CN103571914B - Obtain acquisition method and the device of nucleic acid clinical samples - Google Patents
Obtain acquisition method and the device of nucleic acid clinical samples Download PDFInfo
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- CN103571914B CN103571914B CN201310068049.6A CN201310068049A CN103571914B CN 103571914 B CN103571914 B CN 103571914B CN 201310068049 A CN201310068049 A CN 201310068049A CN 103571914 B CN103571914 B CN 103571914B
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- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 19
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 19
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 19
- 238000000034 method Methods 0.000 title claims abstract description 11
- 239000012528 membrane Substances 0.000 claims abstract description 24
- 239000008367 deionised water Substances 0.000 claims abstract description 6
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000007984 Tris EDTA buffer Substances 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 3
- 239000013078 crystal Substances 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- 239000003365 glass fiber Substances 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 229910052710 silicon Inorganic materials 0.000 claims description 3
- 239000010703 silicon Substances 0.000 claims description 3
- 238000002604 ultrasonography Methods 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 abstract 2
- 239000012530 fluid Substances 0.000 abstract 2
- 238000001914 filtration Methods 0.000 abstract 1
- 239000000443 aerosol Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000010339 medical test Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
技术领域technical field
本发明涉及一种获取核酸临床标本的采集方法及装置。The invention relates to a collection method and device for obtaining nucleic acid clinical specimens.
背景技术Background technique
医学检测时，经常需要采集患者的核酸临床标本。现有的采集方法多为采集患者外周血3-5毫升或者组织标本1-2mg，通过试剂盒提取核酸，获得核酸临床标本。但是还没有很好的方法从患者呼出气体中收集和提取游离的核酸标本。During medical testing, it is often necessary to collect nucleic acid clinical specimens from patients. Most of the existing collection methods are to collect 3-5 ml of peripheral blood or 1-2 mg of tissue samples from patients, and extract nucleic acids through kits to obtain nucleic acid clinical samples. However, there is no good method to collect and extract free nucleic acid samples from patients' exhaled breath.
发明内容Contents of the invention
本发明的发明目的在于提供一种获取核酸临床标本的采集方法，不需采血、操作简便，采集效果好。The object of the present invention is to provide a collection method for obtaining nucleic acid clinical specimens, which does not require blood collection, is easy to operate, and has good collection effects.
基于同一发明构思，本发明具有两个独立的技术方案：Based on the same inventive concept, the present invention has two independent technical solutions:
1.一种获取核酸临床标本的采集方法，其特征在于：通过采集装置采集患者呼出的空气，通过采集装置的滤膜对患者呼出的空气过滤，将滤膜浸泡于TE缓冲液或去离子水中，利用超声把滤膜上的收集物溶出，收集溶出液，溶出液为核酸临床标本。1. A collection method for obtaining nucleic acid clinical specimens, characterized in that: the air exhaled by the patient is collected by the collection device, the air exhaled by the patient is filtered by the filter membrane of the collection device, and the filter membrane is soaked in TE buffer solution or deionized water , using ultrasound to dissolve the collected matter on the filter membrane, and collect the eluate, which is a clinical specimen of nucleic acid.
将滤膜浸泡于TE缓冲液或去离子水中时，需1000g离心10分钟。When soaking the filter membrane in TE buffer or deionized water, centrifuge at 1000g for 10 minutes.
收集溶出液时，需13000g离心10分钟。When collecting the eluate, it needs to be centrifuged at 13000g for 10 minutes.
通过采集装置采集患者呼出的空气5-10L。Collect 5-10L of air exhaled by the patient through the collection device.
2.用于实现上述方法的采集装置，其特征在于：空气收集管、过滤管和溶出液收集管由上至下依次插接，过滤管内设有滤膜，过滤管的底端设有开口，开口处设有封闭盖，过滤管的顶端设有管盖。2. The acquisition device for realizing the above method is characterized in that: the air collection tube, the filter tube and the eluate collection tube are plugged sequentially from top to bottom, the filter tube is provided with a filter membrane, and the bottom end of the filter tube is provided with an opening, A closing cover is provided at the opening, and a tube cover is provided at the top of the filter tube.
空气收集管的上部、过滤管的下部均呈锥形。Both the upper part of the air collecting pipe and the lower part of the filter pipe are tapered.
过滤管底端的开口与封闭盖螺纹连接。The opening at the bottom end of the filter tube is threadedly connected with the closure cap.
本发明具有的有益效果：The beneficial effect that the present invention has:
微生物或其释放的核酸类物质在空气中形成气溶胶，当气溶胶通过滤膜时，其中的大分子核酸被截留在滤膜上。用去离子水或TE缓冲液浸润滤膜，通过超声将滤膜上的收集物溶出，收集溶出液即获得核酸临床标本。本发明不需采血，只需患者呼气即可实现核酸临床标本的采集，操作简单、快速、高效、无创伤，本发明采集装置不仅可应用于结核菌检测等医学检测，还可以广泛应用于环境监测。Microorganisms or the nucleic acid substances they release form aerosols in the air, and when the aerosol passes through the filter membrane, the macromolecular nucleic acids in it are trapped on the filter membrane. Soak the filter membrane with deionized water or TE buffer solution, dissolve the collection on the filter membrane by ultrasound, and collect the eluate to obtain the nucleic acid clinical specimen. The present invention does not require blood collection, and only needs to be exhaled by the patient to realize the collection of nucleic acid clinical specimens. The operation is simple, fast, efficient, and non-invasive. The collection device of the present invention can not only be applied to medical detection such as tuberculosis detection, but also can be widely used in environmental monitoring.
附图说明Description of drawings
图1为本发明采集装置的结构示意图；Fig. 1 is the structural representation of collecting device of the present invention;
图2为本发明采集装置过滤管的结构示意图。Fig. 2 is a schematic structural view of the filter tube of the collection device of the present invention.
具体实施方式Detailed ways
如图1所示，采集装置包括空气收集管1、过滤管2和溶出液收集管3，空气收集管1、过滤管2和溶出液收集管3由上至下依次插接，过滤管的底端与溶出液收集管的底部之间留有空间。空气收集管1用于接收患者呼出的气体，也可与其他供气设备连接。空气收集管1的上部、过滤管2的下部均呈锥形。如图2所示，过滤管内设有滤膜5，滤膜由玻璃纤维、硅晶纤维、TCLP或硅胶制成。滤膜5置于托架6上，托架6为网状结构，对滤膜5起到支撑作用，防止滤膜5脱落。过滤管的顶端设有管盖4，过滤管的底端设有开口，开口处设有封闭盖7，过滤管底端的开口外侧设有外螺纹，封闭盖内侧设有内螺纹，过滤管底端开口与封闭盖螺纹连接，过滤管底端的开口还可与负压引器等设备连接。As shown in Figure 1, the collection device includes an air collection tube 1, a filter tube 2, and an eluate collection tube 3, and the air collection tube 1, filter tube 2, and eluate collection tube 3 are inserted in sequence from top to bottom, and the bottom of the filter tube is Leave a space between the end and the bottom of the eluate collection tube. The air collection tube 1 is used to receive the gas exhaled by the patient, and can also be connected with other gas supply equipment. The upper part of the air collection pipe 1 and the lower part of the filter pipe 2 are both tapered. As shown in Figure 2, a filter membrane 5 is arranged inside the filter tube, and the filter membrane is made of glass fiber, silicon crystal fiber, TCLP or silica gel. The filter membrane 5 is placed on the bracket 6, and the bracket 6 has a mesh structure, which supports the filter membrane 5 and prevents the filter membrane 5 from falling off. The top of filter tube is provided with cap 4, and the bottom of filter tube is provided with opening, and opening is provided with closure cap 7, and the outside of the opening of filter tube bottom is provided with external thread, and the inside of closure cap is provided with internal thread, and the bottom of filter tube The opening is threadedly connected with the closing cap, and the opening at the bottom end of the filter tube can also be connected with equipment such as a negative pressure guide.
采集核酸标本时，将空气收集管1、过滤管2插接，此时不要使用封闭盖7；患者深吸气后，用空气收集管封闭患者口部，患者用力呼气，重复3次，收集5-10L患者呼气；然后旋紧封闭盖7，撤去空气收集管1，将过滤管2插入溶出液收集管内；向过滤管中加入800微升的TE缓冲液或去离子水，盖上管盖，1000g离心10分钟，使液体完全浸润滤膜5；打开管盖4，插入超声探头，经短时间超声把滤膜5上的收集物溶出；旋下封闭盖7，盖上管盖4，将滤管放入一个新的溶出液收集管；13000g高速离心10分钟，溶出液收集管内收集的溶出液即为待测核酸标本，低温保存。When collecting nucleic acid specimens, connect the air collection tube 1 and filter tube 2, and do not use the closing cap 7 at this time; after the patient inhales deeply, close the mouth of the patient with the air collection tube, and the patient exhales forcefully, repeat 3 times, and collect 5-10L patient exhales; then tighten the closure cap 7, remove the air collection tube 1, insert the filter tube 2 into the eluate collection tube; add 800 microliters of TE buffer or deionized water to the filter tube, cover the tube Cover, and centrifuge at 1000g for 10 minutes to make the liquid completely infiltrate the filter membrane 5; open the tube cover 4, insert an ultrasonic probe, and dissolve the collection on the filter membrane 5 by ultrasonication for a short time; unscrew the sealing cover 7, cover the tube cover 4, Put the filter tube into a new eluate collection tube; centrifuge at a high speed of 13000g for 10 minutes, the eluate collected in the eluate collection tube is the nucleic acid sample to be tested, and store it at low temperature.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310068049.6A CN103571914B (en) | 2013-02-25 | 2013-02-25 | Obtain acquisition method and the device of nucleic acid clinical samples |
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| CN201310068049.6A CN103571914B (en) | 2013-02-25 | 2013-02-25 | Obtain acquisition method and the device of nucleic acid clinical samples |
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| CN103571914A CN103571914A (en) | 2014-02-12 |
| CN103571914B true CN103571914B (en) | 2015-11-25 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| GB2548122B (en) * | 2016-03-08 | 2018-09-12 | Univ Of Northumbria At Newcastle | Exhaled breath condensate collection device and a kit of parts therefor |
| CN109207340B (en) * | 2017-06-30 | 2022-08-12 | 开启基因股份有限公司 | Nucleic acid extraction components |
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2013
- 2013-02-25 CN CN201310068049.6A patent/CN103571914B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| 呼吸道鼻病毒、SARS 病毒的Teflo 滤膜采集及RT-PCR 检测的初步研究;韩黎等;《军医进修学院学报》;20031231;第24卷(第4期);249-251 * |
| 采集呼气标本的时间对检测胃幽门螺杆菌感染检出率的影响;侯丽英;《山西医药杂志》;20060930;第35卷(第9期);836-837 * |
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