular RNA, based upon the polyanionic nature of the RNA. In binding to the RNA, the polyamines are able to protect the nucleic acids from RNase activity. The polyamines that may be added include but are not limited to protamine, spermine, spermidine, putrescine, cadaverine, or any combination thereof.

[0054] The stabilizing mixture may also include one or more protease inhibiting compounds for inhibiting enzyme activity that may have deleterious effects on the integrity of any nucleic acids present in a blood sample. These protease inhibiting compounds may include but are not limited to antipain, aprotinin, chymostatin, elastatinal, phenylmethylsulfonyl fluoride (PMSF), APMSF, TLCK, TPCK, leupeptin, soybean trypsin inhibitor, indoleacetic acid (IAA), E-64, pepstatin, VdLPFFVdL,EDTA, 1,10-phenanthroline, phosphoramodon, amastatin, bestatin, diprotin A, diprotin B, alpha-2-macroglobulin, lima bean trypsin inhibitor, pancreatic protease inhibitor, egg white ovostatin, egg white cystatin or any combination thereof. Combinations of protease inhibitors, commonly referred to as a “protease inhibition cocktail” by commercial suppliers of such inhibitors, may also be used as the stabilizing agent. Such “cocktails” may be generally advantageous in that they provide stabilization for a range of proteins of interest.

[0055] The stabilizing mixture may further include one or more phosphatase inhibitors for inhibiting enzyme activity that may have deleterious effects on the integrity of any nucleic acids present in a blood sample. These phosphatase inhibiting compounds may include but are not limited to calyculin A, nodularin, NIPP-1, microcystin LR, tautomycin, okadaic acid, cantharidin, calyculin A, microcystin LR, okadaic acid, fostriecin, tautomycin, cantharidin, endothall, nodularin, cyclosporin A, FK 506 / immunophilin complexes, cypermethrin, deltamethrin, fenvalerate, bpV(phen), dephostatin, mpV(pic) DMHV, sodium orthovanadate or any combination thereof.

[0056] The present stabilizing mixture may further include one or more protein stabilizing agents such as trehalose, mannitol, Tween 80, or polyvinyl alcohol.

[0057] The present stabilizing mixture may include one or more preservative agents, one or more metabolic inhibitors, one or more nuclease inhibitors and one or more metal ion chelators.

[0058] One embodiment of the invention relates to a method of diagnosing a gynecological disease, the method comprising the following steps: a) obtaining a sample of menstrual blood using a self-sampling system as described herein, b) transferring the menstrual blood sample from the collection device into the container comprising the mixture of blood stabilizing substances, c) sealing the container comprising the stabilized menstrual blood sample, d) shipping the sealed container to a diagnostic center, and e) analyzing the stabilized menstrual blood sample in the laboratory to detect at least one biomarker for a gynecological disease.

[0059] W02009068254A1 discloses endometrial secretion markers for the assessment of endometriosis. W02009068254A1 discloses a group of biomarkers that are considered to be of significant value for the detection, diagnosis, prognosisand / or monitoring of endometriosis: Said group of biomarkers comprises CTG1, SAHH, ADSS, NUDT5, AK1A1, AL1A3, AMY1, ANXA1 , AP0A1, ACTB, APOH, BPGM, CAT, CLIC4, CCDC94, AMPL, CAPZB, FRIH, FRIL, BLVRB, LEG1, GSTO1, GSTP1, PAEP, LGUL, HSPB1, HBB, LEI, MSN, PPIA, PGAM1, PMVK, PCBP1, PNPO, PSB3, PSD7, PSD10, 1433B, 1433G1;1433S, 1433T, 1433Z, ARHGDIA, GD15, PRPS2, SEP11, NEK7, SAS, TCTP, TBB5 and WDR1 or their transcriptional or post-translational variants, and combinations thereof.

[0060] According to a further embodiment of the invention, there is provided a kit of parts for testing the presence of a biomarker for a gynecological disease comprising: a) a leaflet with directions for use, b) a self-sampling system for collecting menstrual blood as described herein, and c) a shipping container for safe delivery of the sample to a diagnostic center.

[0061] The leaflet provides instructions for: a) collecting the menstrual blood sample using the collection device of the self-sampling system, b) transferring the sample to the container containing stabilizing substances, c) sealing the container, and d) shipping the container to the diagnostic center.

[0062] The kit of parts is specifically configured for self-use at home. Thus, no healthcare professional is required for collecting and preparing the sample for analysis.The gynecological disease may be endometriosis, adenomyosis, polycystic ovary syndrome (PCOS), endometrial hyperplasia, uterine and / or endometrial cancer, cervical cancer, ovarian cancer, and / or cervical dysplasia.

Claims

Claims1. A self-sampling system for collecting menstrual blood comprising: a. a collection device; and b. a container comprising a mixture of blood stabilizing substances for receiving the collected menstrual blood.

2. The self-sampling system according to claim 1, wherein the mixture of blood stabilizing substances comprises at least one DNA stabilizing substance, and / or at least one RNA stabilizing substance, and / or at least one protease inhibitor, and / or at least one protein stabilizing substance, and / or a lysis buffer.

3. The self-sampling system according to claim 1 or 2, wherein the collection device is comprised in a sanitary napkin, in a menstrual cup, or in a vaginal swab.

4. The self-sampling system according to any one of claims 1 to 3, wherein said collection device is a small container, a pad, an absorbent strip, or a cotton wad.

5. The self-sampling system according to any one of claims 1 to 4, wherein the collection device comprised in the sanitary napkin is covered by a semipermeable membrane penetrable by menstrual blood.

6. The self-sampling system according to any one of claims 1 to 4, wherein the collection device comprised in the sanitary napkin is an inverse patch with a detachable absorbent strip.

7. The self-sampling system according to any one of claims 1 to 4, wherein the collection device is vaginal swab comprising a cotton wad for self-collection of menstrual blood.

8. The self-sampling system according to any one of claims 1 to 4, wherein the collection device comprised in a menstrual cup is mounted to said menstrual cup via fasteners or screwed.

9. The self-sampling system according to any one of claims 1 to 4, wherein the collection device comprised in the sanitary napkin is removable via a detachable film located on the bottom of the sanitary napkin.

10. The self-sampling system according to any one of claims 1 to 4, wherein the vaginal swab comprises a predetermined breaking point configured for detachment of the cotton wad after insertion into the container containing the blood stabilizing substances.

11. The self-sampling system according to any one of claims 1 to 10, wherein the collection device is configured to collect a defined volume of menstrual blood.

12. A method of diagnosing a gynecological disease, the method comprising the following steps: a. obtaining a sample of menstrual blood comprised in a collection device according to any one of claims 1 to 11; b. transferring the menstrual blood sample into the container comprising the mixture of blood stabilizing substances; c. sealing the container comprising the stabilized menstrual blood sample; d. shipping the sealed container to a diagnostic center; and e. analyzing the stabilized menstrual blood sample in the laboratory to detect at least one biomarker for a gynecological disease.

13. The method according to claim 12, wherein analyzing the stabilized menstrual blood sample comprises performing quantitative measurement of at least one biomarker for a gynecological disease.

14. The method according to claim 12 or 13, wherein the gynecological disease is endometriosis.

15. A method for detecting biomarkers in a menstrual blood sample, the method comprising the following steps: a. obtaining a sample of menstrual blood comprised in a collection device according to any one of claims 1 to 11; b. transferring the menstrual blood sample into the container comprising the mixture of blood stabilizing substances; c. sealing the container comprising the stabilized menstrual blood sample; d. shipping the sealed container to a laboratory; and e. measuring the presence and / or concentration of at least one biomarker in the stabilized menstrual blood sample.

16. The method according to claim 15, wherein measuring comprises performing quantitative measurement of said at least one biomarker.

17. The method according to claim 15 or 16, wherein said at least one biomarker is associated with endometriosis18. A kit of parts for testing the presence of a biomarker for a gynecological disease in a menstrual blood sample comprising: d) a leaflet with directions for use;e) a system for collecting menstrual blood according to any one of claims 1 to 11; and f) a shipping container for safe delivery of the sample to a diagnostic center.

19. The kit of parts according to claim 18, wherein said leaflet provides instructions for: e) collecting the menstrual blood sample; f) transferring the sample to the container containing stabilizing substances; g) sealing the container; and h) shipping the container to the diagnostic center.

20. The kit of parts according to claim 18 or 19, wherein the collection device is configured for self-use without any healthcare professional assistance.

21. The kit of parts according to any one of claims 19 to 20, wherein the gynecological disease is endometriosis.