CN106645678A - Detection method of oral swab - Google Patents
Detection method of oral swab Download PDFInfo
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- CN106645678A CN106645678A CN201611188914.0A CN201611188914A CN106645678A CN 106645678 A CN106645678 A CN 106645678A CN 201611188914 A CN201611188914 A CN 201611188914A CN 106645678 A CN106645678 A CN 106645678A
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- elisa plate
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- Urology & Nephrology (AREA)
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Abstract
The invention relates to a detection method of an oral swab. The detection method comprises nine steps, so that a detection process of the oral swab is divided in detail. The detection method has the beneficial effects that the operation is simple, comprehensiveness in consideration is realized, and a storage temperature range and a storage time duration are controlled in the detection of the oral swab, so that the oral swab can be better applied to a practical situation.
Description
Technical field.
The present invention relates to oral medical field, is related to a kind of detection method of buccal swab.
Background technology.
Buccal swab is nucleic acid source important in the research of molecular epidemiology, science of heredity and medical jurisprudence, and its collection has
Sampling simplicity, safety, low cost, AT advantage.Low temperature detects that the cold strike to nucleic acid makes it to obtain after being saved
Preferable Detection results, high temperature detection nucleic acid accelerates again the reproduction speed of energetic supersession aggravation, bacterium, cause nucleic acid short
The decline phenomena of mortality are just showed in time.Show that the optimal storage temperature of buccal swab nucleic acid, must at 25 DEG C -45 DEG C through research
Must detect at this temperature, and detection method effect on driving birds is not good now, accordingly, it would be desirable to a kind of detection method can meet normal temperature it is long when
Between purpose.
The content of the invention.
In view of this, the present invention provides a kind of a kind of detection side of buccal swab that solution or part solve the above problems
Method.
To reach the effect of above-mentioned technical proposal, the technical scheme is that：A kind of detection method of buccal swab, its
It is characterised by, comprises the steps of：
50 parts of buccal swabs are taken, KJ001～KJ0050 is numbered, concrete detection method is as follows：
All buccal swabs are placed in before detection should be recovered to 25-27 DEG C of room temperature, and lightly rotation and
Concussion is mixed；
1) the taking-up coated ELISA Plate of antibody from hermetic bag, marks the position of sample in recording sheet；
2) each negative control for adding 80 μ l not diluted in A2 holes and B2 holes；
3) the not diluted positive controls of 80 μ l are respectively added in C2 holes and D2 holes；
4) 80 μ l test samples are separately added in remaining hole；
5) cap covers ELISA Plate is used, is incubated 40 minutes at 25-27 DEG C of room temperature；
6) content in (pouring out) hole is suctioned out, with ultrapure washing 3-5 time, the ultrapure water capacity of 350 μ l is added per hole, most
Afterwards a ultra-pure water is washed while using blotting paper, and gently pat；
7) 80 μ l ELIAS secondary antibodies are added in each hole of the coated ELISA Plate of the antibody, with cap covers, in the 25- of room temperature
It is incubated 80 minutes at 27 DEG C, plus 100 μ l substrate reagents are in each hole of the coated ELISA Plate of the antibody, with cap covers,
It is incubated 80 minutes at 25-27 DEG C of room temperature；
8) 100 μ l terminate liquids are added in each hole of the coated ELISA Plate of the antibody, and terminating reaction.
The invention has the beneficial effects as follows：Advantages of the present invention is comprehensive for simple to operate, consideration, in the detection of buccal swab
In, the temperature range of preservation and the time span of preservation being controlled, can preferably be applied in actual conditions.
Specific embodiment.
In order that the technical problem to be solved, technical scheme and beneficial effect become more apparent, below tie
Embodiment is closed, the present invention will be described in detail.It should be noted that specific embodiment described herein is only to explain
The present invention, is not intended to limit the present invention, and the product that can realize said function belongs to equivalent and improvement, is all contained in this
Within bright protection domain.Concrete grammar is as follows：
Dilution is the key factor for affecting buccal swab nucleic acid normal temperature preservation effect, and the Main Function of dilution is：One
Aspect is used to expand nucleic acid volume to meet Production requirement；The opposing party and protect the functional character of nucleic acid.It is contained in dilution
Protective effect of the various composition to buccal swab nucleic acid is also different, nucleic acid need produce energy with maintain cell metabolism and from
Body is moved.Glucose can not only provide energy for nucleic acid, can also maintain the osmotic pressure of dilution, make in most formulas
With.Peracid crosses the environment of alkali and is all unfavorable for that nucleic acid is survived, and needs to add a certain amount of buffer substance to maintain nucleic acid in dilution
pH.Sodium acid carbonate or sodium citrate are often used to the buffer capacity of the buffers such as the pH, Tris and HEPES of regulation dilution more
By force.EDTA
A kind of chelating agent, can especially Ca2+ is combined with the metal ion in dilution, reduce enter it is intracellular
Ca2+, prevents from inducing nucleic acid capacitation and the generation of acrosome reaction, and to reach nucleic acid activity is suppressed, and increases the effect of nucleic acid preservation time
Really.It is certain density+Na+-K+ pumps operating on nucleic acid film can be maintained, but must assure that the K+ concentration in cell nucleic acid is higher than cell
Outer K concentration, could so produce ATP and maintain depositing and moving for nucleic acid, thus dilution can add appropriate KC1.In addition, dilute
Releasing addition antibiotic in liquid can suppress the breeding of bacterium, main conventional antibiosis to have penicillin, gentamicin etc..But by
In modern society for the abuse of antibiotic so that many bacteriums obvious resistance for common antibiotics have, so, it is new
Antibiotic be applied to buccal swab nucleic acid normal temperature preserve research it is extremely urgent.
Buccal swab nucleic acid is individually preserved, and harmful microorganism therein can not only fight for nutriment, and metabolism is produced
Raw toxic metabolite waste can change Conservation environment, have a strong impact on preservation effect.It is existing mushroom have Escherichia coli, Salmonella
Bacterium, klebsiella, proteus and corynebacteria etc..Gram-negative bacteria is mushroom common in boar semen.Therefore dilute
Release to add in agent has the antibiotic of inhibition optimal to Gram-negative bacteria.Add gentamicin in dilution to suppress thin
The growth of bacterium, the ill-effect to cell is little.Adding the PVP-I of 0.2mg/ml in dilution can kill malignant bacteria,
And quality index is preserved on semen at normal temperature without impact.There is researcher to carry out bacteria distribution, mirror in the buccal swab to separate sources
Find when fixed, culture and drug sensitive test, in five kinds of bacteriums of detection, the Kui Ruo ketone such as Ciprofloxacin, Ofloxacin and Norfloxacin
Class medicine is preferable to its inhibition, and finds that proteus, corynebacteria and salmonella have to penicillin and clindamycin
There is drug resistance.The fungistatic effect of antibiotic and combination to bacterium in Buffalo Semen is compared, is as a result shown, penicillin, streptomysin
With gentamicin all with antibacterial circle, and tylosin, cillimycin, neomycin and spectinomycin have different degrees of resistance
Phenomenon.The big antibiotic combinations of safe woods are celebrated in terms of antibiotic combinations optimal.In practical application, the selection of antibiotic can be because of dilution
Agent composition it is different and different.
The preferred embodiments of the invention is the foregoing is only, the claims of the present invention are not limited to.
It is simultaneously described above, for those skilled in the technology concerned it would be appreciated that and implement, therefore other are based on institute of the present invention
The equivalent that disclosure is completed changes, and should be included in the covering scope of the claims.
The invention has the beneficial effects as follows：Advantages of the present invention is comprehensive for simple to operate, consideration, in the detection of buccal swab
In, the temperature range of preservation and the time span of preservation being controlled, can preferably be applied in actual conditions.
Claims (1)
1. a kind of detection method of buccal swab, it is characterised in that comprise the steps of：
50 parts of buccal swabs are taken, KJ001～KJ0050 is numbered, concrete detection method is as follows：
All buccal swabs are placed in before detection should be recovered to 25-27 DEG C of room temperature, and lightly rotated and shaken
Mixed；
1) the taking-up coated ELISA Plate of antibody from hermetic bag, marks the position of sample in recording sheet；
2) each negative control for adding 80 μ l not diluted in A2 holes and B2 holes；
3) the not diluted positive controls of 80 μ l are respectively added in C2 holes and D2 holes；
4) 80 μ l test samples are separately added in remaining hole；
5) cap covers ELISA Plate is used, is incubated 40 minutes at 25-27 DEG C of room temperature；
6) content in (pouring out) hole is suctioned out, with ultrapure washing 3-5 time, the ultrapure water capacity of 350 μ l is added per hole, last
Secondary ultrapure washing uses blotting paper simultaneously, and gently pats；
7) 80 μ l ELIAS secondary antibodies are added in each hole of the coated ELISA Plate of the antibody, with cap covers, at 25-27 DEG C of room temperature
Lower incubation 80 minutes, plus 100 μ l substrate reagents are in each hole of the coated ELISA Plate of the antibody, with cap covers, in room temperature
25-27 DEG C at be incubated 80 minutes；
8) 100 μ l terminate liquids are added in each hole of the coated ELISA Plate of the antibody, and terminating reaction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611188914.0A CN106645678A (en) | 2016-12-21 | 2016-12-21 | Detection method of oral swab |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611188914.0A CN106645678A (en) | 2016-12-21 | 2016-12-21 | Detection method of oral swab |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106645678A true CN106645678A (en) | 2017-05-10 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611188914.0A Pending CN106645678A (en) | 2016-12-21 | 2016-12-21 | Detection method of oral swab |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604922A (en) * | 2013-11-20 | 2014-02-26 | 天津市宝坻区人民医院 | Respiratory syncytial virus detection kit and use method thereof |
| WO2016053689A1 (en) * | 2014-09-30 | 2016-04-07 | Ge Healthcare Uk Limited | Methods and devices relating to the detection of oral cancer biomarkers |
| CN106244582A (en) * | 2016-08-30 | 2016-12-21 | 苏州凡济医疗器械有限公司 | A kind of buccal swab nucleic acid room temperature store method |
-
2016
- 2016-12-21 CN CN201611188914.0A patent/CN106645678A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604922A (en) * | 2013-11-20 | 2014-02-26 | 天津市宝坻区人民医院 | Respiratory syncytial virus detection kit and use method thereof |
| WO2016053689A1 (en) * | 2014-09-30 | 2016-04-07 | Ge Healthcare Uk Limited | Methods and devices relating to the detection of oral cancer biomarkers |
| CN106244582A (en) * | 2016-08-30 | 2016-12-21 | 苏州凡济医疗器械有限公司 | A kind of buccal swab nucleic acid room temperature store method |
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Application publication date: 20170510 |
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