CN113416772A - 精子dna碎片化质控品的组成配方 - Google Patents
精子dna碎片化质控品的组成配方 Download PDFInfo
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- 238000013467 fragmentation Methods 0.000 title claims abstract description 30
- 238000006062 fragmentation reaction Methods 0.000 title claims abstract description 30
- 238000003908 quality control method Methods 0.000 title claims abstract description 23
- 239000000203 mixture Substances 0.000 title claims abstract description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 26
- 235000013905 glycine and its sodium salt Nutrition 0.000 claims abstract description 16
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000007836 KH2PO4 Substances 0.000 claims abstract description 13
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 13
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 13
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims abstract description 13
- 239000011780 sodium chloride Substances 0.000 claims abstract description 13
- 238000009472 formulation Methods 0.000 claims 3
- 239000003814 drug Substances 0.000 abstract description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 208000007466 Male Infertility Diseases 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
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- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
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Abstract
本发明公开了精子DNA碎片化质控品的组成配方,包括浓度为1E+6~1E+8个/mL的正常细胞、浓度为1E+6~1E+8个/mL的DNA碎片化细胞、浓度为1.2~8.7mmol/L的KCl、浓度为0.5~5.6mmol/L的KH2PO4、浓度为0.05~0.25mol/L的NaCl、浓度为3.4~12.2mmol/L的Na2HPO4‑7H2O、浓度为8.6~32.1mmol/L的C11H16N8O8、浓度为12.3~36.5mmol/L的C2H5NO2、浓度为1~4mmol/L的EDTA、质量体积比浓度为0.3%~2%的BSA,涉及医疗领域。本发明型通过添加C11H16N8O8、C2H5NO2、EDTA三种药剂,再加上正常细胞与碎片化细胞进行混合,能够使得质控品能够更加稳定,使得保存时间更长,同时通过将正常细胞与碎片化细胞进行不同比例混合,可以控制DNA碎片化值,从而比精子样本大大减小批间差异,从而能够大规模进行临床试验。
Description
技术领域
本发明涉及医疗领域,具体为精子DNA碎片化质控品的组成配方。
背景技术
精子细胞DNA碎片化是影响男性不育的最重要因素之一,还可能导致反复流产和助孕(IVF/ICSI)治疗成功率低,精子DNA碎片化程度(DFI)被认为是一个新的评价精子质量和预测生育能力的指标,在众多医院中已经开展了对精子DFI的检测,因此,本发明的用于精子DFI检测的质控品具有非常重要的意义。
但是现如今的精子DNA碎片化检测用质控品主要是采用真实精子样本作为实验室内部质控,目前市面上还没有可用于精子DNA碎片化检测的标准化的质控品,而传统用精子样本作为质控品存在的问题有:无法在实验室间进行质控,保存困难,易受环境影响,重复性差,DNA碎片化值无法控制,批间差异较大,无法大规模在临床上进行推广等问题。
发明内容
本发明的目的在于:为了解决的问题,提供精子DNA碎片化质控品的组成配方。
为实现上述目的,本发明提供如下技术方案:精子DNA碎片化质控品的组成配方,包括浓度为1E+6~1E+8个/mL的正常细胞、浓度为1E+6~1E+8个/mL的DNA碎片化细胞、浓度为1.2~8.7mmol/L的KCl、浓度为0.5~5.6mmol/L的KH2PO4、浓度为0.05~0.25mol/L的NaCl、浓度为3.4~12.2mmol/L的Na2HPO4-7H2O、浓度为8.6~32.1mmol/L的C11H16N8O8、浓度为12.3~36.5mmol/L的C2H5NO2、浓度为1~4mmol/L的EDTA、质量体积比浓度为0.3%~2%的BSA。
优选地,所述浓度为1E+6个/mL的正常细胞、浓度为1E+6个/mL的DNA碎片化细胞、浓度为1.2mmol/L的KCl、浓度为0.5mmol/L的KH2PO4、浓度为0.05mol/L的NaCl、浓度为3.4mmol/L的Na2HPO4-7H2O、浓度为8.6mmol/L的C11H16N8O8、浓度为12.3mmol/L的C2H5NO2、浓度为1mmol/L的EDTA、质量体积比浓度为0.2%的BSA。
优选地,所述浓度为1E+8个/mL的正常细胞、浓度为1E+8个/mL的DNA碎片化细胞、浓度为8.7mmol/L的KCl、浓度为5.6mmol/L的KH2PO4、浓度为0.25mol/L的NaCl、浓度为12.2mmol/L的Na2HPO4-7H2O、浓度为32.1mmol/L的C11H16N8O8、浓度为36.5mmol/L的C2H5NO2、浓度为4mmol/L的EDTA、质量体积比浓度为3%的BSA。
优选地,所述浓度为1E+7个/mL的正常细胞、浓度为1E+7个/mL的DNA碎片化细胞、浓度为4.3mmol/L的KCl、浓度为3.5mmol/L的KH2PO4、浓度为0.125mol/L的NaCl、浓度为8mmol/L的Na2HPO4-7H2O、浓度为20mmol/L的C11H16N8O8、浓度为25mmol/L的C2H5NO2、浓度为2.5mmol/L的EDTA、质量体积比浓度为0.25%的BSA。
与现有技术相比,本发明的有益效果是:本发明通过添加C11H16N8O8、C2H5NO2、DTA三种药剂,再加上正常细胞与碎片化细胞进行混合,能够使得质控品能后更加稳定,使得保存时间更长,同时通过将正常细胞与碎片化细胞进行不同比例混合,可以控制DNA碎片化值,从而比精子样本大大减小批间差异,从而能够大规模进行临床试验。
具体实施方式
下面将结合本发明实施例中的,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
精子DNA碎片化质控品的组成配方,包括浓度为1E+6~1E+8个/mL的正常细胞、浓度为1E+6~1E+8个/mL的DNA碎片化细胞、浓度为1.2~8.7mmol/L的KCl、浓度为0.5~5.6mmol/L的KH2PO4、浓度为0.05~0.25mol/L的NaCl、浓度为3.4~12.2mmol/L的Na2HPO4-7H2O、浓度为8.6~32.1mmol/L的C11H16N8O8、浓度为12.3~36.5mmol/L的C2H5NO2、浓度为1~4mmol/L的EDTA、质量体积比浓度为0.3%~2%的BSA。
本发明通过添加C11H16N8O8、C2H5NO2、EDTA三种药剂,再加上正常细胞与碎片化细胞进行混合,能够使得质控品能后更加稳定。
实施例一
浓度为1E+6个/mL的正常细胞、浓度为1E+6个/mL的DNA碎片化细胞、浓度为1.2mmol/L的KCl、浓度为0.5mmol/L的KH2PO4、浓度为0.05mol/L的NaCl、浓度为3.4mmol/L的Na2HPO4-7H2O、浓度为8.6mmol/L的C11H16N8O8、浓度为12.3mmol/L的C2H5NO2、浓度为1mmol/L的EDTA、质量体积比浓度为0.2%的BSA。
添加的浓度较小的试剂,会导致质控品存放时间较短,不易于保存。
实施例二
浓度为1E+8个/mL的正常细胞、浓度为1E+8个/mL的DNA碎片化细胞、浓度为8.7mmol/L的KCl、浓度为5.6mmol/L的KH2PO4、浓度为0.25mol/L的NaCl、浓度为12.2mmol/L的Na2HPO4-7H2O、浓度为32.1mmol/L的C11H16N8O8、浓度为36.5mmol/L的C2H5NO2、浓度为4mmol/L的EDTA、质量体积比浓度为3%的BSA。
添加浓度较高的试剂并配合浓度较高的正常细胞与碎片化细胞,导致质控品的细胞激素损坏,DNA碎片化值不准确,而且保存时间较短。
实施例三
浓度为1E+7个/mL的正常细胞、浓度为1E+7个/mL的DNA碎片化细胞、浓度为4.3mmol/L的KCl、浓度为3.5mmol/L的KH2PO4、浓度为0.125mol/L的NaCl、浓度为8mmol/L的Na2HPO4-7H2O、浓度为20mmol/L的C11H16N8O8、浓度为25mmol/L的C2H5NO2、浓度为2.5mmol/L的EDTA、质量体积比浓度为0.25%的BSA。
添加合适浓度的试剂与合适浓度正常细胞与碎片化细胞,能够使得质控品更加稳定,保存时间更长,使得DNA碎片化值更加准确。
对于本领域技术人员而言,显然本发明不限于上述示范性实施例的细节,而且在不背离本发明的精神或基本特征的情况下,能够以其他的具体形式实现本发明。因此,无论从哪一点来看,均应将实施例看作是示范性的,而且是非限制性的,本发明的范围由所附权利要求而不是上述说明限定,因此旨在将落在权利要求的等同要件的含义和范围内的所有变化囊括在本发明内。不应将权利要求中的任何标记视为限制所涉及的权利要求。
Claims (4)
1.精子DNA碎片化质控品的组成配方,包括浓度为1E+6~1E+8个/mL的正常细胞、浓度为1E+6~1E+8个/mL的DNA碎片化细胞、浓度为1.2~8.7mmol/L的KCl、浓度为0.5~5.6mmol/L的KH2PO4、浓度为0.05~0.25mol/L的NaCl、浓度为3.4~12.2mmol/L的Na2HPO4-7H2O、浓度为8.6~32.1mmol/L的C11H16N8O8、浓度为12.3~36.5mmol/L的C2H5NO2、浓度为1~4mmol/L的EDTA、质量体积比浓度为0.3%~2%的BSA。
2.根据权利要求1所述的精子DNA碎片化质控品的组成配方,其特征在于:所述浓度为1E+6个/mL的正常细胞、浓度为1E+6个/mL的DNA碎片化细胞、浓度为1.2mmol/L的KCl、浓度为0.5mmol/L的KH2PO4、浓度为0.05mol/L的NaCl、浓度为3.4mmol/L的Na2HPO4-7H2O、浓度为8.6mmol/L的C11H16N8O8、浓度为12.3mmol/L的C2H5NO2、浓度为1mmol/L的EDTA、质量体积比浓度为0.2%的BSA。
3.根据权利要求1所述的精子DNA碎片化质控品的组成配方,其特征在于:所述浓度为1E+8个/mL的正常细胞、浓度为1E+8个/mL的DNA碎片化细胞、浓度为8.7mmol/L的KCl、浓度为5.6mmol/L的KH2PO4、浓度为0.25mol/L的NaCl、浓度为12.2mmol/L的Na2HPO4-7H2O、浓度为32.1mmol/L的C11H16N8O8、浓度为36.5mmol/L的C2H5NO2、浓度为4mmol/L的EDTA、质量体积比浓度为3%的BSA。
4.根据权利要求1所述的精子DNA碎片化质控品的组成配方,其特征在于:所述浓度为1E+7个/mL的正常细胞、浓度为1E+7个/mL的DNA碎片化细胞、浓度为4.3mmol/L的KCl、浓度为3.5mmol/L的KH2PO4、浓度为0.125mol/L的NaCl、浓度为8mmol/L的Na2HPO4-7H2O、浓度为20mmol/L的C11H16N8O8、浓度为25mmol/L的C2H5NO2、浓度为2.5mmol/L的EDTA、质量体积比浓度为0.25%的BSA。
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
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US7070917B1 (en) * | 1999-03-05 | 2006-07-04 | Preben Christensen | Determination of sperm concentration and viability for artificial insemination |
CN103335934A (zh) * | 2013-06-28 | 2013-10-02 | 浙江星博生物科技有限公司 | 一种基于流式细胞术的精子活率检测试剂 |
WO2017156843A1 (zh) * | 2016-03-16 | 2017-09-21 | 四川大学华西第二医院 | 一种精子质量评估的试剂盒及其使用方法 |
CN111307696A (zh) * | 2020-03-19 | 2020-06-19 | 浙江星博生物科技股份有限公司 | 用于检测精子dna碎片率的方法和试剂盒 |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7070917B1 (en) * | 1999-03-05 | 2006-07-04 | Preben Christensen | Determination of sperm concentration and viability for artificial insemination |
CN103335934A (zh) * | 2013-06-28 | 2013-10-02 | 浙江星博生物科技有限公司 | 一种基于流式细胞术的精子活率检测试剂 |
WO2017156843A1 (zh) * | 2016-03-16 | 2017-09-21 | 四川大学华西第二医院 | 一种精子质量评估的试剂盒及其使用方法 |
CN111307696A (zh) * | 2020-03-19 | 2020-06-19 | 浙江星博生物科技股份有限公司 | 用于检测精子dna碎片率的方法和试剂盒 |
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