CN104849122A - Long-acting tumor tissue exudate sample preservation solution - Google Patents
Long-acting tumor tissue exudate sample preservation solution Download PDFInfo
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- CN104849122A CN104849122A CN201510295106.3A CN201510295106A CN104849122A CN 104849122 A CN104849122 A CN 104849122A CN 201510295106 A CN201510295106 A CN 201510295106A CN 104849122 A CN104849122 A CN 104849122A
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Abstract
The invention discloses a long-acting tumor tissue exudate sample preservation solution. The sample preservation solution consists of components with the following proportions: 3.69-3.76g/L ethylene diamine tetraacetic acid, 38-42mg/L isobutanol, 9.5-10.5ml/L formaldehyde and a 0.1-1mol/L phosphate buffer solution. By adopting the prepared long-acting tumor tissue exudate sample preservation solution disclosed by the invention, a stable environment can be provided for a tumor tissue exudate sample within 90 days, the coagulation and precipitation of tangible materials in an exudate can be prevented, and the effects of removing mucus in the sample, reducing the viscosity and preventing corrosion can also be achieved so as to ensure that the absorbance of the sample is in a controllable state, and the accuracy of heme detection of tumor cells can be ensured.
Description
Technical field
The present invention relates to a kind of Sample preservation liquid, specifically a kind of long-acting tumor tissues sepage Sample preservation liquid.
Background technology
Inflammation is the seventh-largest feature of tumour.The activation of oncogene can cause the generation of inflammatory molecule and the gathering of inflammatory cell.One group of Cytokine protein, comprises IL-1, and the inflammatory factors such as IL-6, TNF and RANKL activate the key transcription factor NF-kappaB in downstream and promote inflammation.Liquid in tumor blood vessels and tumour cell composition are entered by vascular wall organizes the process of interstitial, body cavity, mucomembranous surface and body surface to be called that tumprigenicity oozes out.The liquid oozed out and cell are generically and collectively referred to as tumprigenicity exudate or transudate.It is the distinctive change of tumprigenicity inflammation most that tumprigenicity centered by vascular reaction oozes out pathology, containing higher protein and more cell component and their disintegration product in inflammatory exudate.This process medium vessels reaction main manifestations is hemodynamic responses (inflammatory congestion), vasopermeability increases (inflammatory oozes out), liquid oozes out and oozes out (inflammatory infiltration) with cell.Due to inflammatory infiltration around tumor tissues, tumprigenicity oozes out to be organized in sepage and may contain tumor tissue cell's hemn, whether it is in histocyte cancerates process, produce a kind of causality biological marker factor, and organize in sepage significant to organizing the qualification of whether cancerating containing tumor tissue cell's hemn.Detect a kind of new method that tumor tissue cell's hemn is examination malignant tumour.Tumor tissues sepage can detect the existence of tumor tissue cell's hemn as direct biological media.
Gather and detect in tumor tissues sepage in analytic process, there is following difficult point: (1) is due to inflammatory reaction, organize in sepage and destroy the thromboplastin and a large amount of cast-off cells of releasing containing protein, sugar, triglyceride, cholesterol and fibrinogen, these materials may form interference to the detection of tumor tissue cell's hemn.If organize sepage sample muddiness heavier, or contained above-mentioned substance is too much, and the absorbance height causing sample differs, and automated analysis is detected cannot carry out, or testing result misalignment.After tumor tissues sepage sampling (2), send and test before, sample needs to preserve a period of time in conserving liquid.Sometimes, in order to dynamic monitoring and check need, same sample also needs to preserve the longer time.(3) tumor tissue cell's hemn is the object detected, and it is a kind of enzyme sample material, very unstable.For solving above three problems, need a kind ofly to provide stable environment for tumor tissues sepage sample, tangible mass sets and precipitation in anti-impervious liquid, mucus in removing sample is to reduce viscosity and to have preservative efficacy, and make the absorbance of sample be in controllable state, improve the preconditioning technique analyzing precision and the reliability detected.The conserving liquid that development can meet above demand is problem demanding prompt solution.Through retrieval, there is no tumor tissues sepage Sample preservation liquid at present, cause tumor tissues sepage to preserve, tumor tissue cell's hemn detects and cannot carry out.The invention provides a kind of sample long-acting tumor tissues sepage conserving liquid that can meet tumor tissue cell's hemn analysis and detect.
Summary of the invention
The object of the present invention is to provide a kind of long-acting tumor tissues sepage Sample preservation liquid used in tumor tissue cell's hemn testing process.Long-acting tumor tissues sepage Sample preservation liquid prepared by the present invention, can for tumor tissues sepage sample provides stable environment in 90 days, tangible mass sets and precipitation in anti-impervious liquid, and the mucus had in removing sample reduces viscosity and antisepsis, make the absorbance of sample be in controllable state, ensure the accuracy that tumour cell hemn detects.
The present invention is by the following technical solutions:
A kind of long-acting tumor tissues sepage Sample preservation liquid, it is characterized in that, it is made up of 3.69 ~ 3.76g/L disodium ethylene diamine tetraacetate, 38 ~ 42mg/L isobutyl alcohol, 9.5 ~ 10.5ml/L formaldehyde and 0.1mol/L ~ 1mol/L phosphate buffer; The pH value of described phosphate buffer is 6.2 ~ 7.8.
Preferably, described long-acting tumor tissues sepage Sample preservation liquid is that 7.4 phosphate buffers form by the disodium ethylene diamine tetraacetate of 3.72g/L, the isobutyl alcohol of 40mg/L, the formaldehyde of 10ml/L and 0.5mol/L pH value.
The using method of the present invention's long-acting tumor tissues sepage Sample preservation liquid is as follows: select suitable sampling thief, and in detection position wiping gently, what obtain local organizes sepage.Discharge rate is 0.5 ~ 1ml, generally all moistening for degree with sampling swab.After sampling, swab is inserted in conserving liquid 2ml and stir gently, the Sample preservation liquid of sepage and the present invention of organizing that swab obtains is mixed, obtained sample liquid normal temperature is preserved, is sent and test.
Mechanism and the beneficial effect of the present invention's long-acting tumor tissues sepage Sample preservation liquid are as follows: phosphate-buffered liquid system, provides stable acid or alkali environment.Disodium ethylene diamine tetraacetate is a kind of stabilizing agent, increases the stability of the tumor tissue cell's hemn in sample; It also has anti-freezing and prevents the function of tangible species precipitate; Simultaneously it or a kind of complexing agent, the interference of the metallic ion that may exist in minimizing sample.Isobutyl alcohol is a kind of organic cosolvent, also have and eliminate the effect that mucus reduces sample liquid viscosity, contribute to again the organism dissolving and extract in sample, reduce the interference of the materials such as protein, sugar, triglyceride, cholesterol and the fibrinogen that may exist in sample.Formaldehyde has anti-corrosion function.Compatibility of the present invention is reasonable; preparation method is simple and practical; the product of preparation can properly preserve tumor tissues sepage sample 90 days at normal temperatures; tangible mass sets and precipitation in anti-impervious liquid; and the mucus had in removing sample reduces viscosity and antisepsis; effectively protect testing goal thing---the stability of tumor tissue cell's hemn, reject disturbing factor, ensure the precision detected.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail.
Embodiment 1
Tumor tissues sepage Sample preservation formula of liquid: 3.69g/L disodium ethylene diamine tetraacetate, 38mg/L isobutyl alcohol, 9.5ml/L formaldehyde and 0.1mol/L pH value are the phosphate buffer composition of 7.8.
The preparation method of above-mentioned Sample preservation liquid:
(1) the phosphate buffer 1 L that 0.1mol/L pH value is 7.8 is prepared;
(2) disodium ethylene diamine tetraacetate 3.69g is joined in above-mentioned phosphate buffer, heating for dissolving in 60 ± 3 DEG C of water-baths;
(3) isobutyl alcohol 38mg is added in the obtained solution of step (2), fully stir and make it to dissolve completely;
(4) formaldehyde 9.5ml is added in the obtained solution of step (3), stir and make it mixing.
Embodiment 2
Tumor tissues sepage Sample preservation formula of liquid: the formaldehyde of the disodium ethylene diamine tetraacetate of 3.72g/L, the isobutyl alcohol of 40mg/L, 10ml/L and 0.2mol/L pH value are 7.4 phosphate buffers.
The preparation method of above-mentioned Sample preservation liquid:
(1) preparing 0.2mol/L pH value is the phosphate buffer of 7.4;
(2) disodium ethylene diamine tetraacetate 3.72g is joined in above-mentioned phosphate buffer, heating for dissolving in 60 ± 3 DEG C of water-baths;
(3) isobutyl alcohol 40mg is added in the obtained solution of step (2), fully stir and make it to dissolve completely;
(4) formaldehyde 10ml is added in the obtained solution of step (3), stir and make it mixing.
Embodiment 3
Tumor tissues sepage Sample preservation formula of liquid: 3.76g/L disodium ethylene diamine tetraacetate, 42mg/L isobutyl alcohol, 10.5ml/L formaldehyde and 1mol/L pH value are the phosphate buffer composition of 6.2.
The preparation method of above-mentioned Sample preservation liquid:
(1) preparing 1mol/L pH value is the phosphate buffer of 6.2;
(2) disodium ethylene diamine tetraacetate 3.76g is joined in above-mentioned phosphate buffer, heating for dissolving in 60 ± 3 DEG C of water-baths;
(3) isobutyl alcohol 42mg is added in the obtained solution of step (2), fully stir and make it to dissolve completely;
(4) formaldehyde 10.5ml is added in the obtained solution of step (3), stir and make it mixing.
Embodiment 4
Validity is tested
1, test method
(1) testing equipment: full-automatic tumor tissue cell hemn detects analyser.
(2) reagent is detected: organize sepage tumor tissue cell hemn quantitative detecting reagent.
(3) sample liquid is detected: tumor tissues sepage sample 0.5ml adds the interior mixing of tumor tissues sepage Sample preservation liquid 2ml prepared by embodiment 2, and normal temperature saves backup.
(4) detection method: after obtained sample liquid, (0 day) measures sample liquid inner tumour cell hemn content immediately, after this detects 1 time every 15 days to same sample liquid, to the 90th day.
(5) operator scheme: add sample liquid 200 μ L to be checked in color comparison tube, then add and organize sepage tumor tissue cell hemn quantitative detecting reagent 200 μ L, 37 DEG C of incubations make reagent and sample fully react.On the reaction time point of 144 seconds, read the absorbance of nitrite ion after this sample and reagent reacting with photoelectric colorimetry, read the content (μ g/L) of corresponding tumor tissue cell's hemn with this absorbance by the typical curve that instrument sets.
(6) statistical procedures: adopt variance test to carry out otherness to testing result and compare.
2, the making of experimental animal sample liquid:
(1) foundation of bringing out property nasopharyngeal carcinoma small white mouse animal model: get about 120 g big white mouse, after etherization, with No. 8 syringe needles polishing needle point, insert gently from prenaris, needle point can reach nasopharyngeal cavity; DEN 6.7 mg is contained) through syringe perfusion 33.3% diethylnitrosamine (DEN) suspension 0.0 2 ml(that 1% Tween-80 is newly joined.1 time weekly, totally 15 times.Pathological examination confirms to bring out nasopharyngeal carcinoma success.
(2) get nasopharyngeal carcinoma and bring out successful small white mouse 3 execution, use sampling swab to exist respectively
Test small white mouse pharynx nasalis picks organizes sepage, every mouse 0.5ml.Swab with sepage is stirred mixing in the tumor tissues sepage Sample preservation liquid 2ml that embodiment 2 is obtained, and obtained experimental animal tumor tissues sepage sample liquid 3 parts of normal temperature save backup.
3, the making of clinical detection sample liquid
Sample liquid 1: pathological examination is diagnosed as cervical invasion squamous cell carcinoma.Hold uterine neck sepage sampling thief Via vagina and be slowly inserted into uterus neck, by upper right, bottom right, lower-left, upper left, the wiping in proper order of the uterine neck mouth of pipe, obtain and organize sepage 0.5ml, mix with embodiment 2 gained Sample preservation liquid 2ml, normal temperature saves backup.Sample liquid 2: pathological examination is diagnosed as nodular type nasopharyngeal carcinoma.Hold pharynx nasalis sampler to be inserted by nasal cavity, to go directly nasopharynx along meatus nasi inferior, left and rightly respectively revolve turnback, more in order at posterior wall of nasopharynx, pharyngeal opening of auditory tube, Eustachian tube garden pillow with swallow the wiping of hidden pipe place and obtain and organize sepage 0.5ml, mix with embodiment 2 gained Sample preservation liquid 2ml, normal temperature saves backup.Sample liquid 3: pathological examination is diagnosed as small-cell carcinoma of the lung.Person under inspection please leave and take sputum in specimen bottle, get its 0.5ml and embodiment 2 gained Sample preservation liquid 2ml mixes, normal temperature saves backup.
Sample liquid 4: pathological examination is diagnosed as penis squamous cell carcinoma.Hold sampling thief in the wiping gently of penis coronary sulcus hyperplasia ulcer place, obtain and organize sepage 0.5ml, mix with embodiment 2 gained Sample preservation liquid 2ml, normal temperature saves backup.4, test findings: in table 1 and table 2.
5, statistics display: two groups of P values are all greater than 0.05, difference not statistically significant.Tumour cell hemn content and Sample preservation number of days have nothing to do.
Can be found out by above data; the long-acting tumor tissues sepage Sample preservation liquid that this patent provides; at normal temperatures in Sample preservation 90 days; tumor tissue cell's hemn content is without significant difference; prove that this conserving liquid can protect the stability of tumor tissue cell's hemn effectively; the interference of the various factors existed in sepage can be got rid of, ensure the precision detected.The storage life of sample can reach 90 days at normal temperatures, and therefore, long-acting tumor tissues sepage Sample preservation liquid provided by the invention is a kind of for detecting the fabulous Sample preservation liquid of tumor tissue cell's hemn performance.
Claims (2)
1. a long-acting tumor tissues sepage Sample preservation liquid, it is characterized in that, it is made up of 3.69 ~ 3.76g/L disodium ethylene diamine tetraacetate, 38 ~ 42mg/L isobutyl alcohol, 9.5 ~ 10.5ml/L formaldehyde and 0.1mol/L ~ 1mol/L phosphate buffer; The pH value of described phosphate buffer is 6.2 ~ 7.8.
2. long-acting tumor tissues sepage Sample preservation liquid according to claim 1, is characterized in that, it is that 7.4 phosphate buffers form by the disodium ethylene diamine tetraacetate of 3.72g/L, the isobutyl alcohol of 40mg/L, the formaldehyde of 10ml/L and 0.5mol/L pH value.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110286010A (en) * | 2019-06-06 | 2019-09-27 | 强安医疗器械有限公司 | Liquid sample-specific pretreatment liquid and preparation method thereof in a kind of rectum |
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| CN101999343A (en) * | 2010-10-26 | 2011-04-06 | 深圳华大基因科技有限公司 | Cell preserving fluid and preparation method and use thereof |
| CN102318597A (en) * | 2011-08-19 | 2012-01-18 | 张雪云 | Liquid-based cell preservation liquid composite and preparation method thereof |
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2015
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| EP0511430A2 (en) * | 1991-05-01 | 1992-11-04 | Cytyc Corporation | Cell preservative solution |
| EP0772972A1 (en) * | 1991-05-01 | 1997-05-14 | Cytyc Corporation | Cell preservative solution |
| CN1816634A (en) * | 2003-07-11 | 2006-08-09 | Cytyc公司 | Detection of a target in a preservative solution |
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| CN110286010A (en) * | 2019-06-06 | 2019-09-27 | 强安医疗器械有限公司 | Liquid sample-specific pretreatment liquid and preparation method thereof in a kind of rectum |
| CN110286010B (en) * | 2019-06-06 | 2021-09-24 | 强安医疗器械有限公司 | Special pretreatment liquid for rectal fluid sample and preparation method thereof |
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Application publication date: 20150819 |