Methods for detecting prostate cancer biomarkers

Abstract

The present disclosure relates to a method for detecting a prostate cancer related biomarker, comprising contacting a diluted sample with a modulator selected from the group consisting of sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazene]naphthalene-1-sulfonate, sodium 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazene]-7-nitronaphthalene-1-sulfonate, triisopropylsilane, and iron(III) chloride, and a luminescent label to obtain a measurement sample. Then, the measurement sample is incubated for a period of time and subsequently excited. The time-resolved luminescence signal of the label in the measurement sample is measured, and if the luminescence signal is at least 50% higher than the luminescence signal of a control sample, the control sample being from a human subject not suffering from prostate cancer, the likelihood of prostate cancer in the human subject is increased.

Description

TECHNICAL FIELD

[0001] The present disclosure relates to a method for detecting prostate cancer related biomarkers from a urine sample. The present disclosure also relates to a pre-screening method and kit that can be used at home and to a system for the automated analysis of a urine sample. BACKGROUND

[0002] Prostate cancer is the most common type of cancer in men. Over-diagnosis of prostate cancer can lead to over-treatment. This often results in long-term side effects that affect the quality of life. The challenge is to distinguish between very slow growing and least harmful types of prostate cancer and aggressive metastatic types of prostate cancer. Typical prostate cancer diagnosis includes prostate specific antigen (PSA) testing, digital rectal examination, cystoscopy and magnetic resonance imaging (MRI) methods. PSA testing is the simplest and most cost-effective as it can be measured in a laboratory from a blood sample. However, the disadvantage of PSA testing is that the amount of PSA in the blood can also be elevated for several other reasons (e.g. under benign conditions) and thus the test can produce a higher number of false positive results. Such false positive results lead to additional diagnosis and unnecessary treatment, not to mention mental stress. None of the above listed methods are suitable for population level prostate cancer screening. SUMMARY

[0003] In view of these known diagnostic methods, there is a need for an economic, simple, fast and reliable method to diagnose aggressive prostate cancer. Advantageously, it is also aimed to provide a method for pre-screening prostate cancer at home using a simple test kit. Furthermore, it is still aimed to provide an automated screening system that is easy to implement.

[0004] Typical methods for detecting prostate cancer related biomarkers include:

[0005] - contacting a urine sample of a human subject with a conditioning agent selected from the group consisting of 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonic acid sodium salt, 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonic acid sodium salt, triisopropylsilane, and iron (III) chloride; and a luminescent tag to obtain a measurement sample;

[0006] - incubating the measurement sample;

[0007] - illuminating the measurement sample with excitation light; and

[0008] - measuring the time-resolved luminescence signal of the label in the measurement sample and determining an increased likelihood of prostate cancer in the human subject if the luminescence signal is at least 50% higher than the luminescence signal of a control sample, the control sample being from a human subject not suffering from prostate cancer.

[0009] Another typical method for pre-screening prostate cancer related biomarkers comprises:

[0010] - contacting a urine sample of a human subject in a test tube, wherein the test tube comprises triisopropylsilane and 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonic acid sodium salt;

[0011] - incubating the test tube with the sample; and

[0012] - visually observing the test tube and determining an increased likelihood of prostate cancer if a color change of the content of the test tube is observed.

[0013] An atypical kit of parts comprises

[0014] - a test tube comprising triisopropylsilane and 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonic acid sodium salt;

[0015] - means for collecting a urine sample; and

[0016] - means for contacting the diluted urine sample with the test tube.

[0017] A typical system for automatically analyzing a urine sample, the system being connectable to a toilet and comprising:

[0018] - at least one detection device configured to receive a urine sample, to detect an initial color of the urine sample, to bring the urine sample in contact with a conditioning agent to form a measurement sample, to incubate the measurement sample and to detect a color after incubation and / or an intensity of the color;

[0019] - a computing device configured to analyze the detected color and optionally the color intensity and to send information of any detected increased risk of prostate cancer; and

[0020] - means for transferring the detected color and optionally the color intensity to the computing device. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Results of experiments are shown.

[0022] Figures 2A-2FFurther experimental results are shown.

[0023] Figure 3A and 3B is a schematic representation of an exemplary system for detecting prostate cancer. DETAILED DESCRIPTION

[0024] The method for detecting prostate cancer related biomarkers according to one embodiment comprises:

[0025] - contacting a urine sample of a human subject with a modulator selected from the group consisting of sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazene]naphthalene-1- sulfonate, sodium 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazene]-7-nitronaphthalene-1- sulfonate, triisopropylsilane, and iron (III) chloride, and a luminescent tag to obtain a measurement sample;

[0026] - incubating the measurement sample;

[0027] - irradiating the measurement sample with excitation light; and

[0028] - measuring the time-resolved luminescence signal of the tag in the measurement sample, and if the luminescence signal is at least 50% higher than the luminescence signal of a control sample, the control sample being from a human subject not suffering from prostate cancer, determining an increased likelihood of prostate cancer of the human subject.

[0029] The present disclosure thus provides an economic but reliable and fast method for the pre-screening and early detection of malignant changes related to prostate cancer, as it enables the detection of prostate cancer related biomarkers as demonstrated in the experimental section below. Currently, the accuracy of this test method is believed to be at least 70-80%, while the accuracy of the above-mentioned conventional PSA test is significantly lower. Thus, the number of false positives is significantly reduced compared to the traditional method. The method is thus accurate and the test is reliable.

[0030] The sample preparation of the method takes less than 5 minutes, while the test itself takes about 10 minutes. The method is thus used efficiently and does not require any blood sample, which further reduces the cost of the test as less health care personnel is needed.

[0031] It is believed that the present method can also provide more information about early benign changes that can lead to the development of cancer. In fact, as it is a pre-screening method with high accuracy, it is easy to use and can also be used at home (see below), thus enabling easier monitoring of patients to detect changes that can cause trouble later on. In addition to pre-screening, the present method can also be used in cancer research including cellular and genetic research systems.

[0032] It is believed that the method is based on the link between cancer and iron metabolism. This link is known, but its role in cancer physiology is unknown (Wang Y, Yu L, Ding J, Chen Y. Iron Metabolism in Cancer. Int JMol Sci. 2018; 20(1): 95.). Urinary iron concentration is usually between 50-100 pg / L, varying with individual physiology and age (Pfrimer, Karina et al. "Impact of aging on urinary excretion of iron and zinc" Nutrition and metabolic insights (2014) vol. 7; 47-50. 26.). The method is based on the difference in iron-binding molecules in urine samples from healthy and unhealthy subjects. In fact, these molecules are more abundant in cancer samples than in healthy samples. This difference is detected by the described method. It is assumed that the iron concentration is the same in cancerous and healthy samples, but the amount of iron-binding agent is different. Therefore, the signal is higher in cancer cases, because free iron is taken up by the binding agent. In healthy samples, iron remains in the sample and interferes with the label, so that the luminescence signal is reduced.

[0033] The modulator 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazene]- naphthalene-1-sulfonic acid sodium salt is sold under the trade name Calcon TM (CAS No. 2538-85-4) and the modulator 3-hydroxy-4-[(1-hydroxynaphthalen-2- yl)diazene]-7-nitronaphthalene-1-sulfonic acid sodium salt is sold under the trade name Calmagite® Black T (CAS No. 1787-61-7). Iron (III) chloride (CAS No. 7705-08-) with a purity of 97% and triisopropylsilane (CAS No. 6485-79-6) with a purity of 98% are preferred.

[0034] When 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazene]-naphthalene-1- sulfonic acid sodium salt is used as modulator, it is usually used in an amount of 50-800 pM, most usually 250 pM, in a volume of 4 mI. When 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazene]-7-nitronaphthalene-1-sulfonic acid sodium salt is used as modulator, it is usually used in an amount of 50-1000 pM, preferably 500 pM, in a volume of 4 mI. When triisopropylsilane is used as modulator, it is usually used in an amount of 4 mI of a 10% solution, but solutions of 2-25% can also be used. When iron (III) chloride is used as modulator, it is usually used in an amount of 5-500 pM, preferably 300 pM.

[0035] In the present method, the likelihood of prostate cancer in a human subject is determined to be increased if the luminescence signal is at least 50% higher than the luminescence signal of a control sample, said control sample being from a human subject not suffering from prostate cancer. Thus, the luminescence signal can be at least 50, 75, 90, 100, 125, 150, 175, 200, 250 or 300% higher than the control sample, or even higher.

[0036] Generally, the present method does not comprise steps performed on a human body.

[0037] The label used can be an Europium chloride (EuCl 3+ ) label or a Terbium chloride (TbCl 3+ ) label. As known to the person skilled in the art, these labels are present in the form of a complex. For example, Europium chloride (III) 99.99%, CAS number 10025-76-0 from Sigma-Aldrich, and Terbium chloride (III) 99.99%, CAS number 10042-88-3 from Sigma-Aldrich can be used. The Europium chloride label can be used with a complex of, for example, Europium chloride, nitrilotriacetic acid (NTA) (e.g. 99%, CAS number 139-13-9 from Sigma-Aldrich) and trioctylphosphine oxide (TOPO) (e.g. 99%, CAS number 78-50-2 from Sigma-Aldrich). One possible combination is to mix these three ingredients in a ratio of 3:9:9. Instead of NTA, 2-Thiophenecarbonyltrifluoroacetone (e.g. 99%, CAS number 326-91-0 from Sigma-Aldrich) or 4,4,4-Trifluoro-1-(2-furyl)-1,3-butanedione (e.g. 99%, CAS number 326-90-9 from Sigma-Aldrich) can also be used.

[0038] The concentrations of the various components of the europium chloride complex can be 80 nM - 5.0 μM for the europium chloride; 5 nM - 3 μM for the TOPO; and 100 nM - 3 μM for the NTA. Thus the concentration of the europium chloride can be, for example, 80 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, or 4 μM up to 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, 3 μM, 3.5 μM, 4 μM, 4.5 μM, or 5 μM. The concentration of the TOPO can be, for example, 5 nM, 10 nM, 15 nM, 20 nM, 25 nM, 30 nM, 35 nM, 40 nM, 45 nM, 50 nM, 55 nM, 60 nM, 65 nM, 70 nM, 75 nM, 80 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, or 2 μM up to 15 nM, 20 nM, 25 nM, 30 nM, 35 nM, 40 nM, 45 nM, 50 nM, 55 nM, 60 nM, 65 nM, 70 nM, 75 nM, 80 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, or 3 μM. Thus the concentration of the NTA can be, for example, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, or 2 μM up to 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 1.5 μM, 2 μM, 2.5 μM, or 3 μM.

[0039] According to one embodiment, the tag is used in an amount of 4 μl per microtiter well. Thus, one possibility is to use the tag in an amount of 4 μL of a tag mixture containing europium chloride 0.717 μM in dimethyl sulfoxide, trioctylphosphine oxide 0.430 μM, and nitrilotriacetic acid 0.430 μM (NTA) in dimethyl sulfoxide.

[0040] The excitation light can come from, for example, a pulsed laser, a light emitting diode (LED), or a xenon flash lamp.

[0041] According to one embodiment, the time-resolved luminescence signal is measured for a time of 200-800 ps after the delay time of 200-800 ps. Thus, the signal can be measured, for example, from 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 ps up to 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, or 800 ps. Independently thereof, the delay time can be, for example, from 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 ps up to 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, or 800 ps.

[0042] The urine sample can optionally be diluted before contacting the sample with the adjusting agent. The diluent and the adjusting agent can also be added simultaneously. The diluent can be, for example, a physiological salt solution. The dilution rate can be, for example, 1 :5-1 :20. A dilution rate of 1 :5 means that there is 1 volume of the sample and 5 volumes of the physiological salt solution. The dilution rate can be, for example, from 1 :5, 1 :6, 1 :7, 1 :8, 1 :9, 1 :10, 1 :11, 1 :12, 1 :13, 1 :14, 1 :15, or 1 :16 up to 1 :7, 1 :8, 1 :9, 1 :10, 1 :11, 1 :12, 1 :13, 1 :14, 1 :15, 1 :16, 1 :17, 1 :18, 1 :19, or 1 :20.

[0043] A typical incubation time is 1-15 minutes. Longer incubation times can reduce the efficiency of the method. The incubation time can be, for example, from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, or 12 minutes up to 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, or 15 minutes. The incubation is typically carried out at room temperature and under normal atmosphere.

[0044] The present disclosure also relates to another method, namely a method for pre-screening a prostate cancer related biomarker, comprising:

[0045] - contacting a urine sample of a human subject in a test tube, wherein the test tube comprises triisopropylsilane and 3-hydroxy-4-[(2-hydroxynaphthalen-1 -yl)diazenyl]naphthalene-1 -sulfonic acid sodium salt;

[0046] - incubating the test tube with the sample; and

[0047] - visually observing the test tube and determining an increased likelihood of prostate cancer if a color change of the content of the test tube is observed.

[0048] The method is thus associated with a test that can be performed at home by the subject. Unless otherwise indicated, the above disclosed embodiments and variants associated with the method using the label apply mutatis mutandis to this home method.

[0049] Most often, if a sample dilution method is used, the urine sample is diluted 1 :2 (1 part sample, 2 parts physiological saline solution). Both conditioners are placed in the sample tube. If the sample contains the cancerous marker, the color in the test tube will change to a color visible in the range of 630-650 nm, i.e. the test tube will turn red / dark red. For a healthy subject, there is no change in color. It is noted that the color remains in the test tube even if the sample is subsequently dried.

[0050] The change in color can be observed visually by the human eye or can be read by means of a specific application program, for example, installed on a mobile phone and used with the camera of the mobile phone.

[0051] The volume of the test tube is typically 1-10 ml, for example 3 ml. The dilution of the sample is preferably performed in the test tube. In this method, the incubation time is also typically 1-15 minutes, and the above listed variants apply mutatis mutandis. According to one embodiment, the amount of triisopropylsilane is 200 μΐ of a 10% solution for a 3 ml sample volume. Triisopropylsilane can also be used as a 5-25% solution. The amount of sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1 -sulfonate is 200 μΐ of a 250 μΜ solution for a 3 ml volume of the diluted sample. This conditioner can also be used in the range of 100 μΜ - 800 μΜ.

[0052] The present disclosure still further relates to a kit of parts comprising:

[0053] - a test tube comprising triisopropylsilane and sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1 -sulfonate;

[0054] - a device for collecting a urine sample; and

[0055] - a device for contacting the diluted urine sample with the test tube.

[0056] The kit of parts can thus be used for testing, for example, at home, in particular where there is less or no laboratory equipment available. The kit of parts also comprises, for example, in the form of an ampoule, some physiological saline solution for dilution. The kit of parts can still further comprise a device for diluting the urine sample.

[0057] The device for collecting the urine sample can be a sterile cup and a sterile syringe. The device for diluting the urine sample can comprise a sterile cuvette or a sterile tube made of a transparent material and to be marked with a marking showing the user how much physiological saline solution to add. The device for contacting the diluted urine sample with the test tube can be, for example, a pipette. The kit preferably further comprises instructions for use.

[0058] The present disclosure still further relates to a kit comprising the modulator and instructions for use thereof with a time-resolved luminescence measuring instrument, i.e. instructions for carrying out the method for determining the likelihood of prostate cancer. The kit can further comprise a database of control samples, a license to use the database, or some control samples for comparison. Furthermore, the kit can comprise a label.

[0059] The present disclosure also relates to the use of 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonic acid sodium salt, 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonic acid sodium salt, triisopropylsilane, or iron (III) chloride for the prescreening of prostate cancer. The use is typically ex vivo.

[0060] The present disclosure still further relates to a system for automatically analyzing a urine sample, which can be connected to a toilet. The system comprises:

[0061] at least one detection device configured to receive a urine sample, to detect the initial color of the urine sample, to bring the urine sample in contact with a modulator to form a measurement sample, to incubate the measurement sample and to detect the color after incubation and / or the intensity of the color;

[0062] a computing device configured to analyze the detected color and optionally the color intensity, and to send any detected information of an increased risk of prostate cancer; and

[0063] means for transferring the detected color and optionally the color intensity to the computing device.

[0064] The detection device can be attached to the interior of the toilet, for example to the front and / or the back of the toilet, so that a urine sample can be provided while urinating. For example, when two detection devices are used, they can be configured to wake up as soon as a urine sample is received and thereby only communicate when active (or awake). The detection device is preferably attachable by a removable attachment that does not require drilling or the like. The detection device can also be made integral with the toilet.

[0065] The means for transmitting the detected color and / or color intensity to the computing device can for example be by radio communication or similar. The computing device can also be connected to a server system, for example via a communication network, in which case the server system can perform the analysis and further actions based on any detected information indicating an increased risk of prostate cancer. The server can also be configured to store any information provided.

[0066] The analysis of the detected color and optionally color intensity comprises comparing the detected color before addition of the adjusting agent with the color or the intensity of the color after incubation. The color can also be detected immediately after addition of the adjusting agent, in which case both detections are based on the intensity of the color. The result of this comparison is then compared with results stored in a database, i.e. a database of results from samples of people not having prostate cancer and results from samples of people having prostate cancer. At least when indicating an increased risk of prostate cancer, the result of the latter comparison is then further communicated.

[0067] The detection device can also be configured to dilute the sample, for example using water from the toilet water system.

[0068] The detection device can have for example the following structure. It comprises an opening for receiving the urine sample, and the opening is connected to a first part of a fluid system. In this first part, water is also added to the sample, typically from the toilet water system, to form a diluted sample. The first part is at least partly transparent so that a sensor can perceive the diluted sample. The sensor can for example be a camera or a photodetector such as a photodiode, arranged to measure the color and / or intensity of light at 620-650 nm. Thus, the sensor reads the color of the diluted sample after it has been formed, typically within a few seconds, and transmits the measurement to a computing device. Thereafter, the diluted sample is contacted with a mixture of adjusting agents, as disclosed above in relation to the test method that can be performed at home, and incubated for 1-10 minutes, for example 5 minutes. During this time, the opening is closed so that more fluid cannot enter the detection device. This can be in the same part where dilution takes place, or in a different part. After incubation, the sample can again be kept in the same part where dilution took place, or moved to another part of the fluid system. If the sample is moved, the system can comprise a second sensor, or if the sample is kept in the same space, the first sensor is used again to measure the color and / or intensity of light emitted at 620-650 nm. Again, the result is transmitted to the computing device, which then performs the analysis (or for example sends the information further to a server), and if needed, for example sends an alarm to the mobile device of the user of the toilet.

[0069] Experimental part

[0070] Urine samples from known healthy subjects and known subjects suffering from prostate cancer were tested as follows.

[0071] Preliminary experiments were performed using 10 cancer and 12 healthy urine samples from human subjects. Before further steps, the samples were centrifuged at 10000 rpm for 5 min to remove any excess solid material. 1 ml of the clear supernatant was diluted to 9 ml physiological salt solution.

[0072] First, 4 μl of a modulator solution was added to the microtiter well. The modulator consisted of one of four different aforementioned modulators in MQ water. All modulators were obtained from Sigma-Aldrich. Then, each sample was divided into 3 parallel samples of 100 μl volume and pipetted into a 96 well plate.

[0073] Finally, 4 μl of a label mix was added to each microtiter well, the label mix containing Europium chloride 0.717 μM, Trioctylphosphine oxide (TOPO) 0.430 μM and Nitrilotriacetic acid 0.430 μM (NTA) in dimethylsulfoxide (DMSO).

[0074] After 10 minutes of incubation, the luminescence emission intensity was measured using a Victor 2 multilabel counter (Wallac, Perkin-Elmer Life and Analytical Sciences) in a 400 μs window after a 400 μs delay time.

[0075] Figure 1 Time resolved luminescence signals from samples of healthy subjects and subjects suffering from cancer are shown. Time resolved luminescence is given on the ordinate and time on the abscissa. The solid line represents the results from samples of healthy subjects ("normal" samples) and the dashed line represents the results from samples of subjects suffering from prostate cancer. Thus, there is a clear difference in luminescence, especially after a certain time.

[0076] Figures 2A-2F Luminescence signals from samples of healthy subjects and cancer subjects at different time points are shown, the height of the columns representing the luminescence. The number of tested samples of healthy subjects was 12 (1A, 1B, 1C, 1D, 1E, 1F, 1G, 1H, 1I, 1J, 1K and 1L) and the number of tested samples of cancer subjects was 10 (2A, 2B, 2C, 2D, 2E, 2F, 2G, 2H, 21 and 2J), each vertical column representing one sample. At time 0, the reagents listed above were added to the samples and at 0 minutes (1A, 1B, 1C, 1D, 1E, 1F, 1G, 1H, 1I, 1J, 1K, 1L, 2A, 2B, 2C, 2D, 2E, 2F, 2G, 2H, 2I and 2J), 5 minutes (1H, 1I, 1J, 1K, 1L, 2H, 2I and 2J) and 10 minutes (1I, 1J, 1K, 1L, 2I and 2J), the luminescence was measured. Figure 2A , 2C Figure 2B , 2D At time 0, the reagents listed above were added to the samples and at 0 minutes (1A, 1B, 1C, 1D, 1E, 1F, 1G, 1H, 1I, 1J, 1K, 1L, 2A, 2B, 2C, 2D, 2E, 2F, 2G, 2H, 2I and 2J), 5 minutes (1H, 1I, 1J, 1K, 1L, 2H, 2I and 2J) and 10 minutes (1I, 1J, 1K, 1L, 2I and 2J), the luminescence was measured. Figure 2A , 2B Figure 2C , 2D , Figure 2E ​​and 2F The luminescence is measured at the time of (i.e. immediately or after 5 or 10 minutes of incubation) addition of the substrate (i.e. luciferin, luciferase and ATP). The measurements at the 5 minute time point indicate that the signal levels of healthy samples decrease faster than cancer samples. After 10 minutes of incubation the signal difference is already significant and sufficient for diagnostic purposes.

[0077] Figure 3A is a schematic view of a toilet 300 seen from above. A detection device 310A is attached at the back side of the toilet 300, inside the toilet, and a second detection device 310B is attached at the front side of the toilet 300, inside the toilet. The detection devices 310A, 310B are attached in a way that they can provide a urine sample at the time of urination. The detection devices 310A, 310B are connected by radio communication to a computing device 340. The computing device 340 is in turn connected to a server system 344 by a communication network 342. The detection devices are configured to measure a visual signal from the urine sample and send the measurement to the server system 344 for further analysis and storage.

[0078] Figure 3B is an illustration of details of the detection device 310A in use. The detection device 310A is attached to the side 302 of the toilet by attachment means 314. The detection device 310A comprises a first opening 330 for receiving a urine sample and receiving water 306 from the water outlet 304 to dilute the sample. The urine sample and water are guided from the first opening 330 to a first portion 332 of the fluidic channel. The first portion 332 is at least partially transparent to a first sensor 320 to measure a first color of the diluted sample and provide the measured first color of the sample to a controller 324. The diluted sample is further guided through a second portion 334 of the fluidic system to a third portion 336 of the fluidic system. A conditioning agent is also supplied to the third portion 336 (not shown). The third portion 336 is configured to hold the diluted sample together with the conditioning agent for incubation. The third portion is at least partially transparent to a second sensor 322 to measure a second color and / or intensity thereof and provide the measured second color and / or intensity to the controller 324. As shown in Figure 3A the controller is connected to a communication module 326 and the communication module 326 provides the measurements to the computing device 340.

Claims

1. Use of a modulator and a luminescent label in preparing a kit for detecting prostate cancer-related biomarkers, comprising: - contacting a urine sample of a human subject with a modulator and a luminescent label to obtain a measurement sample, wherein the modulator is selected from the group consisting of sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonate, sodium 3-hydroxy-4-[(1-hydroxynaphthalen-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonate, triisopropylsilane, and iron(III) chloride; - incubating the measurement sample; - irradiating the measurement sample with excitation light; and - measuring a time-resolved luminescence signal of the label in the measurement sample, and determining an increased likelihood of prostate cancer in the human subject if the luminescence signal is at least 50% higher than a luminescence signal of a control sample, the control sample being from a human subject not suffering from prostate cancer; Wherein, the luminescent label is selected from europium chloride and terbium chloride.

2. The use according to claim 1, wherein After a delay time of 200-800 μs, the time-resolved luminescence signal was measured for a period of 200-800 μs.

3. The use according to claim 1 or 2, comprising diluting the urine sample in a physiological salt solution to obtain a diluted sample before contacting the sample with the conditioning agent, wherein The urine samples were diluted using a dilution ratio of 1:5-1:

20.

4. The use according to claim 1 or 2, wherein The regulator is sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonate, and the dosage is 50-800 μM.

5. The use according to claim 1 or 2, wherein The regulator is sodium 3-hydroxy-4-[(1-hydroxynaphthalene-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonate, and the dosage is 50-1000 μM.

6. The use according to claim 1 or 2, wherein The conditioning agent is triisopropylsilane, and the amount used is 4 μL of a 2-25% solution.

7. The use according to claim 1 or 2, wherein The regulator is iron (III) chloride, and the amount used is 5-500 μM.

8. The use according to claim 1 or 2, wherein The incubation time is 1-15 minutes.

9. Use of triisopropylsilane and sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonate in the preparation of a kit for pre-screening prostate cancer-related biomarkers, comprising: - contacting a urine sample from a human subject in a test tube, wherein the test tube comprises triisopropylsilane and sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonate; - incubating the test tube with the sample; and - visually observing the test tube, and if a color change of the contents of the test tube is observed, determining an increased likelihood of prostate cancer.

10. The use according to claim 9, wherein The incubation time is 1-15 minutes.

11. A kit of parts comprising: - a test tube containing triisopropylsilane and sodium 3-hydroxy-4-[(2-hydroxynaphthalen-1-yl)diazenyl]naphthalene-1-sulfonate; - a device for collecting a urine sample; and - means for contacting a diluted urine sample with said test tube.

12. Use of a modulator and a luminescent label in the preparation of a kit for pre-screening of prostate cancer, wherein the modulator is selected from sodium 3-hydroxy-4-[(2-hydroxynaphthalene-1-yl)diazenyl]naphthalene-1-sulfonate, sodium 3-hydroxy-4-[(1-hydroxynaphthalene-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonate, triisopropylsilane or iron (III) chloride; and the luminescent label is selected from europium chloride and terbium chloride.

13. The use according to claim 12, wherein The use is ex vivo.

14. A system for automatically analyzing a urine sample, the system being connectable to a toilet and comprising: - at least one detection device configured to receive a urine sample, detect the initial color of the urine sample, contact the urine sample with a modifier and a luminescent label to form a measurement sample, incubate the measurement sample and detect the color and / or the intensity of the color after incubation, wherein the modifier is selected from the group consisting of 3-hydroxybenzoic acid and benzophenone -4-[(2-hydroxynaphthalene-1-yl)diazenyl]naphthalene-1-sulfonate sodium, 3-hydroxy-4-[(1-hydroxynaphthalene-2-yl)diazenyl]-7-nitronaphthalene-1-sulfonate sodium, triisopropylsilane and iron (III) chloride, the luminescent label is selected from the group consisting of europium chloride and terbium chloride; - a computing device configured to analyze the detected colors and optionally the color intensities and to transmit information on any detected increased risk of prostate cancer; and - means for communicating said detected color and optionally color intensity to said computing device.

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