Use of a condom after sexual intercourse in the simultaneous collection and / or detection of male and female genital tract pathogens

Abstract

The present invention relates to the application of a condom after sexual life in simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts, belonging to the technical field of disease detection. The present invention uses a condom to collect reproductive tract pathogens during sexual life. The collection method is gentler, non-invasive, more comprehensive, with a high acceptance rate by users. It can simultaneously collect pathogens in the male and female reproductive tracts, and can collect pathogen samples that cannot be collected by ordinary reproductive tract swabs or need to be collected multiple times, which can increase the types and concentrations of collected pathogens and improve the detection accuracy of pathogens. At the same time, the present invention discovers that using a condom containing microsphere lubricant can more effectively enrich reproductive tract cells and increase the concentration of collected pathogens; using the cell preservation solution with specific components of the present invention can improve the preservation efficiency of the cells collected by the condom, improve the stability of the collected samples, and further improve the accuracy of pathogen detection.

Description

Technical Field

[0001] The present invention relates to the technical field of disease detection, and particularly to the application of a condom after sexual intercourse in simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts. Background Art

[0002] Reproductive tract infections can cause various diseases. Relatively common reproductive infections include gonorrhea, syphilis, non-gonococcal urethritis, condyloma acuminata, chlamydia trachomatis, chancroid, genital herpes, trichomoniasis, AIDS, cervical cancer, and so on.

[0003] Cervical cancer is the fourth most common cancer in terms of both incidence and mortality among women, second only to breast cancer, lung cancer, and colorectal cancer. And 99.8% of cervical cancers are caused by HPV infection. However, being infected with HPV does not mean having cervical cancer. It takes 5 to 10 years or even longer of continuous infection for it to possibly develop into cervical cancer. Moreover, male HPV infection may also cause skin irritation, such as condyloma acuminata, with discomfort symptoms such as skin epidermal ulceration and exudation, accompanied by itching, pain, and other reactions; it may also cause cross-infection, affecting sperm quality and reducing fertility. Therefore, it is particularly important to conduct HPV screening as early as possible. However, in the actual medical treatment process, most women do not have a very good experience in having HPV screening in the hospital. The hospital needs to use a professional speculum to insert the sampling consumables into the female vaginal cervix. This sampling method depends on the experience of clinical doctors and is invasive, which may cause micro-lesions or induce inflammatory reactions, resulting in a poor user experience. To a certain extent, this has led to a low penetration rate of HPV screening. The sampling for male HPV examination is mainly from the urethra. Currently, urethral sampling is mainly done with a cotton swab, which will cause certain discomfort. Moreover, there are certain defects in current urethral sampling, such as a small sampling volume and inability to sample multiple points. Currently, CN115363643B discloses a method for collecting and storing samples. It is necessary to pour PBS into the collection bag, put on the collection device, thoroughly scrub the urethral orifice, coronal sulcus, inside and outside the foreskin, scrotum, and the root of the penis, and then collect and send it to the laboratory for testing. Although this method reduces the discomfort of sampling and covers multiple-point sampling, the operation is still relatively cumbersome.

[0004] Gonorrhea is a classic sexually transmitted disease. For male patients, urethral specimens are generally collected, and pharyngeal specimens are added for those with a history of oral sex; for homosexuals, urethral, rectal, and pharyngeal specimens need to be collected. For female patients, cervical specimens are collected, and specimens are taken from the urethra, rectum, pharynx, greater vestibular gland, and paraurethral gland when necessary. The above collections need to be carried out with sampling swabs, which depend on clinical doctors and are invasive, and may cause micro-lesions or induce inflammatory reactions.

[0005] For the detection of reproductive tract-related diseases, a more accurate, comprehensive, gentle, and non-invasive sampling method is needed to improve the acceptability of patients and provide a more effective auxiliary means for the timely detection of reproductive tract diseases. Summary of the Invention

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art and provide the application of a condom used in sexual life in simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] In the first aspect, the present invention provides the application of a condom used in sexual life in simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts.

[0009] The present invention uses a condom to collect reproductive tract pathogens during sexual life. The collection method is more gentle, non-invasive, and the collected pathogens are more comprehensive, with high user acceptance. It can not only simultaneously collect pathogens in the male and female reproductive tracts, but also collect pathogen samples that cannot be collected by ordinary reproductive tract swabs or require multiple collections. By using a condom to collect reproductive tract pathogens during sexual life, the types and concentrations of pathogens collected can be increased, and the detection accuracy of pathogens can be improved.

[0010] As a preferred embodiment of the application of the present invention, the reproductive tract pathogens include human immunodeficiency virus (HIV), human papillomavirus (HPV), hepatitis B virus, hepatitis C virus, Treponema pallidum, Ureaplasma urealyticum, Chlamydia trachomatis in the reproductive tract, or Neisseria gonorrhoeae.

[0011] As a preferred embodiment of the application of the present invention, the reproductive tract pathogens include human papillomavirus (HPV) or Neisseria gonorrhoeae.

[0012] As a preferred embodiment of the application of the present invention, the reproductive tract pathogen is human papillomavirus (HPV).

[0013] As a preferred embodiment of the application of the present invention, the condom includes a condom body and a lubricant, and the lubricant includes a basic lubricating fluid and a cell capture component; the cell capture component is microspheres.

[0014] Microspheres are biodegradable microparticles with a porous structure and have a certain enrichment effect on cells. The present invention has found that adding a condom with a lubricant containing microspheres can more effectively enrich the cells in the reproductive tract, and thus collect more pathogens and increase the collection concentration of pathogens.

[0015] As a preferred embodiment of the application of the present invention, the material of the condom body is not limited, and is preferably natural rubber latex.

[0016] As a preferred embodiment of the application of the present invention, the cell capture component is poly(lactic acid) (PLLA) microspheres.

[0017] The condom of the present invention contains a lubricant of poly(lactic acid) (PLLA) microspheres. During sexual intercourse, human papillomavirus (HPV) is collected using this condom. It is found that the condom containing the lubricant of poly(lactic acid) (PLLA) microspheres has a significantly better effect of enriching cells compared with the condom containing an antibacterial gel lubricant and the condom containing only a hyaluronic acid lubricant, and can significantly increase the concentration of the collected human papillomavirus (HPV).

[0018] As a preferred embodiment of the application of the present invention, in the lubricant, the concentration of poly(lactic acid) (PLLA) microspheres is 50 - 100 mg / mL.

[0019] As a preferred embodiment of the application of the present invention, in the lubricant, the concentration of poly(lactic acid) (PLLA) microspheres is 50 mg / mL.

[0020] As a preferred embodiment of the application of the present invention, the main component of the basic lubricating fluid is hyaluronic acid.

[0021] As a preferred embodiment of the application of the present invention, the lubricant further includes a cell preservation solution.

[0022] The present invention finds through research that compared with the condom with a lubricant without a cell preservation solution, the condom with a lubricant containing a cell preservation solution has higher stability of the sample collected during sexual intercourse. The collected sample can be stored at room temperature for 1 - 2 months, and the concentration of the sample does not change significantly.

[0023] As a preferred embodiment of the application of the present invention, the cell preservation solution is composed of the following components in mass - volume percentage: sodium dihydrogen phosphate 2% - 4%, lactic acid 0.5% - 1.5%, lactobionic acid 3.5% - 3.6%, glutathione 0.05% - 0.1%, and water is added up to 100%.

[0024] The present invention uses sodium dihydrogen phosphate, lactic acid, lactobionic acid, glutathione and water as the cell preservation solution. The components used are all substances harmless to the human body, which can reduce the potential safety hazards to the human body. It can be directly used as a component of the condom lubricant, or directly coated on the outer membrane of the condom for the preservation of the collected cells. Commercially available cell preservation solutions usually contain components harmful or irritating to the human body and cannot directly enter the human body as an effective component of the condom lubricant.

[0025] Meanwhile, through research, the present invention discovers that the cell preservation solution of the present invention can achieve better preservation effects only within a specific range of the present invention.

[0026] As a preferred embodiment of the application of the present invention, the cell preservation solution is composed of the following components in mass-volume percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, and water is made up to 100%.

[0027] As a preferred embodiment of the application of the present invention, the pH of the cell preservation solution is 6 - 7.

[0028] As a preferred embodiment of the application of the present invention, the pH of the cell preservation solution is 6.5.

[0029] As a preferred embodiment of the application of the present invention, in the lubricant, the volume percentage of the cell preservation solution is 30% - 35%.

[0030] As a preferred embodiment of the application of the present invention, in the lubricant, the volume percentage of the cell preservation solution is 33.3%.

[0031] In a second aspect, the present invention provides a condom for simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts. The condom includes a condom body and a lubricant. The lubricant includes a basic lubricating fluid and a cell capture component; the cell capture component is microspheres.

[0032] As a preferred embodiment of the application of the present invention, the material of the condom body is not limited, and natural rubber latex is preferred.

[0033] As a preferred embodiment of the condom of the present invention, the cell capture component is poly(lactic acid) (PLLA) microspheres.

[0034] As a preferred embodiment of the condom of the present invention, in the lubricant, the concentration of poly(lactic acid) (PLLA) microspheres is 50 - 100 mg / mL.

[0035] As a preferred embodiment of the condom of the present invention, in the lubricant, the concentration of poly(lactic acid) (PLLA) microspheres is 50 mg / mL.

[0036] As a preferred embodiment of the condom of the present invention, the main component of the basic lubricating fluid is hyaluronic acid.

[0037] As a preferred embodiment of the condom of the present invention, the lubricant further includes a cell preservation solution.

[0038] As a preferred embodiment of the condom of the present invention, the cell preservation solution is composed of the following components in mass-volume percentage: sodium dihydrogen phosphate 2% - 4%, lactic acid 0.5% - 1.5%, lactobionic acid 3.5% - 3.6%, glutathione 0.05% - 0.1%, and water is made up to 100%.

[0039] As a preferred embodiment of the condom of the present invention, the cell preservation solution is composed of the following components in mass-volume percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, and water is made up to 100%.

[0040] As a preferred embodiment of the condom of the present invention, the pH of the cell preservation solution is 6 - 7.

[0041] As a preferred embodiment of the condom of the present invention, the pH of the cell preservation solution is 6.5.

[0042] As a preferred embodiment of the condom of the present invention, in the lubricant, the volume percentage of the cell preservation solution is 30% - 35%.

[0043] As a preferred embodiment of the condom of the present invention, in the lubricant, the volume percentage of the cell preservation solution is 33.3%.

[0044] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0045] The present invention uses a condom to collect genital tract pathogens during sexual intercourse. The collection method is milder, non-invasive, and can collect more comprehensive pathogens. The user acceptance is high. It can not only collect pathogens from both male and female genital tracts at the same time, but also collect pathogen samples that cannot be collected by ordinary genital tract swabs or need to be collected multiple times. Using a condom to collect genital tract pathogens during sexual intercourse can increase the types and concentrations of pathogen collection, and improve the detection accuracy of pathogens. At the same time, the present invention finds that using a condom containing microsphere lubricant can more effectively enrich genital tract cells, thereby increasing the concentration of pathogen collection and improving the detection accuracy of pathogens; using the cell preservation solution with specific components of the present invention can improve the preservation efficiency of the cells collected by the condom, extend the preservation time, improve the stability of the collected samples, and further improve the accuracy of sample detection. Specific Embodiments

[0046] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0047] For other materials, reagents, etc. used in the examples, unless otherwise specified, they can all be obtained from commercial channels. Example 1 Effect and Acceptance Test of Condom Sampling

[0048] 1. Collect human papillomavirus (HPV) from women using commercially available condoms (Guangzhou Wanfangjian Pharmaceutical Co., Ltd., registration certificate number: Guangdong Medical Device Registration No. 20182180489), and test the sampling effect of condoms.

[0049] (1) Sampling method

[0050] Use condoms for sampling and the HPV sampler for sampling as the reference sampling method.

[0051] (2) Test population

[0052] Select 20 gynecological outpatients, excluding those during menstruation, those who have received vaginal medication or vaginal douching within 3 days, and those who have had sexual intercourse within 24 hours before the test.

[0053] (3) Experimental method

[0054] ① Sample collection

[0055] 1) Divide the experimental population into two groups and number them, Group A and Group B;

[0056] 2) For the patients in Group A, first sample the vagina with an HPV sampler, and then sample with a condom (through sexual intercourse with a condom) after a 12-hour interval. The samples collected by different methods are immediately placed in a storage tube containing cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) for storage, and are sequentially recorded as samples 1 - 10;

[0057] 3) For the patients in Group B, first sample with a condom (through sexual intercourse with a condom), and then sample the vagina with an HPV sampler after a 12-hour interval. The samples collected by different methods are immediately placed in a storage tube containing cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) for storage, and are sequentially recorded as samples 11 - 20;

[0058] ② Sample elution and collection

[0059] Take out the samples of the two sampling methods from the two groups of Group A and Group B, and wash them thoroughly with cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) to ensure complete elution of the cells. The eluted samples are centrifuged and reserved for use.

[0060] ③ Sample detection

[0061] 1) Sample pretreatment

[0062] Use the "High-risk Human Papillomavirus Nucleic Acid (Typing) Detection Kit (PCR-Fluorescent Probe Method)" of Shengxiang Biotechnology Co., Ltd. and pretreat the samples strictly according to the instructions of the kit. The sample pretreatment solution can be used immediately or stored at 4°C for short-term storage and access.

[0063] 2) Add sample

[0064] Strictly follow the steps in the instructions for adding samples. Each sample to be tested should be added to the 4 reaction tubes of the same 8-tube strip and they should not be mixed up.

[0065] 3) PCR amplification

[0066] This experiment uses a PCR amplifier to amplify samples, and the sample tubes are placed and parameters are set according to the instructions.

[0067] 4) Result analysis

[0068] After the reaction is completed, the results are automatically saved and the CT value and results are recorded.

[0069] The test results of female HPV samples by different sampling methods are shown in Table 1.

[0070] Table 1 Detection results of female HPV samples by different sampling methods

[0071]

[0072]

[0073] Note: “+” indicates a positive test result, and “-” indicates a negative test result.

[0074] The results in Table 1 show that more types of HPV can be collected by using the condom sampling method. There were 7 HPV positive results in condom sampling, with a positive rate of 35%, and 5 HPV positive results in HPV sampler sampling, with a positive rate of 25%. Condoms do not have cross-reactions with HPV samples, and it is speculated that HPV sampler sampling may have missed detection. It is currently known that HPV sampler sampling is highly dependent on the doctor's experience, sampling techniques, etc., resulting in a missed detection rate of up to 50%. The CT value reflects the concentration of the nucleic acid detection content in the sample, generally referring to the number of cycles experienced when the fluorescent signal reaches the set value. A high CT value indicates a low virus concentration. In the experimental results, the CT value concentration of condom sampling in samples 1, 2, 6, 7, 11, 12, and 19 is higher than that of HPV sampler sampling, indicating that the cell collection effect of condom sampling is better and the HPV virus concentration is higher.

[0075] 2. Collect male human papillomavirus (HPV) using commercially available condoms (Guangzhou Wanfangjian Pharmaceutical Co., Ltd., registration number: Guangdong Medical Device Approval No. 20182180489), and test the sampling effect of condoms.

[0076] (1) Sampling method

[0077] Use condoms for sampling and cotton swabs for sampling as the reference sampling method.

[0078] (2) Test population

[0079] Select 10 confirmed male HPV patients (denoted as 1# - 10# respectively). Those who have had sexual intercourse within 24 hours before the test need to be excluded.

[0080] (3) Experimental method

[0081] ① Sample collection

[0082] Before sexual intercourse, first use a cotton swab to sample at the urethral orifice by yourself, and put the collected sample into a cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record number: Guangdong Sui Medical Device Preparation No. 20211302). 12 hours after sampling with the cotton swab, have sexual intercourse with a condom. After sexual intercourse, carefully remove the condom containing semen and place it in a condom collection box (containing cell preservation solution).

[0083] ② Sample elution and collection

[0084] Aspirate the sperm secretion inside the condom, and at the same time take out the cell preservation solution containing the sample collected by the cotton swab.

[0085] ③ Sample detection is the same as the experiment for female patients

[0086] The detection results of male HPV samples with different sampling methods are shown in Table 2 below.

[0087] Table 2 Detection results of male HPV samples with different sampling methods

[0088] Number CT value of cotton swab sampling CT value of condom sampling 1# 28.9 25.3 2# 36.3 28.5 3# 37.2 30.1 4# 36.7 30.3 5# 29.8 23.4 6# 35.3 29.0 7# 28.9 22.1 8# 35.4 26.8 9# 33.2 27.3 10# 30.1 25.4

[0089] The results in Table 2 show that the CT value concentration of condom sampling is higher than that of cotton swab sampling, indicating that condom sampling can collect more cells, the HPV virus concentration is higher, and the sampling effect is better.

[0090] 3. Acceptance of condom sampling

[0091] In order to collect more objective sampling information, a questionnaire survey was conducted on the subjects to investigate the acceptance of condom sampling by men and women in this example.

[0092] The questionnaire survey results of the acceptance of sampling methods are shown in Table 3 below.

[0093] Table 3 Results of the Acceptability Questionnaire on Sampling Methods

[0094]

[0095]

[0096] The results in Table 3 show that 90% of people are more accepting of the sampling method using condoms.

[0097] 4. Use the commercially available condoms (Guangzhou Wanfangjian Pharmaceutical Co., Ltd., registration certificate number: Guangdong Medical Device Registration No. 20182180489) to collect Neisseria gonorrhoeae, the pathogen of gonorrhea, and test the impact of condom sampling on gonorrhea test results.

[0098] For condom sampling, urethral / vaginal swab sampling is used as the reference sampling method.

[0099] Select 20 patients in the STD department (10 males and 10 females). For females, those in the menstrual period, those who have received vaginal medication or vaginal douching within 3 days, and those who have had sexual intercourse within 48 hours need to be excluded.

[0100] (1) Sample collection

[0101] ① Divide the experimental population into two groups and number them, Group A and Group B (each group consists of 5 male patients with a confirmed diagnosis and 5 female patients with a confirmed diagnosis).

[0102] ② For the female patients in Group A, first perform vaginal and cervical sampling; for male patients, first perform urethral sampling. After a 12-hour interval, perform condom sampling (by having sexual intercourse while wearing the condom of the present invention). The samples collected by different methods are immediately placed in a storage tube containing cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) for storage after collection.

[0103] ③ For the patients in Group B, first perform condom sampling (by having sexual intercourse while wearing the condom of the present invention). After an 8-hour interval, female patients perform vaginal and cervical sampling; male patients perform urethral sampling. The samples collected by different methods are immediately placed in a storage tube containing cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) for storage after collection.

[0104] (2) Sample elution and collection

[0105] Take out the samples of the two sampling methods from the two groups of Group A and Group B, and wash them thoroughly with cell preservation solution (Guangzhou Langkun Biotechnology Co., Ltd., record-filing number: Guangdong Medical Device Record-filing No. 20211302) to ensure complete elution of cells, and keep them at 4°C for standby.

[0106] (3) Sample testing

[0107] Select a Neisseria gonorrhoeae antigen detection kit and perform specific operations on the samples according to the instructions to count the results.

[0108] The detection results of different sampling methods for gonorrhea samples are shown in Table 4 below.

[0109] Table 4 Detection results of different sampling methods for gonorrhea samples

[0110]

[0111] Note: "+" indicates a positive test result, and "-" indicates a negative test result.

[0112] Table 4 shows that among the gonorrhea test results of 20 patient samples, 9 cases were positive for gonorrhea by condom sampling, with a positive rate of 45%, and 7 cases were positive by swab sampling, with a positive rate of 35%. From the test results, it can be seen that the condom sampling is 100% consistent with the hospital diagnosis results, while there is a missed detection situation with swab sampling. It shows that condom sampling has a better effect, no missed detection occurs, and it is non-invasive and more acceptable to users.

[0113] In summary, the method of sampling with a condom is feasible, and compared with the conventional swab, the sampling effect is better, more virus can be collected, the virus concentration is higher, and more virus types can be obtained. Sampling with a condom is completed during normal sexual intercourse, and there is no damage to cells or tissues caused by conventional swab sampling, which belongs to a non-invasive sampling method. At the same time, sampling with a condom during sexual intercourse has a larger sampling area, more complete sample collection, and can collect exfoliated cells and living cells at the same time, so more virus samples can be collected. At the same time, through a questionnaire survey, it is found that sampling with a condom during sexual intercourse has a high acceptance rate among users. It is a better method for collecting reproductive tract pathogens and also a better auxiliary means for detecting reproductive tract pathogens.

[0114] Example 2 Influence of porous materials on the condom sample collection effect

[0115] In this example, biodegradable microparticles of poly(lactic acid) (PLLA) microspheres with a porous structure and gels with a porous structure were selected to study the enrichment effect of condoms prepared with different porous materials on cells.

[0116] Mix the antibacterial gel (50 mg / mL) with hyaluronic acid to prepare a gel lubricant, and evenly apply the gel lubricant on the condom body. The condom containing the gel lubricant is Group A.

[0117] Mix poly(lactic acid) (PLLA) microspheres (50 mg / mL) with hyaluronic acid to prepare a PLLA microsphere lubricant, and evenly coat the lubricant on the condom body. The condom containing the PLLA microsphere lubricant is Group B;

[0118] Use hyaluronic acid as the lubricant, evenly apply the lubricant on the prepared condom body. The condom containing the hyaluronic acid lubricant is Group C;

[0119] Take the condoms of Group A, Group B, and Group C as objects, and test the effects of different condoms on cell enrichment respectively.

[0120] In addition, mix poly(lactic acid) (PLLA) microspheres at different concentrations (25 mg / mL, 50 mg / mL, 100 mg / mL, 150 mg / mL) with hyaluronic acid to prepare a PLLA microsphere lubricant, and evenly apply the lubricant on the prepared condom body to prepare PLLA microsphere condoms with different concentrations, denoted as Group a (25 mg / mL), Group b (50 mg / mL), Group c (100 mg / mL), and Group d (150 mg / mL).

[0121] In vitro simulation experiment

[0122] Cells: Use the human cervical cancer HPV18 positive cell line HeLa. Culture HeLa cells in a medium containing 10% fetal bovine serum at 37 °C in a 5% CO2 atmosphere, and collect HeLa cells to prepare a HeLa cell suspension.

[0123] Conduct simulation tests in vitro using the condoms of Group A, Group B, and Group C, and count the HeLa cells recovered by conventional methods.

[0124] Apply 5 mL of cell suspension containing an equal number of HeLa cells (1×10 6 cells) evenly on the outer membranes of the condoms of Group A, Group B, and Group C. After standing for 15 minutes, start elution. Use a cell preservation solution (the following components in mass-volume percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with water) to fully elute the cells. Collect the eluate for later use. Then measure the cell concentration by cell counting.

[0125] The cell capture situations of the condoms corresponding to different lubricants are shown in Table 5 below.

[0126] Table 5 Cell capture situations of the condoms corresponding to different lubricants

[0127]

[0128] Table 5 results show that the condom containing PLLA microsphere lubricant has the highest cell concentration, reaching 8×10 5 cells / mL, which is significantly higher than the cell concentrations of condoms containing gel lubricant and ordinary condoms, indicating that PLLA microspheres help condoms capture more cells.

[0129] Similarly, simulated tests were carried out in vitro using condoms of groups a, b, c, and d, and HeLa cells were recovered by conventional methods for counting. The method is the same as the above-mentioned experiments of groups A, B, and C in this example.

[0130] The effects of lubricants with different PLLA microsphere concentrations on cell capture are shown in Table 6 below.

[0131] Table 6 Effects of lubricants with different PLLA microsphere concentrations on cell capture

[0132]

[0133] Table 6 results show that the concentration of PLLA microspheres will significantly affect the cell concentration after treatment with PLLA microsphere condoms and the adsorption efficiency of condoms containing PLLA microsphere lubricant on cells. When the concentration of PLLA microspheres is too low, at 25 mg / mL, the cell concentration of the condom containing PLLA microsphere lubricant is only 2×10 4 cells / mL. After the concentration of PLLA microspheres increases, the cell concentration gradually increases. However, when the concentration of PLLA microspheres increases to a certain extent, further increase will lead to a decrease in cell concentration. This shows that the concentration of PLLA microspheres can have a better effect of adsorbing / enriching cells within a specific range. When the concentration of PLLA microspheres is in the range of 50 - 100 mg / mL, the cell concentration is relatively high, with good cell adsorption efficiency and good cell enrichment effect.

[0134] Example 3 Influence of cell preservation solution on sample collection effect

[0135] 1. Influence of the concentration of different components of the cell preservation solution on cell preservation effect.

[0136] Sodium dihydrogen phosphate, lactic acid, lactobionic acid, and glutathione were dissolved in deionized water to prepare a cell preservation solution.

[0137] Cells: Human cervical cancer HPV18 positive cell line HeLa. HeLa cells were cultured using 10% fetal bovine serum under the atmosphere of 37°C and 5% CO2, and HeLa cell suspension was prepared by collecting HeLa cells.

[0138] Configuration of cell preservation solution: Prepare 20 mL of cell preservation solution with the same volume for each, numbered as groups A - D:

[0139] Group A

[0140] The following components by mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 2%, lactic acid 0.5%, lactobionic acid 3.5%, glutathione 0.05%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0141] Group B

[0142] The following components by mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0143] Group C

[0144] The following components by mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 1%, lactic acid 0.5%, lactobionic acid 3.4%, glutathione 0.03%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0145] Group D

[0146] The following components by mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 5%, lactic acid 2%, lactobionic acid 3.7%, glutathione 0.15%, adjust the pH to 6.5, and make up to 100% with deionized water.

[0147] Add HeLa cell suspensions containing equal amounts of HeLa cells to condoms containing the cell preservation solutions of each group respectively for cell preservation tests. Preserve HeLa cells at room temperature for 5 days, 2 weeks, 1 month, and 2 months. After different preservation times, centrifuge the samples containing the HeLa cell preservation solution at 10,000 rpm for 1 minute to remove the supernatant. Recover the HeLa cells, select a high-risk human papillomavirus nucleic acid (genotyping) detection kit (PCR-fluorescent probe method), strictly perform sample pretreatment according to the kit instructions, and strictly add samples according to the steps in the instructions. Add each test sample to 4 reaction tubes in the same octuplet tube respectively without confusion. Use a PCR amplifier for amplification reactions, and place the sample tubes and set parameters according to the instructions. After the reaction ends, the results are automatically saved, and the CT values and results are recorded.

[0148] The effects of different concentrations of each component in the cell preservation solution on the cell preservation effect are shown in Table 7 below,

[0149] Table 7 Effects of Different Concentrations of Each Component in the Cell Preservation Solution on the Cell Preservation Effect

[0150]

[0151] Note: NA indicates no result.

[0152] The results in Table 7 show that the CT values detected for the cells preserved in the cell preservation solutions of groups A and B are not significantly different, indicating that the cells are stable. That is, the cell preservation solutions of groups A and B are more suitable for cell preservation.

[0153] The components of the cell preservation solution selected in the present invention: sodium dihydrogen phosphate, lactic acid, lactobionic acid, and glutathione are all substances harmless to the human body, which can reduce potential safety hazards to the human body. It can be directly used as a component of condom lubricant for cell preservation.

[0154] Commercially available cell preservation solutions usually contain one or more of the following components, including methanol, Tween, glacial acetic acid, proclin950, tris(hydroxymethyl)aminomethane hydrochloride, dodecyltrimethylammonium chloride, etc. As is well known, these substances have more or less certain effects / toxicity on the human body and cannot directly enter the human body.

[0155] Methanol and glacial acetic acid: They fix cells and the cell nucleus is well preserved. However, methanol is a transparent, colorless, and toxic volatile liquid that can enter the human body through the respiratory tract, skin, and digestive tract, causing poisoning. Glacial acetic acid has high toxicity, and inhalation or contact with glacial acetic acid can cause respiratory infections, asthma, pulmonary edema, etc., and can even endanger life in severe cases.

[0156] Tween: As a surfactant, more and more studies have shown this year that Tween also has certain toxicity to the human body.

[0157] Tris(hydroxymethyl)aminomethane hydrochloride: It is usually used in the Tris-HCL buffer system and has low toxicity, but it can cause cancer and should not be directly contacted with the skin.

[0158] Dodecyltrimethylammonium chloride: As a cationic surfactant, it has relatively high safety for the human body and the environment under normal use conditions. However, due to its certain toxicity, it needs to be used and stored correctly. Long-term contact or large intake may have adverse effects on human health, such as irritation, allergy, etc. Therefore, correct protective measures should be observed during use to minimize direct contact as much as possible. Thus, it cannot be directly used as a component of condom lubricant for cell preservation during sample collection.

[0159] 2. Influence of different components of the cell preservation solution on cell preservation effect.

[0160] Cells: Human cervical cancer HPV18 positive cell line HeLa. HeLa cells were cultured in an atmosphere of 10% fetal bovine serum at 37°C and 5% CO2, and HeLa cell suspension was prepared by collecting HeLa cells.

[0161] Preparation of the cell preservation solution: Prepare 20 mL of cell preservation solutions with the same volume for each group, numbered a - d. Group a

[0162] The following components in terms of mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0163] Group b

[0164] The following components in terms of mass (g), volume (mL), and percentage: sodium bicarbonate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0165] Group c

[0166] The following components in terms of mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, trehalose 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0167] Group d

[0168] The following components in terms of mass (g), volume (mL), and percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, vitamin C 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water;

[0169] Commercially available cell preservation solution: Cell preservation solution PreservCyt Solution, PreservCyt (registered trademark) (Hologic Inc.), and the detailed composition is not disclosed.

[0170] Add HeLa cell suspensions containing an equal number of HeLa cells (1×10 6 cells) to the cell preservation solutions of groups a - d (12 mL) and the commercially available cell preservation solution (12 mL) respectively. Store these cells at temperature conditions of 1°C and 31°C for 2 weeks, 1 month, and 2 months respectively. After different storage times, centrifuge the samples containing the HeLa cell preservation solution at 10,000 rpm for 1 minute to remove the supernatant. Recover the HeLa cells, and extract DNA using the "High-risk human papillomavirus nucleic acid (genotyping) detection kit (PCR-fluorescent probe method)" of Shengxiang Biotech Co., Ltd. (The specific operation is carried out according to the kit instructions), perform PCR amplification on the extracted DNA, and record the CT values and results.

[0171] The effects of different components of the cell preservation solution on cell preservation are shown in Table 8 below,

[0172] Table 8 Effects of different components of the cell preservation solution on cell preservation

[0173]

[0174] Note: NA indicates no result.

[0175] Table 8 results show that when the cell preservation solution of group a was used to preserve cells for 2 months, the change in CT value of the cells was small and not significant. However, when the commercially available cell preservation solution was used to preserve cells for 2 months, the CT value of the cells increased significantly, indicating that the cell preservation solution of group a (the specific components of the present invention) is more stable for cell preservation. When the cell preservation solutions of groups b, c, and d were used to preserve cells for 2 months, the CT value of the cells increased significantly or could not be detected, indicating that the cell preservation solutions of groups b, c, and d have poor stability for cell preservation, indicating that the cell preservation solution composed of the components of group a is more stable.

[0176] 3. Test on the stability of the cell preservation solution

[0177] Cell preservation solution: the following components in mass (g) per volume (mL) percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, adjust the pH to 6.5, and make up to 100% with deionized water.

[0178] Mix the cell preservation solution (500 mL) with hyaluronic acid (1000 mL) to prepare a lubricant, and coat it on the condom body to explore the preservation stability of cell DNA in the condom lubricant containing the cell preservation solution. The condom with hyaluronic acid as the lubricant was used as a control.

[0179] During sexual intercourse, cervical cells of 12 subjects who have been determined to be HPV positive were collected using condoms. All 12 subjects needed to be sampled twice (the interval between the two samplings was 12 hours). Among the 12 subjects, 8 subjects used condoms containing the cell preservation solution and were respectively denoted as A - H, and the other 4 subjects used condoms without the cell preservation solution and were respectively denoted as a - d. The CT value detected in the first sampling of the 12 subjects was recorded as the starting value. The samples of the second sampling of subjects A - D and a - d were placed at room temperature for 1 week and then the CT value was detected. The samples of the second sampling of subjects E - H were placed at room temperature for 2 weeks and then the CT value was detected.

[0180] At the predetermined storage time, after eluting the outer surface of the condom at each time point with 2 mL of the cell preservation solution, centrifuge at 10,000 rpm for 1 minute to remove the supernatant. Recover the cells, and extract DNA using the "High - risk Human Papillomavirus Nucleic Acid (Genotyping) Detection Kit (PCR - Fluorescent Probe Method)" of Shengxiang Biotech Co., Ltd. (the specific operation is carried out according to the kit instructions). Perform PCR amplification on the extracted DNA, and record the CT value and results.

[0181] The influence of the cell preservation solution on the samples is shown in Table 9 below.

[0182] Table 9 Influence of the cell preservation solution on the samples

[0183]

[0184] Note: NA means no result, “ / ” means not detected.

[0185] The results in Table 9 show that the DNA of the samples collected with condoms containing cell preservation solution remained basically stable after two weeks, but the DNA of the condom samples without cell preservation solution could not be detected within one week, which may be due to the lysis of cells and degradation of nucleic acid.

[0186] Example 4: Test on the influence of condom sampling on sample test results

[0187] Cell preservation solution: the following components in mass volume percentage: 4% sodium dihydrogen phosphate, 1.5% lactic acid, 3.6% lactobionic acid, 0.1% glutathione, supplemented to 100% with deionized water, and adjusted to pH 6.5.

[0188] Cell preservation solution (500 mL), polylactic acid (PLLA) microspheres (50 mg / mL) and hyaluronic acid (1000 mL) were mixed to prepare a lubricant containing cell preservation solution and polylactic acid (PLLA) microspheres, and the mixture was coated on the condom body to prepare a condom containing cell preservation solution and polylactic acid (PLLA) microspheres in the lubricant. An in vitro simulation experiment was conducted to study the effect of condom sampling on the HPV detection results in the samples.

[0189] 1. Experimental methods

[0190] 60 condoms of this embodiment were selected and divided into 2 groups, 30 each. Group A: The outer wall of the condom was evenly smeared with semen containing HPV recombinant plasmid, and Group B: The outer wall of the condom was evenly smeared with the same amount of semen as Group A (without HPV recombinant plasmid); after smearing, it was allowed to stand for 15 minutes, and 2mL of cell preservation solution was used to elute the outer side of the condoms of the two groups AB, and all the liquids after washing were collected for centrifugation. The "High-risk Human Papillomavirus Nucleic Acid (Typing) Detection Kit (PCR-Fluorescent Probe Method)" of Shengxiang Biotechnology Co., Ltd. was selected, and the sample was added strictly according to the steps in the instructions. Each sample to be tested was added to the 4 reaction tubes of the same eight-tube, and they could not be confused. The PCR amplifier used was placed and the parameters were set according to the instructions. After the reaction was completed, the results were automatically saved, and the CT value and results were recorded.

[0191] The effect of condoms on HPV test results is shown in Table 10 below.

[0192] Table 10 Effect of condoms on HPV test results

[0193]

[0194]

[0195] Note: “+” indicates that the test result is positive; “-” indicates that the test result is negative and the CT value has no result.

[0196] The results in Table 10 show that none of the samples that did not contain HPV plasmids had HPV-positive results, proving that the condom itself has no effect on HPV detection.

[0197] Condoms are commonly used tools during sexual intercourse, which can effectively prevent the spread of diseases in the reproductive tract. At present, cervical / urethral sampling does not really touch the cervix, especially the cervical lesion site, and the sampling is incomplete and cannot reflect the full picture of the disease. At the same time, cotton swab sampling has strong adsorption, resulting in poor elution effect or detection results. The present invention uses condoms for sampling, which reduces the discomfort caused by cervical brush / urethral sampling, has a lower sense of rejection, better sampling effect, and more comprehensive sampling sites. It can be seen from various embodiments that condom sampling has a higher accuracy in result detection.

[0198] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. Application of a condom after sexual intercourse in the simultaneous collection and / or detection of genital tract pathogens in male and female; The condom includes a condom body and a lubricant, and the lubricant includes a basic lubricant and a cell capture component; the cell capture component is porous polylactic acid microspheres, and the concentration of the porous polylactic acid microspheres is 50-100 mg / mL; The lubricant further includes a cell preservation solution, and the cell preservation solution is composed of components in the following mass-volume percentages: Sodium dihydrogen phosphate 2%-4%, lactic acid 0.5%-1.5%, lactobionic acid 3.5%-3.6%, glutathione 0.05%-0.1%, water is added to make up 100%; The genital tract pathogens include human papillomavirus or Neisseria gonorrhoeae.

2. The application according to claim 1, characterized in that, The genital tract pathogen is human papillomavirus.

3. A condom for simultaneously collecting and / or detecting pathogens in the male and female reproductive tracts, the condom comprising a condom body and a lubricant, characterized in that, The lubricant includes a basic lubricant and a cell capture component; the cell capture component is porous polylactic acid microspheres, and the concentration of the porous polylactic acid microspheres is 50-100 mg / mL; The lubricant further includes a cell preservation solution, and the cell preservation solution is composed of the following components in mass-volume percentage: sodium dihydrogen phosphate 2%-4%, lactic acid 0.5%-1.5%, lactobionic acid 3.5%-3.6%, glutathione 0.05%-0.1%, water is added to make up 100%.

4. The condom according to claim 3, wherein The cell preservation solution is composed of the following components in mass-volume percentage: sodium dihydrogen phosphate 4%, lactic acid 1.5%, lactobionic acid 3.6%, glutathione 0.1%, water is added to make up 100%.

5. The condom according to claim 3 or 4, characterized in that, The pH of the cell preservation solution is 6-7.

6. The condom according to claim 5, wherein the pH of the cell preservation solution is 6.5.

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