CN1967196B - Simplified thin-layer liquid-based cytology cell smeard preparation method - Google Patents
Simplified thin-layer liquid-based cytology cell smeard preparation method Download PDFInfo
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- CN1967196B CN1967196B CN200610021857A CN200610021857A CN1967196B CN 1967196 B CN1967196 B CN 1967196B CN 200610021857 A CN200610021857 A CN 200610021857A CN 200610021857 A CN200610021857 A CN 200610021857A CN 1967196 B CN1967196 B CN 1967196B
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Abstract
The invention discloses a simple thin liquid-based cytology examine method applied for the gynecological examination, the cervical cancer general examine and the cervical cancer preceding lesion of selected crowd, the steps including: using the cervical cells brush and the scraper to collect samples, put samples in the liquid-based cell preservation solution, add base liquid cell processing solution, oscillate, centrifugally separate, take and discard the supernatant samples liquid, using adding samples gun to assimilate sediment samples, smear, sheet drying, using 95% ethanol marinate or spray for fixed, washing and dyeing. The invention has simple method, small steps, high positive detection rate, low misdiagnosis rate, low cost, and it can effectively eliminate the red blood cells, mucus and other interference components from the cervical cytology detection specimens, with good sheet-producing result.
Description
Technical field
The present invention relates to a kind of cytolgical examination method, especially relate to the simplified thin-layer liquid-based cytology detection method that can be used for precancerous lesions of uterine cervix among gynaecology's health check-up, cervical cancer screening and the examination crowd.
Background technology
The whole world has 500,000 cervical carcinoma new cases, China account for 1/4 and be and rise and rejuvenation trend every year.And cervical carcinoma to be human body unique can be by the examination early diagnosis, find precancerous lesion (go up the intracutaneous tumour, intraepilialneoplasia), 100% malignant tumour of curing.Therefore, it is easy to work out a kind of method, with low cost, and accuracy, specificity, the thin-layer liquid-based cytology detection method that susceptibility is high are used for generaI investigation and health check-up, have important clinical meaning and using value.
Traditional common Pap smear inspection has 15%～20% rate of missed diagnosis, because different with flaking method of drawing materials, has 80% cell to be lost
[2], smear inequality, cell accumulation, mucus and erythrocytic influence cause sick cell and pathogen fully not to show, false negative occurs.And the liquid based cytology inspection is that domestic and international at present positive rate of generally acknowledging is the highest, rate of missed diagnosis is minimum, cell smear is best in quality, the pathology composition is preserved inspection method completely.But equipment that this method adopted and reagent, consumptive material are all expensive, and it is difficult to carry out in the general hospital, more is not suitable for generaI investigation in enormous quantities and health check-up.
Summary of the invention
The invention provides a kind of simplified thin-layer liquid-based cytology detection method, it is mainly by adopting liquid based cell preservative fluid and liquid basal cell treating fluid to realize.This method step is few, effective, positive rate is high, rate of missed diagnosis is low, with low cost, be applicable to that vast basic hospital carries out gynaecology's health check-up and cervical cancer screening, reached the purpose of precancerous lesions of uterine cervix among effective examination crowd.
For achieving the above object, the technical solution adopted in the present invention is as follows:
Simplified thin-layer liquid-based cytology detection method is characterized in that comprising the steps:
After A, usefulness cervical canal cytobrush and the scraper plate sampling, place the test tube that is added with liquid based cell preservative fluid to vibrate 1 minute the cytobrush of collecting cell;
B, from the test tube of steps A, take out cytobrush, add base fluid cell treating fluid 4ml vibration 1 minute, place hydro-extractor centrifugal 3～5 minutes again with 500～800 rev/mins;
C, get the supernatant of abandoning in the step B test tube, the vibration several seconds, draw sediment sample (as far as possible avoiding floccus) in the test tube with the application of sample rifle, be applied on the cleaning microslide by same direction, and form thin slice, put in the baking box and to dry in 5 minutes;
D, the thin slice sample after will drying are fixed 5～15 minutes with the alcohol immersion or the spraying of 95% (v/v);
F, the sample with step D after fixing carry out pap staining or HE dyeing after with the tap water washing.
Liquid based cell preservative fluid of the present invention is the mixed liquor of phenol of the glacial acetic acid and 1.91%～2.11% (g/v) of 2% (v/v).
The compound method of described liquid based cell preservative fluid is: after getting 21ml liquefied phenol adding distil water 1280ml and being mixed with the phenol solution that phenol concentration is 1.91%～2.11% (g/v), add the 26ml glacial acetic acid again, promptly getting glacial acetic acid and the concentration that concentration is 2% (v/v) is the phenol mixed liquor of 1.91%～2.11% (g/v).
Liquid basal cell treating fluid of the present invention is the alkaline disodium ethylene diamine tetraacetate of 0.8%～1% (v/v), pH8～8.5.
The compound method of described liquid basal cell treating fluid is: get disodium ethylene diamine tetraacetate (EDTA) 1g and be dissolved in the 100ml distilled water, add 8% sodium hydroxide solution again, the pH of solution is transferred to 8～8.5 promptly.
The formed thin slice of step C of the present invention is the elliptical region of diameter 2cm * 4cm.
The theoretical foundation of simplified thin-layer liquid-based cytology detection method of the present invention:
1, sampling: adopt cervical canal cytobrush and scraper plate, can comprehensive collection uterine neck collar extension glandular scale transitional areas and cervical canal cell.
2, liquid based cell preservative fluid: according to Cervical pathology, physilogical characteristics, take a sample and may contain mucus and red blood cell in this, mucus influence cell and launches and cover sick cell, sheet is read in the interference of red blood cell dilution sick cell.This law is used 1.5%～2% (v/v) glacial acetic acid destruction, lysed erythrocyte, and rapid osmotic is in cell and various pathogen, and the nucleoprotein of pair cell and pathogen plays freezing action, fixed cell nuclear and the various pathogen of deactivation; 1.91%～2.11% (g/v) phenol can be killed pathogen such as mould, bacterium, virus effectively as anticorrosion and bactericidal agent, protection working environment and staff's health.Therefore the present invention preserves liquid and has used a glacial acetic acid and a phenol freezing nuclei albumen dexterously, and the effect that can not solidify cytoplasm protein has avoided epithelial mutual adhesion to pile up, and helps preserving the cell fixation nucleus.Made sample is preserved 2 pericytes does not have the form variation.
3, liquid basal cell treating fluid: cervical mucus is a kind of glycoprotein, has the characteristic of protein, can slowly dissolve in aqueous solution.Use alkaline disodium ethylene diamine tetraacetate can remove iuntercellular and connect, cell dispersion, protein microbeads such as solution fibrin and haemoglobin make that the smear background is Clear ﹠ Transparent, cell disperses, is evenly distributed.Simultaneously, this alkaline solution also plays the acid in the neutralization preservation liquid, and pH is remained on about 7, helps dyeing, guarantees the effect of Color.
Beneficial effect of the present invention shows:
1, adopts cervical canal cytobrush and scraper plate, can accurately get middle sick cell all sidedly at uterine neck collar extension, glandular scale transitional areas and cervical canal position to greatest extent, and fully keep sample, prevent loss.
2, cell fully vibrates through oscillator in the liquid base, and sick cell evenly distributes, grab sample smear, the recall rate of raising sick cell.
3, in the film-making process, used a glacial acetic acid and phenol freezing nuclei albumen dexterously, and the noncondensing effect of pair cell protein can not cause epithelial cell to adhere to accumulation mutually, do not solidify mucus, do not need to do loaded down with trivial details processing, can guarantee that cellular morphology is intact, unchangeability, cell launches good, and mucus and red blood cell can effectively be dispeled, and the smear background clearance is clean, be convenient to easily read sheet, be easy to discern sick cell, thereby improve the positive rate of sick cell, minimizing is failed to pinpoint a disease in diagnosis.
4, the present invention is with low cost, and method is easy, is easy to penetration and promotion, is applicable to that basic hospital carries out cytolgical examination.
Embodiment
Embodiment 1 method step
1, the gynecologist uses scraper plate to scrape sampling originally to wash in the liquid based cell preservative fluid, use the cervical canal cytobrush simultaneously, insert the cervical canal collar extension, take out 7～10 circle backs by turning clockwise, cytobrush inserted in the test tube be added with the 4ml liquid based cell preservative fluid do not take out censorship.Liquid based cytology is preserved liquid: the phenol mixed solution of the glacial acetic acid of 2% (V/V) and 2% (g/V).
2, the test tube that the pathology doctor will contain sample was put on the oscillator vibration 1 minute, fully abandon or adopt cytobrush after the washing, the alkaline disodium ethylene diamine tetra-acetic acid aqueous solution that adds 4ml 1%), vibrated 1 minute, 500～800 rev/mins centrifugal 3～5 minutes, get and abandon supernatant, draw the sediment sample with the application of sample rifle, avoid floccus during absorption as far as possible, be applied on the cleaning microslide by same direction, paint the elliptical region of diameter 2cm * 4cm. promptly paint macroscopic translucent muddy a little thin slice, put oven dry in 5 minutes in the baking box. fix 5～15 minutes with 95% alcohol immersion or spraying again, pap staining or HE dyeing after the washed several times with water from the beginning.
Embodiment 2 contrast tests
Through hundreds of routine clinical researches, its susceptibility is 91.6%, and negative predictive value reaches 98.7%.Positive rate is higher than the cytolgical examination that common Pap smear (p＜0.05) therefore is applicable to gynaecology in enormous quantities generaI investigation, health check-up and basic hospital, and its cheap charge can reduce the patient burden.See Table 1, table 2.
Routine simple and easy liquid based cytology check result of table 1209 and common Pap smear check result are relatively
| ASCUS ％ | LSIL ％ | HSIL ％ | Pathogen+% | |
| The film-making of liquid base | 13 examples 6.2 | 6 examples 2.8 | 4 examples 1.9 | 5 examples 2.3 |
| Tradition Pasteur | 8 examples 3.8 | 4 examples 1.9 | 3 examples 1.4 | 0 example |
Table 2 smear mass ratio
| Quantitative target | Example % | + routine % | ++ routine % | +++routine % |
| Red cell volume | 209 100 | |||
| Cell degeneration | 209 100 | |||
| The mucus background | 192 91.8 | 12 5.7 | 5 2.3 | |
| Cell accumulation | 188 89.9 | 16 7.6 | 5 2.3 |
Claims (2)
1. the simplified thin-layer liquid basal cell is learned the cell smear preparation method, it is characterized in that comprising the steps:
After A, usefulness cervical canal cytobrush and the scraper plate sampling, place the test tube that is added with liquid based cell preservative fluid to vibrate 1 minute the cytobrush of collecting cell;
B, from the test tube of steps A, take out cytobrush, add base fluid cell treating fluid vibration 1 minute, place hydro-extractor centrifugal 3～5 minutes again with 500～800 rev/mins;
C, get the supernatant of abandoning in the step B test tube, draw sediment sample in the test tube, be applied on the cleaning microslide by same direction, and form thin slice, put oven dry in 5 minutes in the baking box with the application of sample rifle;
D, the thin slice sample volume fraction after will drying are that 95% alcohol immersion or spraying are fixed 5～15 minutes;
E, the sample with step D after fixing carry out pap staining or HE dyeing after with the tap water washing;
Wherein, the described liquid based cell preservative fluid of steps A is that the mass/volume mark is that 2% glacial acetic acid and mass/volume mark are the mixed liquor of 1.91%～2.11% phenol; The described liquid basal cell treating fluid of step B is that volume fraction is 0.8%～1%, the alkaline disodium ethylene diamine tetraacetate of pH8～8.5.
2. simplified thin-layer liquid basal cell according to claim 1 is learned the cell smear preparation method, the compound method that it is characterized in that described liquid based cell preservative fluid is: getting 21ml liquefied phenol adding distil water 1280ml, to be mixed with the mass/volume mark be after 1.91%～2.11% the phenol solution, add the 26ml glacial acetic acid again, promptly get volume fraction and be 2% glacial acetic acid and mass/volume mark and be 1.91%～2.11% phenol mixed liquor.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022149974A1 (en) | 2021-01-05 | 2022-07-14 | Cheong Mun Shun | Automated slide preparing system and method thereof |
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| WO2010134073A1 (en) * | 2009-05-19 | 2010-11-25 | Zetiq Technologies Ltd. | Kits for and methods of differential staining of cervical cancer cells and/or tissues |
| CN102749230A (en) * | 2011-04-22 | 2012-10-24 | 麦克奥迪（厦门）医疗诊断系统有限公司 | Manual semi-quantitative liquid-base cytological flaking method |
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| CN102393318A (en) * | 2011-08-19 | 2012-03-28 | 张雪云 | Simple and quick preparation method of thin-layer liquid-based cytology cell smear |
| CN102944461A (en) * | 2012-11-17 | 2013-02-27 | 武汉市康创科技有限公司 | Diluent applied to liquid-based filmmaking and preparation and using method thereof |
| CN103900864A (en) * | 2012-12-25 | 2014-07-02 | 泰州医药城博奥邦科生物科技有限公司 | Exfoliated cell chip |
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| CN103471892A (en) * | 2013-09-22 | 2013-12-25 | 厦门大学附属第一医院 | Method for preparing pulled type cervical smear |
| CN103875653B (en) * | 2014-02-11 | 2015-03-18 | 中国人民解放军北京军区总医院 | Endocervical cell preserving fluid and method for preparing endocervical cell specimen |
| CN104614526A (en) * | 2015-01-08 | 2015-05-13 | 杭州泰申生物技术有限公司 | Squamous cell carcinoma SCC-TCT combined detection method |
| CN105675374A (en) * | 2016-03-10 | 2016-06-15 | 刘崇梅 | Semi-automatic smearing method of hydrothorax and ascite liquid-base cytology |
| CN106442078B (en) * | 2016-08-25 | 2019-09-10 | 四川金域医学检验中心有限公司 | A kind of alkalinity liquid basal cell saves treatment fluid and preparation method thereof |
| CN107389392A (en) * | 2017-05-25 | 2017-11-24 | 长沙金域医学检验所有限公司 | A kind of cytologic slide detection method |
| CN107389415A (en) * | 2017-05-25 | 2017-11-24 | 长沙金域医学检验所有限公司 | A kind of Liquid based cytology test method |
| CN109490305A (en) * | 2018-12-12 | 2019-03-19 | 西安医学院 | A kind of cell smear detection method |
| CN112179735A (en) * | 2020-09-29 | 2021-01-05 | 江苏硕世生物科技股份有限公司 | Preparation method of cell line immunohistochemical reference substance |
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