CN105274185A - Sputum treating liquid for detecting Mycobacterium tuberculosis - Google Patents
Sputum treating liquid for detecting Mycobacterium tuberculosis Download PDFInfo
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- CN105274185A CN105274185A CN201510784562.4A CN201510784562A CN105274185A CN 105274185 A CN105274185 A CN 105274185A CN 201510784562 A CN201510784562 A CN 201510784562A CN 105274185 A CN105274185 A CN 105274185A
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- sputum
- treatment solution
- tuberculosis
- treating liquid
- tubercule bacillus
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Abstract
A sputum treating liquid for inspecting standard films of Mycobacterium tuberculosis in sputum comprises ethyl alcohol, isopropyl alcohol, boric acid, disodium EDTA, SDS, a mucus dissolving agent and distilled water and is capable of dissolving phlegm, deactivating pathogenic bacteria such as Mycobacterium tuberculosis as well as viruses in sputum, preventing corrosion and splitting exfoliative cells in sputum. Applied with simple centrifugal operation, the sputum treatment with the sputum treating liquid enables 8 to 10 spits of sputum to be treated at a time, and even sputum left for 24 hours, the treatment quantity of sputum is increased, standard operation is achieved through a standard automatic film-making technique, the operation is simple and easy, detection sensitivity is improved, detection is accurate and reliable, and the sputum treating liquid is suitable for the general investigation of tuberculosis high-risk groups, screening of rural tuberculosis and clinical diagnosis of tuberculosis.
Description
Technical field
The present invention relates to a kind of sputum treatment solution detected for tubercule bacillus, for pulmonary tuberculosis high risk population generaI investigation, Pulmonary Tuberculosis in Rural examination and clinical diagnosis of tuberculosis, belong to external diagnosis reagent field.
Background technology
Mycobacterium tuberculosis (M.tuberculosis), is commonly called as tubercule bacillus, is cause pathogenic bacteria lungy.The each organ of whole body can be invaded, but be the most common with pulmonary tuberculosis.Tuberculosis is still important transmissible disease so far.According to WHO, about there are 8,000,000 new cases to occur every year, have at least 3,000,000 people to die from this disease.China is one of 22 tuberculosis states occurred frequently in the whole world, and the tuberculosis patient of West Pacific region 70% is in China.According to Ministry of Health's investigation display, the whole nation about has 5.5 hundred million people to infect tubercule bacillus, existing active tuberculosis patient 4,500,000, wherein has communicable patient 1,500,000, annual newly-increased tuberculosis patient about 1,300,000.The patient of China 80% is distributed in rural area, and the patient of 75% is between twenty and fifty.China dies from number lungy for 1 year and reaches 130,000, suitable with the number dying from traffic accident, occupy first of various disease death reason, living standards of the people improve after the founding of the state, hygienic state is improved, particularly carried out mass prevention and mass treatment, the general bcg vaccination of children, M & M lungy greatly reduces.But it should be noted that some area is in the world because of the application of acquired immune deficiency syndrome (AIDS), drug abuse, immunosuppressor, the reason such as excessive drinking and poverty, sickness rate is on the rise again.Mycobacterium tuberculosis is elongated slightly bending bacillus, size 1 ~ 4 × 0.4 μm.The bacteria cell wall lipid content of Mycobacterium is higher, accounts for 60% of dry weight, particularly has a large amount of mycolic acid (mycolicacid) to be enclosed in the outside of peptidoglycan layer, can affect penetrating of dyestuff.Containing lipid in Mycobacterium cell wall, therefore to alcohol sensible, in 70% ethanol, 2min is dead.In addition, lipid can prevent thalline loss of moist, therefore strong especially to the resistibility of drying.Stick on dust and keep infectivity 8 ~ 10 days, can survive in dry phlegm 6 ~ 8 months.Mycobacterium tuberculosis, to damp and hot sensitivity, heats 62 ~ 63 DEG C in a liquid and within 3 minutes, is namely killed for 15 minutes or 70 DEG C.Mycobacterium tuberculosis is to ultraviolet-sensitive.Direct sun exposure a few hours can be killed, and can be used for the sterilization of tubercular's clothes, books etc.Mycobacterium generally uses neat Buddhist nun (Ziehl-Neelsen) Ziehi-Neelsen stain, heats after dyeing and can catch, but not easily to decolour with 3% acidic alcohol with 5% carbolfuchsin.Redye with methylene blue if add, then mycobacterium takes on a red color again, and the material in other bacteriums and background is blue.The detection of tubercule bacillus is the key reducing M & M lungy with prevention.The method of current detection tubercule bacillus mainly contains etiology method, immunological method and molecular biology method.
Pathogeny detection is tuberculosis diagnosis method unique at present, and etiology method mainly contains two kinds: one, Sputum smears method.This is the most basic bacteriology checking method that tuberculosis laboratory, the whole world is all using, and have fast and convenient and inexpensive etc. advantage, smear methods has following several: 1, Ziehi-Neelsen stain.Classical way, clinically widely uses.2, fluorescent method.With fluorescent microscope, can increase work efficiency.3, dye method is joined.Be combined with fluorescence colour by Ziehi-Neelsen stain.Sputum smears method is the clinical diagnosis method that cheapness is easy fast, but traditional Sputum smears method positive rate is too low.Cannot stdn, the technology of examined person and sense of responsibility impact larger.Two, Sputum culturing method.The advantage of tubercule bacillus general culture method is that sensitivity, specific degree, accuracy are higher, and be that qualification is dead, the reliable method of viable bacteria, being the irreplaceable method of current living stems, is also the main monitoring index of chemotherapy side effect; Shortcoming is that incubation time is long, needs 4-8 week just can report the result, the shell vial method newly gone out, and instrument price is expensive and need inlet liquid substratum, costly, and unfavorable popularization.
Sputum smears detection mode clinical is at present after obtaining sputum sample, get wherein a part of direct smear dyeing microscopic examination, the cell come off because sputum sample viscosity material is many, mobility is bad, containing human body and the tubercule bacillus limited amount that may exist, all bacteriums in sputum cannot effectively be collected on slide by this way.Therefore, if take corresponding way or reagent sputum to be carried out liquefaction processing release tubercule bacillus, simultaneously cracking human body cast-off cells wherein, so just by simple centrifugal treating whole tubercule bacillus can be collected and carry out film-making.Not only increase the bacterium visual field density under microscope, also eliminate the interference of mucus and human body cast-off cells simultaneously, greatly improve diagnostic sensitivity.
Summary of the invention
In order to solve the shortcoming in Sputum smears practical application, contriver, by large quantifier elimination and screening experiment, obtains the sputum prescription for the treatment of liquid of innovation, and this treatment solution has use safety, not containing hazardous chemical; Biological safety is good, pathogenic micro-organism in the process of liquefaction sputum in quick inactivating sample; Collection bacterium is effective, can a large amount of sputum of primary treatment, reaches the object of concentrated bacterium; Easy to operate, the advantages such as normal-temperature operation, normal temperature transport.
Utilize the sputum treatment solution process sputum in this invention, in simple blending process, sputum can be clarified very soon, cause of disease in sputum sample obtains thorough deactivation simultaneously cracking human body cast-off cells wherein and keeps the form of tubercule bacillus complete, and after film-making, the acid-fast stain in the tubercule bacillus that slide adheres to and downstream, fluorescent dye or combined staining operate mates completely.
Embodiment
Specific embodiments of the invention following (preparation 100ml treatment solution):
1. get distilled water 65 milliliters, put into measuring cup, measure boric acid 0.6g, EDETATE SODIUM 0.1g, SDS0.1g respectively, add in measuring cup to stir and fully dissolve for 10 minutes.
2. get dehydrated alcohol 10 milliliters respectively, Virahol 20 milliliters, mucolytic agent 5 milliliters add in measuring cup to stir and be mixed for 5 minutes.
3. step one and step 2 liquid are poured into respectively in beaker to stir and be mixed for 10 minutes.Namely the configuration of sputum treatment solution is complete.
using method
To be spat into by sputum in the pipe that sputum treatment solution is housed or get sputum add in treatment solution from getting phlegm tank, mixing allows treatment solution carry out liquefaction processing to sample, leaves standstill the 1-3 minute that vibrates after 5 minutes, marker samples; Heat sample, put into 70 DEG C of constant temperature water box 10-15 minute, this step can be omitted, because sputum treatment solution is by the tubercule bacillus complete inactivation in sputum, this step is the further guarantee to deactivation; Taking-up sample is put into whizzer directly centrifugal (4000 turns) 5 minutes, remove supernatant, precipitate and suspend for subsequent use with a small amount of pure water.
Actual use shows, after using this treatment solution, remain under mirror without mucus, cast-off cells interference is few, clear background, bacterium visual field density is large, and ne ar is intact, completely eliminated the background interference of mucus, add bacterial density, improve sensitivity, reduce difficulty and the fatigue strength of read tablet person, improve the efficiency of diagnosis.
technical superiority
The present invention has abandoned traditional strong alkali solution or the sputum treatment solution containing toxic starting materials, pass through formula optimization, obtain and be simple and easy to use, the sputum treatment solution of safety non-toxic, and by utilizing treatment solution to carry out inactivation treatment to cause of disease in sample phase, improve biological safety, simultaneously by the method for osmotic pressure, human body cast-off cells in cracking sputum, reduce the interference of background.
The present invention is easy to use, and sputum treatment solution normal temperature uses, normal temperature transport is preserved.
The present invention is with low cost, preparation is simple, to auxiliary appliance without particular requirement, does not need to carry out loaded down with trivial details training to user, is applicable to extensive tuberculosis Screening Diagnosis.
Claims (6)
1., for the sputum treatment solution that tubercule bacillus detects, it is characterized in that it is made up of following composition and part by weight: ethanol 5-20%, Virahol 10-25%, boric acid 0.2-1.5%, EDETATE SODIUM 0.01-0.2%, SDS0.001-0.1%, mucolytic agent 3-12%, distilled water 40-75%.
2. sputum treatment solution according to claim 1, is characterized in that not needing concuss in sputum treating processes or rocking, and only needs to mix gently and can melt sputum.
3. sputum treatment solution according to claim 1, is characterized in that, in the process sampled, cause of disease contained in sputum comprises tubercule bacillus and was namely inactivated within 1 minute, avoids infected possibility in operator's subsequent processes.
4. sputum treatment solution according to claim 1, the amount of process sputum improves a lot, and maximum ratio can reach 2.5:1(sputum treatment solution: sputum).
5. sputum treatment solution according to claim 1, it is characterized in that sputum treatment solution can thoroughly liquefy sputum, cracking human body cast-off cells, release tubercule bacillus, in conjunction with simple centrifugally operated, get final product enrichment of bacterial, reach and put forward highly sensitive object, simultaneously for the cracking of human body cast-off cells, achieve the object reducing cellular context.
6. sputum process according to claim 1, can use at 4-70 DEG C and operate, also can transport at-10-40 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201510784562.4A CN105274185A (en) | 2015-11-16 | 2015-11-16 | Sputum treating liquid for detecting Mycobacterium tuberculosis |
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| CN201510784562.4A CN105274185A (en) | 2015-11-16 | 2015-11-16 | Sputum treating liquid for detecting Mycobacterium tuberculosis |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967778A (en) * | 2017-03-27 | 2017-07-21 | 南京邦博生物技术有限公司 | A kind of quick sputum dissolving preserves liquid |
| CN107400695A (en) * | 2017-03-06 | 2017-11-28 | 李齐 | A kind of sputum microorganism sample homogeneity reagent |
| CN112574938A (en) * | 2019-09-29 | 2021-03-30 | 广东体必康生物科技有限公司 | Sputum treatment fluid for membrane filtration enriched bacteria and application thereof |
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| CN1816634A (en) * | 2003-07-11 | 2006-08-09 | Cytyc公司 | Detection of a target in a preservative solution |
| WO2010088191A1 (en) * | 2009-01-27 | 2010-08-05 | Teleflex Medical Incorporated | Sputum dissolving suctioning solution for endotracheal and tracheostomy tubes |
| CN102226160A (en) * | 2011-05-13 | 2011-10-26 | 广州金域医学检验中心有限公司 | Sputasol for isolated culture of Legionnella |
| CN102258003A (en) * | 2010-05-28 | 2011-11-30 | 孝感市中心医院 | Liquid based cell preserving fluid |
| CN102318597A (en) * | 2011-08-19 | 2012-01-18 | 张雪云 | Liquid-based cell preservation liquid composite and preparation method thereof |
| CN104561230A (en) * | 2015-01-22 | 2015-04-29 | 四川金域医学检验中心有限公司 | Sputum specimen homogenization method and preparation of reagent for medical detection of sputum specimen |
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2015
- 2015-11-16 CN CN201510784562.4A patent/CN105274185A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1816634A (en) * | 2003-07-11 | 2006-08-09 | Cytyc公司 | Detection of a target in a preservative solution |
| WO2010088191A1 (en) * | 2009-01-27 | 2010-08-05 | Teleflex Medical Incorporated | Sputum dissolving suctioning solution for endotracheal and tracheostomy tubes |
| CN102258003A (en) * | 2010-05-28 | 2011-11-30 | 孝感市中心医院 | Liquid based cell preserving fluid |
| CN102226160A (en) * | 2011-05-13 | 2011-10-26 | 广州金域医学检验中心有限公司 | Sputasol for isolated culture of Legionnella |
| CN102318597A (en) * | 2011-08-19 | 2012-01-18 | 张雪云 | Liquid-based cell preservation liquid composite and preparation method thereof |
| CN104561230A (en) * | 2015-01-22 | 2015-04-29 | 四川金域医学检验中心有限公司 | Sputum specimen homogenization method and preparation of reagent for medical detection of sputum specimen |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107400695A (en) * | 2017-03-06 | 2017-11-28 | 李齐 | A kind of sputum microorganism sample homogeneity reagent |
| CN107400695B (en) * | 2017-03-06 | 2018-03-06 | 李齐 | A kind of sputum sample homogeneity method and the reagent used |
| CN106967778A (en) * | 2017-03-27 | 2017-07-21 | 南京邦博生物技术有限公司 | A kind of quick sputum dissolving preserves liquid |
| CN106967778B (en) * | 2017-03-27 | 2018-04-06 | 南京邦博生物技术有限公司 | A kind of quick sputum dissolving preserves liquid |
| CN112574938A (en) * | 2019-09-29 | 2021-03-30 | 广东体必康生物科技有限公司 | Sputum treatment fluid for membrane filtration enriched bacteria and application thereof |
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Application publication date: 20160127 |