CN105806744A - Method and device for rapidly, easily and conveniently judging viscosity of seminal fluid - Google Patents
Method and device for rapidly, easily and conveniently judging viscosity of seminal fluid Download PDFInfo
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- CN105806744A CN105806744A CN201610143415.3A CN201610143415A CN105806744A CN 105806744 A CN105806744 A CN 105806744A CN 201610143415 A CN201610143415 A CN 201610143415A CN 105806744 A CN105806744 A CN 105806744A
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- G01N11/00—Investigating flow properties of materials, e.g. viscosity, plasticity; Analysing materials by determining flow properties
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B2010/0003—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements including means for analysis by an unskilled person
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Abstract
The invention relates to a method and device for rapidly, easily and conveniently judging and analyzing whether the viscosity of the male seminal fluid is normal or not.A glass slide with a certain rough surface is adopted, the seminal fluid sample naturally flows under the gravity effect starting from the fixed position concentration of the glass slide with a certain inclination angle, whether the front flow end of the seminal fluid exceeds a marker line or not is checked within the required time, and whether the viscosity of the seminal fluid sample reaches the minimum standard of male fertility or not is judged based on the result.The invention further provides the device for executing the method and a kit.It is proved through comparative studies that the judgment result of the method and the judgment results of a pipette method and glass rod method of standard methods recommended by WHO have no statistical difference, and it is shown that the method can be used for rapidly checking whether the viscosity of the seminal fluid sample is normal or not and assist in evaluation of the activity of male sperms.More importantly, by means of the method, an examinee can perform operation by himself at a table, evaluation of the viscosity of the seminal fluid is completed, and reference is provided for further screening of infertility.
Description
Technical field
The present invention relates to and analyze the fast and convenient decision method that whether normal semen viscosity is.Adopt the microscope slide of certain rough surface, semen sample is made to start to lean on trickling naturally action of gravity from the fixed position concentration of the microscope slide at certain angle of inclination, check whether seminal fluid trickling front end exceedes markings at the appointed time, judge whether the viscosity of semen sample reaches the minimum standards of male fertility with this result.The qualitative judgement result of this method contributes to analyzing the reason of male sterility.
Background technology
Male infertility is one of sixty-four dollar question of human reproduction, especially the modal clinical problem of andrology and serious challenge.Owing to mankind spermatozoon quality declines with quantity since nearly 30～50 years, the trend that carcinoma of testis, testicular descent are bad, the sickness rate of hypospadias and infertility constantly raises, male infertility attention.Although in recent years diagnosing and treating of male infertility being achieved obvious progress, but male infertility is still that the extremely difficult problem that andrology clinician faces up to now, its reason is that people are to male genetic, particularly spermatogenetic understanding is far unclear, infertility is not independent sick kind simultaneously, it is subject to the impact of the many factors such as environment, heredity, in male infertility, the actual cause of disease of patient more than 50% still belongs to unknown, and this just necessarily increases the difficulty in clinical position.
The factor causing Mr. and Mrs sterile is numerous, complicated, relates to men and women's reproduction and holistic health, and the clear and definite specific causative agent of any sick kind is not only selection, and rationally and targetedly therapeutic scheme institute is required, is also the objective basis of judging prognosis.World Health Organization (WHO) (WHO) finds in 25 countries 9000 multicenter study to infertile couples, and the male infertility of 50% is that idiopathic oligoastheno is abnormal, wherein the human spermatogoa vigor not enough (azoospermia) of about 40%.Azoospermia is likely to and the nonspecific infection (such as coli-infection) of varicocele, genito-urinary system, the factors such as some metabolism medicine, antimalarial, estrogen, nitromin relevant, therefore with environmental pollution and drug dependence, mankind spermatozoon hypodynamic problem alive will tend to increasing the weight of.
Sperm must be travelling in the liquid environment that seminal fluid provides, and leaves seminal fluid and cannot move, and therefore the viscosity of seminal fluid has a strong impact on the travelling of sperm, and then affects motility of sperm and become pregnant.If the seminal fluid viscosity of post liquefaction is too high, can cause sterile to a certain extent, it is estimated that, in male sterility, nearly about 9% causes owing to human semen viscosity is too high.
World Health Organization (WHO) 2011 human seminal fluid checks that the human semen viscosity inspection method recommended with treatment of laboratory handbook has two kinds, is dropper method and Glass rod fiber elongation method respectively, judges that whether normal the viscosity of semen sample is by observing the length of wire drawing.When length of string is less than 2 centimetres, namely think that semen sample viscosity is normal, when length of string is more than 2 centimetres, namely think that semen sample viscosity is extremely (abnormal).
Although the method that WHO recommends is easy, but need more professional equipment, the such as disposable plastic pipette in wide-mouth footpath (diameter is about 1.5mm), or Glass rod, and pop-off after wire drawing meeting formation at short notice, accurately measure its length relatively difficult, and cannot complete voluntarily on limit on record, it is therefore desirable in specialized laboratory, completed by veteran technician.But semen is because relating to privacy problem, suspecting that the tender subject relating to oneself male ability is often taken great pains to guard a secret by sperm concentration and the problematic male of vigor, the detection preferring to need third-party involvement is reluctant in pole.
Currently invention addresses the analysis of semen viscosity, disclose the decision method that whether normal a kind of semen viscosity completed fast and convenient, self-service is, male is made very privacy to complete the analysis that whether normal human semen viscosity is in case of need in own home, monitor oneself seminal fluid situation at any time, assist to evaluate oneself fertility and reproductive system health status, instruct prenatal and postnatal care and sterile diagnoses and treatment, improve the examination rate of infertility, for finding that treatment ahead of time provides possibility ahead of time, provide safeguard for family and social harmony.
Summary of the invention
The invention discloses a kind of relating to analyzes the whether normal fast and convenient decision method of semen viscosity.Adopt the microscope slide of certain rough surface, semen sample is made to start to lean on trickling naturally action of gravity from the fixed position concentration of the microscope slide at certain angle of inclination, check whether seminal fluid trickling front end exceedes markings at the appointed time, judge whether the viscosity of semen sample reaches the minimum standards of male fertility with this result.The qualitative judgement result of this method contributes to analyzing the reason of male sterility.
The applicant of the present invention uses the microscope slide of different roughness, adopt on surface color to mark and significantly drip sampling point and reference line, the seminal fluid primary sample (40～50 μ l) of a droplet post liquefaction is dripped on point sample circle, then microscope slide is tilted on the shelf of certain angle, make semen sample start to lean on trickling naturally action of gravity from dripping sampling point, check whether seminal fluid trickling front end exceedes reference line at the appointed time.If seminal fluid trickling front end exceedes reference line in the stipulated time, then judge that time sample viscosity is normal, namely meet the primary condition of fertility；Whereas if seminal fluid trickles, front end is in the stipulated time not less than reference line, then judge this sample viscosity abnormal (exception), it is possible to be unsatisfactory for the primary condition of fertility, it is proposed that check diagnosis further.This method temporary designations is microscope slide trickling method.
Same sample adopts the WHO any means recommended and microscope slide trickling method to check that whether normal viscosity is simultaneously, after two kinds of methods complete a number of sample, statistical analysis checks result, it is determined that whether the concordance that whether normal the viscosity of two kinds of method acquisitions is has significant difference.
Result shows, the result of determination that whether normal the human semen viscosity of the method acquisition of microscope slide trickling is does not have statistics difference with the WHO dropper method result of determination recommended, there is no statistics difference with the WHO Glass rod method result of determination recommended yet, illustrate that above-mentioned microscope slide trickling method may be used for the quick inspection that whether normal semen sample viscosity is, and assist to evaluate mankind spermatozoon vigor.The more important thing is, use special microscope slide and the support at certain angle of inclination, coordinate the description being easily understood, it is possible to realize experimenter's case limit Self-operating, complete human semen viscosity evaluation, the further examination for infertility provides reference.
The method of the present invention includes herein below:
First the preparation of sample and dyestuff is carried out
1. obtain semen sample according to the suggestion of World Health Organization (WHO)
2. microscope slide: use the medical inspection microscope slide meeting national standard, is processed by certain method, it is thus achieved that rough surface.Draw sample position in microscope slide one end, vertical direction certain distance draws markings
3. tilt stand a: shelf that microscope slide can be made to keep certain inclination or dependence thing, it is ensured that angle of inclination is within the scope of 30～90 degree
4. timer: the timer of second, wrist-watch or intervalometer can be accurate to.
The method that WHO recommends judges that whether semen sample viscosity is normal
Adopting the semen sample that liquefaction is good, mixing is fully.According to dropper method or the Glass rod fiber elongation method of World Health Organization (WHO) 2011 " human seminal fluid checks and treatment of laboratory handbook " load record, judge that whether normal the viscosity of semen sample is by observing the length of wire drawing.When length of string is less than 2 centimetres, namely think that semen sample viscosity is normal, when length of string is more than 2 centimetres, namely think that semen sample viscosity is extremely (abnormal).
Whether microscope slide trickling method semen sample viscosity is normal
For same sample, a droplet semen sample is dropped in rough surface and delimits sample position in advance, it is placed on tilt stand by microscope slide to keep certain gradient, start timing, stipulated time (such as 1min) checks whether the liquid front that semen sample trickles downwards exceedes the markings delimited in advance on microscope slide afterwards, if it does, namely think that semen sample viscosity is normal, without exceeding, namely think that semen sample viscosity is extremely (abnormal).
Statistical analysis
For a number of same batch sample result of determination, two kinds of colouring methods are carried out X 2 test (kappa inspection), and whether the concordance whether normal analyzing viscosity that two kinds of methods obtain has significant difference.P<0.05 illustrates that whether normal two kinds of methods are consistent in analyzing semen sample viscosity, and P>=0.05 item illustrates that whether normal two kinds of methods are inconsistent in analyzing semen sample viscosity.Because dropper method or Glass rod fiber elongation method are the standard methods of world health organisation recommendations, therefore P < proves the method that microscope slide trickling method can be whether normal as judging semen sample viscosity when 0.05.
Conclusion
The semen sample liquefied can at certain angle of inclination, the surface of certain roughness trickle downwards according to gravity direction, viscosity is more low, it is stipulated that time trickling distance is more remote, and viscosity is more high, trickle more slow, in stipulated time trickling apart from more near.
Microscope slide trickling method judges that the result that whether normal semen sample viscosity is does not have statistics difference with dropper method result of determination, there is no statistics difference with Glass rod method result of determination, it is possible to use microscope slide trickling method is for whether normal quickly the judging of semen sample viscosity yet.And, microscope slide trickling method is compared in dropper method and Glass rod method the length of string related to and measures, easy to be accurate, quickness and high efficiency.
Accompanying drawing explanation
Fig. 1: microscope slide essence schematic diagram, circle is sample position, and straight line is markings.
Fig. 2: tilt stand schematic diagram, A face is level, and B face is inclined plane, supports good, and C face is horizontal plane, and the angle in B face and C face is fixing maybe can be regulated.
Detailed description of the invention
Embodiment one microscope slide trickling method for semen sample viscosity whether normal-compare with dropper method
Materials and methods:
1. obtain human seminal fluid's sample
The semen sample adopted provides for volunteer, it is therefore desirable to healthy male volunteers is some, provides semen sample according to following requirement:
1) sample is the sample of normal masturbation ejaculation, it is impossible to for obtaining in condom
2) acquisition of sample, it is necessary to be strict controlled in and ejaculated last time in 3～7 days
3) semen sample is directly collected and is entered in the special specimen cup provided, and fills out ejaculation time and volunteer's numbering, after covering tightly specimen cup lid, delivers to laboratory or appointed place within the time of regulation
4) after once ejaculating, all samples must all be collected in specimen cup, must not have omission, any non-seminal fluid ingredients substance also must not be had to be mixed into
5) leave after health should intact preservation in specimen cup for semen sample, it is prevented that pollute.Note: human seminal fluid belongs to potential source biomolecule dangerous goods, please dispose according to international organization and country's relevant laws and regulations.Advising that insulation next to the skin is carried, temperature is too low or too high possibility affects semen sample viscosity result of determination
6) guarantee that semen sample has liquefied well: liquefaction under semen sample room temperature, can be completed within half an hour, it is therefore proposed that seminal fluid starts test after leaving health half an hour after.Meanwhile, semen sample is not suitable for room temperature and preserves for a long time, it is therefore proposed that seminal fluid completes test in the longest 24 hours after leaving health.
2. microscope slide: the microscope slide (JB/T8230.3-1997) that Henan Shunda Su Ye company limited provides, the rough surface that Ra value is about 12.5 μm is obtained by chemical method for etching, in the color water stroke of microscope slide one end, diameter is the circle of 0.6cm, as sample position, distance vertical direction 2cm place, center of circle fine rule in color water stroke.(such as accompanying drawing 1)
3. tilt stand: middle hollow out or transparent metal or plastic stent, shelf or rely on thing, it is possible to bearing microscope slide and lean to, bottom can spread absorbent paper in advance, in order to absorb the semen sample guttered down.The angle tilted be 65 ° (such as accompanying drawing 2, spread absorbent paper in A face, and the angle in B face and C face is fixed or can regulate)
4. timer: the timer of second, wrist-watch or intervalometer can be accurate to.
5. pipette: be about the wide-mouth footpath disposable plastic pipette of 1.5mm clinically in order to the diameter of the fluid samples such as draw blood, the other end expands, as inhaling ball
6. ruler: be accurate to the transparent ruler of millimeter
Semen sample liquefies
Semen sample leaves room temperature or nearly body temperature after health and preserves, and checks after 30min, confirms liquefied good (judgement that semen sample whether liquefy, with reference to the method that World Health Organization (WHO) 2011 " human seminal fluid checks and treatment of laboratory handbook " load is recorded).The good sample that liquefies completed following test in 23 hours.
Pipette method judges that whether normal semen sample viscosity is
Open for disposable plastic pipette one end is immersed in the semen sample that liquefaction is good, extrudes the enlargement of the other end gently, some semen sample are sucked in plastics pipette, looses one's grip subsequently.Above absorbent paper or waste liquid cylinder, again extrude the enlargement of the pipette other end gently, allow a semen sample drip from pipette opening port, observe drop wire drawing situation, by vertically vertical nigh ruler, measure length of string.If length of string is more than 2cm, be considered as semen sample viscosity abnormal (exception), if length of string not less than 2cm (less than or equal to), be considered as semen sample viscosity normal.When cannot accurately measure too soon such as wire drawing formation, repeatable dropping liquid, repeated measure, until can accurately judge that whether length of string is more than 2cm.
Microscope slide trickling method judges that whether normal semen sample viscosity is
Open for disposable plastic pipette one end is immersed in the semen sample that liquefaction is good, extrudes the enlargement of the other end gently, some semen sample are sucked in plastics pipette, looses one's grip subsequently.Above microscope slide, again extrude the enlargement of the pipette other end gently, allow a semen sample drip in the sample circle drawn in advance microscope slide from pipette opening port.Microscope slide is handled with care on tilt stand, sample spot upper, markings under.Timing at once, whether the liquid front that after 1min, observation semen sample trickles downwards exceedes the markings delimited in advance on microscope slide, if it does, namely think that semen sample viscosity is normal, without exceeding, namely think that semen sample viscosity is extremely (abnormal).The enamel pigment of water-color paintbrush of the circle of labelling sample position is mixed into semen sample, does not affect the judgement that whether normal sample viscosity is, adds color contrast on the contrary, is conducive to whether crossing the judgement of markings.
Statistical analysis
Two kinds of decision methods are carried out statistical analysis for the result of determination of same group of semen sample, adopt SAS statistical analysis software (8.01 version), the kappa inspection of row chi-square criterion, i.e. consistency analysis, P value is calculated according to χ 2 value, P>=0.05 is just it is believed that two kinds of methods are judging that in whether normal human semen viscosity is, there were significant differences, P<0.05 is just it is believed that two kinds of methods are judging do not have difference in human semen viscosity is whether, it is possible to adopt microscope slide trickling method to judge that whether normal semen sample viscosity is.
Result
1. two kinds of methods of table judge that the result that whether normal sample viscosity is collects
Analyze result: Kappa=0.708, ASE=0.137, U=5.188, P=0.0001, illustrate that microscope slide trickling method result of determination is identical with the concordance of plastic suction pipe method result of determination for these 52 samples.
Conclusion
For with a collection of semen sample, microscope slide trickling method is adopted to judge that whether normal its viscosity is one by one, the plastic suction pipe method that WHO recommends simultaneously judges that whether normal viscosity is, two kinds of result of determination are carried out consistency check, result shows that two kinds of methods are identical in the result concordance that whether normal same group of semen sample viscosity of judgement be, it is possible to adopt microscope slide trickling method to judge that whether semen sample viscosity is normal.
Embodiment two microscope slide trickling method for semen sample viscosity whether normal-compare with dropper method
Materials and methods:
1. obtain human seminal fluid's sample
The semen sample adopted provides for volunteer, it is therefore desirable to healthy male volunteers is some, provides semen sample according to following requirement:
1) sample is the sample of normal masturbation ejaculation, it is impossible to for obtaining in condom
2) acquisition of sample, it is necessary to be strict controlled in and ejaculated last time in 3～7 days
3) semen sample is directly collected and is entered in the special specimen cup provided, and fills out ejaculation time and volunteer's numbering, after covering tightly specimen cup lid, delivers to laboratory or appointed place within the time of regulation
4) after once ejaculating, all samples must all be collected in specimen cup, must not have omission, any non-seminal fluid ingredients substance also must not be had to be mixed into
5) leave after health should intact preservation in specimen cup for semen sample, it is prevented that pollute.Note: human seminal fluid belongs to potential source biomolecule dangerous goods, please dispose according to international organization and country's relevant laws and regulations.Advising that insulation next to the skin is carried, temperature is too low or too high possibility affects semen sample viscosity result of determination
6) guarantee that semen sample has liquefied well: liquefaction under semen sample room temperature, can be completed within half an hour, it is therefore proposed that seminal fluid starts test after leaving health half an hour after.Meanwhile, semen sample is not suitable for room temperature and preserves for a long time, it is therefore proposed that seminal fluid completes test in the longest 24 hours after leaving health.
2. microscope slide: the microscope slide (JB/T8230.3-1997) that Henan Shunda Su Ye company limited provides, the rough surface that Ra value is about 25 μm is obtained by corundum polishing method, in the color water stroke of microscope slide one end, diameter is the circle of 0.6cm, as sample position, distance vertical direction 2cm place, center of circle fine rule in color water stroke.
3. tilt stand: middle hollow out or transparent metal or plastic stent, shelf or rely on thing, it is possible to bearing microscope slide and lean to, bottom can spread absorbent paper in advance, and in order to absorb the semen sample guttered down, the angle of inclination is 70 °.
4. timer: the timer of second, wrist-watch or intervalometer can be accurate to.
5. Glass rod: use for laboratory is with the clear glass rod of stirring and evenly mixing, diameter 3mm, and one end processes through high temperature melt and cooling and polishes, and the other end is regardless of, high-temperature sterilization.
6. ruler: be accurate to the transparent ruler of millimeter
Semen sample liquefies
Semen sample leaves room temperature or nearly body temperature after health and preserves, and checks after 30min, confirms liquefied good (judgement that semen sample whether liquefy, with reference to the method that World Health Organization (WHO) 2011 " human seminal fluid checks and treatment of laboratory handbook " load is recorded).The good sample that liquefies completed following test in 23 hours.
Glass rod method judges that whether normal semen sample viscosity is
Smooth for Glass rod one end is immersed in the semen sample that liquefaction is good, provokes semen sample gently, wire drawing situation under sight glass rod, by vertically vertical nigh ruler, measure length of string.If length of string is more than 2cm, be considered as semen sample viscosity abnormal (exception), if length of string not less than 2cm (less than or equal to), be considered as semen sample viscosity normal.As wire drawing formed cannot accurately measure too soon time, can picking, repeated measure, until can accurately judge that whether length of string is more than 2cm.
Microscope slide trickling method judges that whether normal semen sample viscosity is
Open for disposable plastic pipette one end is immersed in the semen sample that liquefaction is good, extrudes the enlargement of the other end gently, some semen sample are sucked in plastics pipette, looses one's grip subsequently.Above microscope slide, again extrude the enlargement of the pipette other end gently, allow a semen sample drip in the sample circle drawn in advance microscope slide from pipette opening port.Microscope slide is handled with care on tilt stand, sample spot upper, markings under.Timing at once, whether the liquid front that after 1min, observation semen sample trickles downwards exceedes the markings delimited in advance on microscope slide, if it does, namely think that semen sample viscosity is normal, without exceeding, namely think that semen sample viscosity is extremely (abnormal).The enamel pigment of water-color paintbrush of the circle of labelling sample position is mixed into semen sample, does not affect the judgement that whether normal sample viscosity is, adds color contrast on the contrary, is conducive to whether crossing the judgement of markings.
Statistical analysis
Two kinds of decision methods are carried out statistical analysis for the result of determination of same group of semen sample, adopt SAS statistical analysis software (8.01 version), the kappa inspection of row chi-square criterion, i.e. consistency analysis, P value is calculated according to χ 2 value, P>=0.05 is just it is believed that two kinds of methods are judging that in whether normal human semen viscosity is, there were significant differences, P<0.05 is just it is believed that two kinds of methods are judging do not have difference in human semen viscosity is whether, it is possible to adopt microscope slide trickling method to judge that whether normal semen sample viscosity is.
Result
2. two kinds of methods of table judge that the result that whether normal sample viscosity is collects
Analyze result: Kappa=0.516, ASE=0.172, U=3.008, P=0.00263, illustrate that microscope slide trickling method result of determination is identical with the concordance of Glass rod method result of determination for above-mentioned 60 samples.
Conclusion
For with a collection of semen sample, microscope slide trickling method is adopted to judge that whether normal its viscosity is one by one, the Glass rod method that WHO recommends simultaneously judges that whether normal viscosity is, two kinds of result of determination are carried out consistency check, result shows that two kinds of methods are identical in the result concordance that whether normal same group of semen sample viscosity of judgement be, it is possible to adopt microscope slide trickling method to judge that whether semen sample viscosity is normal.
Claims (10)
1. from regulation point of sample, semen sample on certain gradient, roughness Ra value scope rough surface in 1.6 to 50 is relied on whether gravity, at the appointed time trickling exceed the markings below certain distance, the application whether normal as judging semen sample viscosity.
2. application according to claim 1, the angular range lower limit of inclination is 30, and the upper limit is 90.
3. application according to claim 1, the point sample weight range lower limit of semen sample is 20l, and range limit is 1ml.
4. application according to claim 1, the time range lower limit that semen sample starts trickling from point of sample is 10 seconds, and range limit is 5min.
5. the distance range lower limit of application according to claim 1, semen sample point of sample and markings is 1cm, and range limit is 20cm.
6. for judging the device that whether normal semen sample viscosity is, wherein said device includes having certain gradient, roughness Ra value scope rough surface in 1.6 to 50, and described rough surface includes point of sample and the markings below described point of sample certain distance.
7. device according to claim 6, described device also includes support, and optionally described support and described rough surface are integral type or split type.
8., for judging the test kit that whether normal semen sample viscosity is, it includes
Device according to any one of claim 6-7；With
Description, from regulation point of sample, the instruction of described description relies on whether gravity, at the appointed time trickling exceed the markings below certain distance according to semen sample, judge that whether normal semen sample viscosity is.
9. test kit according to claim 8, wherein said test kit also includes specimen cup.
10. test kit according to any one of-9 according to Claim 8, wherein said test kit also includes suction pipe.
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