CN101271105B - Spermatozoon activity fast detecting reagent kit and its preparation and use method - Google Patents

Spermatozoon activity fast detecting reagent kit and its preparation and use method Download PDF

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Publication number
CN101271105B
CN101271105B CN 200810112085 CN200810112085A CN101271105B CN 101271105 B CN101271105 B CN 101271105B CN 200810112085 CN200810112085 CN 200810112085 CN 200810112085 A CN200810112085 A CN 200810112085A CN 101271105 B CN101271105 B CN 101271105B
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sample
swimming pool
test
millimeters
sperm
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CN 200810112085
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CN101271105A (en
Inventor
周明非
刘淑红
赵永岐
王毅
朱望
李娅
赵秀丽
邢玉斌
魏华
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WANGSHENG WEIYE TECH DEVELOPMENT Co Ltd BEIJING
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WANGSHENG WEIYE TECH DEVELOPMENT Co Ltd BEIJING
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Abstract

The invention discloses a sperm activity rapid testing kit, a preparation method and a using method thereof, and the kit comprises a sample collection part and a test part, wherein, the sample collection part comprises a sample collector and a sample swimming pool; the shape of the sample collector is a circular small bowl, the bottom part thereof is provided with a concave cylindrical small hole; the sample swimming pool is a cylindrical container, in which sperm swimming liquid is contained, both ends of the container are provided with sealing films for sealing; the outer diameter of the sample swimming pool is smaller than the inner diameter of the small hole of the bottom part of the sample collector, then the sample swimming pool can be inserted in the small hole of the bottom part of the sample collector; the test part comprises a test plate, dyeing liquid, lotion and a sampling dropper; the test plate comprises an outer shell and a substrate, the center of the outer shell is provided with two small holes of a test hole and a control hole, a water absorbent paper, a filter membrane and a standard color tag are arranged on the substrate, and the color of the standard color tag is the color after dyeing and washing of 20 million/ml of the sperm sample with the activity which is added in the test hole of the test plate.

Description

Spermatozoon activity fast detecting reagent kit and preparation thereof and method of application
Technical field
The present invention relates to a kind of in-vitro diagnosis class medicine equipment, specifically is a kind of quick detection kit that is applicable to that mankind spermatozoon is active.
Background technology
The fertility expert report, (no contraceptives) can not be conceived in 1 year for about in the world 15% Mr. and Mrs, and 40% in these cases are because male factor causes, also have male sex expert's report, and the incidence of disease of male sterility is approximately 6-12%.Above-mentioned report, the crowd that male sterility is described is a sizable crowd.
The main method of current diagnosis male sterility is the microscopically diagnosis of seminal fluid, and promptly sperm morphological detects, and this also is the goldstandard of male sterility diagnosis.But there are many shortcomings in this method, such as: because Chinese traditional concept is shyly gone the disease of hospital therapy male sterility aspect, thereby has been incured loss through delay treatment.Even gone to hospital, owing to the condition of getting sperm is limited, receive the restriction of environment, method, time, be difficult for obtaining sample.It is more loaded down with trivial details to go to a hospital to see a doctor, must be through a series of programs such as registering, prescribe.
Summary of the invention
The objective of the invention is in order to overcome the shortcoming of prior art, provide a kind of and can operate at home, can detect the whether qualified spermatozoon activity fast detecting reagent kit of activated sperm quantity fast, easy to use, low-cost.
Second purpose of the present invention provides a kind of preparation and method of application of spermatozoon activity fast detecting reagent kit.
For achieving the above object, the present invention includes sample collecting part and test portion, sample collecting partly comprises sample collecting device and sample swimming pool; The column type aperture that rounded little bowl-shape of sample collecting device, bottom have depression to go down; The sample swimming pool is cylindrical container, in be loaded with sperm swimming liquid, the container two ends are all sealed film and are sealed; The external diameter of sample swimming pool is slightly smaller than the internal diameter of the aperture of sample collecting device bottom, and the sample swimming pool can be inserted in the aperture of sample collecting device bottom; Test portion comprises breadboard, dyeing liquor, washing lotion, sampling dropper; Breadboard comprises shell, substrate; Shell and substrate are that rectangular PVC plastics are processed, and there are test hole and two apertures of control wells in shell central authorities, and thieving paper, filtering membrane and standard colour code are arranged on the said substrate; Thieving paper and filtering membrane are positioned at the substrate top, the test hole below of shell; Thieving paper is positioned at the filtering membrane below; The standard colors mark is in the substrate top, below the control wells; Standard colors target color is the test hole process dyeing of 2,000 ten thousand/milliliter activated sample of sperm adding breadboards, the color after the washing;
A kind of preparation method of spermatozoon activity fast detecting reagent kit comprises the steps:
Preparation 0.1-0.5% methylene blue solution: add phosphate buffer to methylene blue and fully dissolve, filter rearmounted room temperature preservation;
The preparation standard colour code: use 2,000 ten thousand/milliliter activated sperm samples to be master sample, the test hole that adds breadboard according to the color of experimental result, is prepared the standard colour code through after dyeing, washing, and places the control wells below;
Filtering membrane and thieving paper are placed the substrate top successively, the test hole below.
Advantage of the present invention: whether the quantity that can detect activated sperm rapidly and accurately is qualified; Thereby diagnose out the male sex whether can procreate, save a lot of times, use simple than traditional in the past diagnostic method; Purchase gets final product in pharmacy; Do not spend hospital admission, make the patient not have shy sense, make the man and wife can know the potential fertility Issue of oneself in advance.
Description of drawings
Below in conjunction with accompanying drawing to further detailed explanation of the present invention.
Fig. 1 is the sectional view of sample collecting device of the present invention;
Fig. 2 is the structural representation of sample swimming pool of the present invention;
Fig. 3 is the structural representation of breadboard of the present invention.
1. cylindrical aperture 2. sperms swimming liquid 3. seal film 4. shells 5. substrates 6. test holes 7. control wells 8. filtering membranes 9. thieving papers 10. standard colour codes among the figure
Embodiment
Can know that by figure the present invention includes sample collecting part and test portion, sample collecting partly comprises sample collecting device and sample swimming pool; The column type aperture 1 that rounded little bowl-shape of sample collecting device, bottom have depression to go down; The sample swimming pool is cylindrical container, in be loaded with sperm swimming liquid 2, the container two ends are all sealed film 3 and are sealed; The external diameter of sample swimming pool is slightly smaller than the internal diameter of the cylindrical aperture 1 of sample collecting device bottom, and the sample swimming pool can be inserted in the cylindrical aperture 1 of sample collecting device bottom; Test portion comprises breadboard, dyeing liquor, washing lotion, sampling dropper.Described sample collecting device is 50~60 millimeters of the diameters processed by the PVC plastics, high 25~35 millimeters circular little bowl, and the diameter of the column type aperture 1 that the bottom notch of little bowl is gone down is 10~12 millimeters, and is high 15~20 millimeters.Described sample swimming pool is that the clear PET plastics are processed, and external diameter is 9~11 millimeters, and internal diameter is 7~9 millimeters, and height is 60~80 millimeters.Breadboard comprises shell 4, substrate 5; Shell 4 is that rectangular PVC plastics are processed with substrate 5; There are test hole 6 and 7 two apertures of control wells in shell central authorities; Thieving paper 9, filtering membrane 8 and standard colour code 10 are arranged on the said substrate, and thieving paper 9 is positioned at substrate 5 tops, test hole 6 belows of shell 4 with filtering membrane 8; Thieving paper 9 is positioned at filtering membrane 8 belows; The standard colors mark is in substrate 5 tops, control wells 7 belows.The color of standard colour code 10 is the test hole process dyeing of 2,000 ten thousand/milliliter activated sample of sperm adding breadboards, the color after the washing.Described dyeing liquor is the 0.1-0.5% methylene blue solution.Described washing lotion is a physiological saline, also can be purified water and the destainer that contains acidic materials such as low-concentration acetic acid.Described sperm swimming liquid 2 is acrylic amide chlorination sodium solution.
The method for preparing spermatozoon activity fast detecting reagent kit is an example with 0.1% methylene blue solution coloring agent, comprises the steps:
Preparation concentration is 0.1% methylene blue solution: precision takes by weighing 105 ℃ of biological stain serge blue 100mg that are dried to constant weight, puts in the 1000ml measuring bottle, adds phosphate buffer to scale, shakes up, and filters, and filters rearmounted room temperature preservation;
The preparation standard colour code: use 2,000 ten thousand/milliliter activated sperm samples to be master sample, the test hole that adds breadboard according to the color of experimental result, is prepared the standard colour code through after dyeing, washing, and places the control wells below;
Filtering membrane and thieving paper are placed the substrate top successively, the test hole below.
Use the method for spermatozoon activity fast detecting reagent kit: comprising following steps:
Collect semen sample in sample collecting device;
Treat that semen sample liquefaction back flows in the aperture of sample collecting device bottom, the general time is 30 to assign to 60 minutes;
Open the film that seals of sample swimming pool; Because of swimming liquid is gel; Have surface tension, seal that swimming liquid can not flow out at once behind the film from the sample swimming pool, the sample swimming pool is inserted the aperture of sample collecting device bottom so open; Activated sperm can upwards move about along swimming liquid, and accumulates in the upper strata of swimming liquid;
Add the swimming liquid on a sample swimming pool upper strata with the sampling dropper, join in the test hole of breadboard, let test hole inner filtration film fully invade wet 1 minute;
Add dyeing liquor, add two dyeing liquors, let test hole inner filtration film fully invade wet 1 minute to test hole;
Add washing lotion, add two washing lotions, let test hole inner filtration film fully invade wet 1 minute to test hole;
The change color in viewing test hole:
The test hole blueness is darker than control wells, shows spermatozoon activity >=2,000 ten thousand/milliliter
The test hole blueness is blue more shallow than control wells, shows spermatozoon activity<2,000 ten thousand/milliliter
To in 3 minutes, read the result, inaccurate above reading the possibility of result after 3 minutes;
Test for the second time: per twice test is at interval more than the week.

Claims (4)

1. a spermatozoon activity fast detecting reagent kit comprises sample collecting part and test portion, and it is characterized in that: sample collecting partly comprises sample collecting device and sample swimming pool; The column type aperture that rounded little bowl-shape of sample collecting device, bottom have depression to go down; The sample swimming pool is cylindrical container, in be loaded with sperm swimming liquid, sperm swimming liquid is acrylic amide chlorination sodium solution, the container two ends are all sealed film and are sealed; The external diameter of sample swimming pool is slightly smaller than the internal diameter of the aperture of sample collecting device bottom, and the sample swimming pool can be inserted in the aperture of sample collecting device bottom; Test portion comprises breadboard, dyeing liquor, washing lotion, sampling dropper; Dyeing liquor is that 0.1-0.5% methylene blue solution, washing lotion are physiological saline; Breadboard comprises shell, substrate; Shell and substrate are that rectangular PVC plastics are processed, and there are test hole and two apertures of control wells in shell central authorities, and thieving paper, filtering membrane and standard colour code are arranged on the said substrate; Thieving paper and filtering membrane are positioned at the substrate top, the test hole below of shell; Thieving paper is positioned at the filtering membrane below; The standard colors mark is in the substrate top, below the control wells; Standard colors target color is the test hole process dyeing of 2,000 ten thousand/milliliter activated sample of sperm adding breadboards, the color after the washing.
2. a kind of spermatozoon activity fast detecting reagent kit according to claim 1; It is characterized in that: described sample collecting device is 50~60 millimeters of the diameters processed by the PVC plastics; High 25~35 millimeters circular little bowl; The diameter of the column type aperture that the bottom notch of little bowl is gone down is 10~12 millimeters, and is high 15~20 millimeters.
3. a kind of spermatozoon activity fast detecting reagent kit according to claim 1 is characterized in that: described sample swimming pool is that the clear PET plastics are processed, and external diameter is 9~11 millimeters, and internal diameter is 7~9 millimeters, and height is 60~80 millimeters.
4. the method for preparing each described a kind of spermatozoon activity fast detecting reagent kit of claim 1~3, comprising following steps:
Preparation 0.1-0.5% methylene blue solution: add phosphate buffer to methylene blue and fully dissolve, filter rearmounted room temperature preservation;
The preparation standard colour code: use 2,000 ten thousand/milliliter activated sperm samples to be master sample, the test hole that adds breadboard according to the color of experimental result, is prepared the standard colour code through after dyeing, washing, and places the control wells below;
Filtering membrane and thieving paper are placed the substrate top successively, the test hole below.
CN 200810112085 2008-05-21 2008-05-21 Spermatozoon activity fast detecting reagent kit and its preparation and use method Active CN101271105B (en)

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Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8932543B2 (en) * 2011-09-21 2015-01-13 Sakura Finetek U.S.A., Inc. Automated staining system and reaction chamber
CN102352305B (en) * 2011-09-27 2013-04-03 广州市第一人民医院 Sperm optimizing microflow control device and sperm optimizing method
CN104479994B (en) * 2014-12-31 2016-06-01 北京望升伟业科技发展有限公司 Family expenses motility of sperm test card
CN106645672A (en) * 2017-02-20 2017-05-10 刘晓强 Semen analysis system capable of accurately analyzing quality of semen
FR3081321B1 (en) * 2018-05-24 2020-06-12 Imv Technologies ANIMAL SEED COLLECTION DEVICE

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004053465A2 (en) * 2002-12-09 2004-06-24 Advanced Fluidix Laboratories, Llc Male fertility assay method and device
CN201203612Y (en) * 2008-05-21 2009-03-04 北京望升伟业科技发展有限公司 Kit for rapidly detecting spermatozoa activity
CN201392428Y (en) * 2009-05-07 2010-01-27 李西营 Novel sperm detection glass slide
CN101726578A (en) * 2008-10-27 2010-06-09 深圳科瑞克医疗器械有限公司 Microfluidic biological chip sperm quality analyser

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004053465A2 (en) * 2002-12-09 2004-06-24 Advanced Fluidix Laboratories, Llc Male fertility assay method and device
CN201203612Y (en) * 2008-05-21 2009-03-04 北京望升伟业科技发展有限公司 Kit for rapidly detecting spermatozoa activity
CN101726578A (en) * 2008-10-27 2010-06-09 深圳科瑞克医疗器械有限公司 Microfluidic biological chip sperm quality analyser
CN201392428Y (en) * 2009-05-07 2010-01-27 李西营 Novel sperm detection glass slide

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