CN109085041A - The method of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility - Google Patents
The method of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility Download PDFInfo
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- CN109085041A CN109085041A CN201810934668.1A CN201810934668A CN109085041A CN 109085041 A CN109085041 A CN 109085041A CN 201810934668 A CN201810934668 A CN 201810934668A CN 109085041 A CN109085041 A CN 109085041A
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 title claims abstract description 14
- 239000012634 fragment Substances 0.000 title claims abstract description 14
- 229910052760 oxygen Inorganic materials 0.000 title claims abstract description 14
- 239000001301 oxygen Substances 0.000 title claims abstract description 14
- 238000000034 method Methods 0.000 title claims abstract description 10
- 238000011156 evaluation Methods 0.000 title claims abstract description 8
- 206010021929 Infertility male Diseases 0.000 title claims abstract description 7
- 208000007466 Male Infertility Diseases 0.000 title claims abstract description 7
- 210000000582 semen Anatomy 0.000 claims abstract description 15
- 208000000509 infertility Diseases 0.000 claims abstract description 10
- 230000036512 infertility Effects 0.000 claims abstract description 10
- 231100000535 infertility Toxicity 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 238000005259 measurement Methods 0.000 abstract description 3
- 230000035558 fertility Effects 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000000694 effects Effects 0.000 description 4
- 208000021267 infertility disease Diseases 0.000 description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 238000010219 correlation analysis Methods 0.000 description 2
- 238000007670 refining Methods 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- ISTBVJOFZNQWJF-UHFFFAOYSA-N 2-(hydroxyamino)-3,7-dihydropurin-6-one Chemical compound O=C1NC(NO)=NC2=C1NC=N2 ISTBVJOFZNQWJF-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 208000032170 Congenital Abnormalities Diseases 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- 230000007698 birth defect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/36—Gynecology or obstetrics
- G01N2800/367—Infertility, e.g. sperm disorder, ovulatory dysfunction
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to DNA fragment with spermatozoon rates, the method of active oxygen and 8-OhdG association evaluation male sterility, measurement is from DNA fragment with spermatozoon rate (DFI) in semen sample, the level of active oxygen (ROS) and 8-OhdG (8-OHdG), by DNA fragment with spermatozoon rate, the level of active oxygen and 8-OhdG is associated with sperm function quality, with sperm DFI=19.26%, ROS=1.91ng/ml and 8-OHdG=50.76ng/l is to judge critical value, as sperm DFI, when ROS and 8-OHdG level is higher than the critical value, judge sperm infertility.
Description
Technical field
The present invention relates to DNA fragment with spermatozoon rates, the method for active oxygen and 8-OhdG association evaluation male sterility.
Background technique
Male sterility annoyings the couple at child-bearing age of the whole world about 10%-15%.The index of existing assessment semen quality relies on
In traditional semen routine analysis, semen routine analysis only detects the indexs such as sperm quantity, viability and form, these indexs
Only reflect basic semen quality, cannot reflect sperm function characteristic, often some patient's Semen routiones are normal, wife's side factor
Normally, it cannot exactly be pregnant.In order to preferably measure male fecundity and prediction reproduction final result, clinic needs accurately to assess
Sperm function.Influence of the integrality of sperm DNA to sperm function is constantly subjected to widely pay close attention to, DNA fragment with spermatozoon rate (DNA
Fragmentation index, DFI), active oxygen (Reactive oxgen species, ROS) and 8-OhdG
(8-Hydroxydeoxyguanosine, 8-OHdG) reflects the degree of injury of hereditary material DNA, there is data expression, essence
The influence of sub- DNA damage can reside in the different phase of fertility, may with rate of fertilization, embryonic development, Implantation Rate, pregnancy rate,
Abortion ratio and the birth defect of offspring etc. are related.
Summary of the invention
The purpose of the present invention is to provide DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation males
The method of infertility.
The purpose of the present invention is achieved through the following technical solutions: DNA fragment with spermatozoon rate, active oxygen and 8- hydroxyl deoxidation bird
The method of glycosides association evaluation male sterility, measurement is from DNA fragment with spermatozoon rate (DFI), active oxygen (ROS) and 8- in semen sample
The level of hydroxy-Guanine (8-OHdG), by the level and sperm of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG
Service quality is associated, critical to judge with sperm DFI=19.26%, ROS=1.91ng/ml and 8-OHdG=50.76ng/l
Value judges sperm infertility when sperm DFI, ROS and 8-OHdG level is higher than the critical value.
The present invention is better able to standard using sperm DFI, ROS and 8-OHdG level as index association evaluation sperm function situation
Really reflect the functional level of sperm.
Detailed description of the invention
Fig. 1 is sperm DFI at sterile group and has an expression in fertility group.
The correlation of Fig. 2 sperm DFI and each parameter of sperm.
The correlation of Fig. 3 refining ROS and each parameter of sperm.
The correlation of Fig. 4 refining 8-OHdG and each parameter of sperm.
The predicted value of Fig. 5 sperm DFI, ROS and 8-OHdG in sterility.
Specific embodiment
1. after taking frozen semen sample to recover, according to " World Health Organization human seminal fluid checks and treatment of laboratory handbook "
The requirement detection sperm concentration and vigor of (the 5th edition).Frozen semen sample 90, wherein 60 have fecundity person's for 60
Semen sample is fertility group；30 semen samples for infertility person, for sterile group.
2. sperm DFI, ROS and 8-OHdG are detected
Sterile group is detected using Sperm Chromatin structural test (SCSA method) and has fertility group sperm DFI.Wherein A liquid master
Want ingredient are as follows: 0.01mol/L Tris-HCl, 0.15mol/L NaCl, 1mmol/L EDTA；B liquid be 0.08mol/L Hcl,
0.5mol/L NaCl, 0.1%Trition X-100；C liquid be 37mmol/L citric acid, 0.126mol/L Na2HPO4,
1mmol/LEDTA,0.15mol/L NaCl,1mg/ml AO.10000 sperms are measured using every part of sperm suspension of fluidic cell,
The ratio of red fluorescence sperm is calculated, is as a result calculated using sperm DNA integrity and reporting system is calculated.Using ELISA
Method detects sterile group and has ROS and 8-OHdG in fertility group sperm horizontal.
3. statistical analysis
Data are for statistical analysis using 22.0 software of SPSS.Measurement data indicates that two groups are compared and examined with t with x ± s,
Correlation uses Spearman rank correlation analysis.P≤0.05 is with statistical significance.
4. experimental result
Sterility group Sperm progressive motility percentage be 11.79 ± 0.83%, total activity power be 22.84 ± 1.24%, essence
Sub- concentration is 73.72 ± 15.64 × 106/ ml, semen volume be 2.87 ± 0.23ml, eupyrene sperm form 6.93 ± 0.51%,
It is 3.35 ± 0.17ng/ml, 8-OHdG is 78.62 ± 4.52ng/l that DFI, which is 32.52 ± 2.02%, ROS,；Before having fertility group
To movement percentage be 41.78 ± 1.15%, total activity power be 64.04 ± 1.49%, sperm concentration be 122.94 ± 6.70 ×
106/ml, semen volume are 3.84 ± 0.15ml, eupyrene sperm form 14.77 ± 0.47%, sperm DFI are 11.05 ± 0.67%,
ROS is that 1.43 ± 0.11ng/ml, 8-OHdG are 34.59 ± 1.81ng/l；Sterility group and there is sperm preceding paragraph between fertility group
Movement percentage, total activity power, sperm concentration, eupyrene sperm form, DFI, ROS and 8-OHdG all have significant difference, lead to
It crosses sensibility and specificity calculates, sterility group and the youden index for having sperm DFI, ROS and 8-OHdG between fertility group
(Yuden Index) is respectively 19.26%, 1.91ng/ml and 50.76ng/l, can be used as determine whether facing for fertility accordingly
Dividing value.It is found by correlation analysis, as in Figure 2-4, sperm DFI and ROS and 8-OHdG positive correlation each other；Sperm
DFI, ROS and 8-OHdG are in respectively significant negative correlativing relation with propulsion percentage, total activity, eupyrene sperm form.As a result,
Sperm DFI and the level of ROS and 8-OHdG is associated with sperm function quality, as sperm DFI > 19.26%, ROS >
When 1.91ng/ml and 8-OHdG > 50.76ng/l, sperm infertility is judged.
Claims (1)
1. the method for DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility, characterized in that survey
Surely DNA fragment with spermatozoon rate (DFI), the level of active oxygen (ROS) and 8-OhdG (8-OHdG) in semen sample are come from,
The level of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG is associated with sperm function quality, with sperm DFI=
19.26%, ROS=1.91ng/ml and 8-OHdG=50.76ng/l is to judge critical value, when sperm DFI, ROS and 8-OHdG water
When putting down higher than the critical value, sperm infertility is judged.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810934668.1A CN109085041A (en) | 2018-08-16 | 2018-08-16 | The method of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810934668.1A CN109085041A (en) | 2018-08-16 | 2018-08-16 | The method of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility |
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| Publication Number | Publication Date |
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| CN109085041A true CN109085041A (en) | 2018-12-25 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201810934668.1A Pending CN109085041A (en) | 2018-08-16 | 2018-08-16 | The method of DNA fragment with spermatozoon rate, active oxygen and 8-OhdG association evaluation male sterility |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110967230A (en) * | 2019-11-22 | 2020-04-07 | 珠海高瑞特医疗科技有限公司 | Method and kit for measuring content of active oxygen in sperm |
| CN113584148A (en) * | 2021-06-30 | 2021-11-02 | 吉林大学 | Specific marker screening method for azoospermia and severe oligospermia detection |
-
2018
- 2018-08-16 CN CN201810934668.1A patent/CN109085041A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110967230A (en) * | 2019-11-22 | 2020-04-07 | 珠海高瑞特医疗科技有限公司 | Method and kit for measuring content of active oxygen in sperm |
| CN113584148A (en) * | 2021-06-30 | 2021-11-02 | 吉林大学 | Specific marker screening method for azoospermia and severe oligospermia detection |
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| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181225 |
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