CN106234354A - A kind of cervix cell preservation solution - Google Patents
A kind of cervix cell preservation solution Download PDFInfo
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- CN106234354A CN106234354A CN201610875401.0A CN201610875401A CN106234354A CN 106234354 A CN106234354 A CN 106234354A CN 201610875401 A CN201610875401 A CN 201610875401A CN 106234354 A CN106234354 A CN 106234354A
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- liquid
- ammonium chloride
- preservation
- trimethyl ammonium
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
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- Environmental Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of cervix cell preservation solution, including PBS, Dodecyl trimethyl ammonium chloride and E.C. 3.4.21.64, described Dodecyl trimethyl ammonium chloride is first mixed in PBS to be formed and preserves liquid for the first time, and described Dodecyl trimethyl ammonium chloride is 5 15% at the first mass percent concentration preserved in liquid；Being mixed into described first preservation liquid after described E.C. 3.4.21.64 and form finished product preservation liquid, described E.C. 3.4.21.64 is 4 6mg/ml with the mass volume ratio concentration of described first preservation liquid.This cervix cell preservation solution has easy and simple to handle, preparation easily, to the treatment effect of courageous and upright sample and mucus sample preferably and advantage that suppression will not occur.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of cell-preservation liquid, particularly relate to a kind of cervix cell preservation solution.
Background technology
In the detection of existing HPV (human papillomavirus) typing, sample process is a key link, directly determines final
The accuracy of result.Current cervix cell preservation solution is normal saline, and the sample process of this cell-preservation liquid exists following institute
The interference factor stated:
1, blood sample, the sample carrying out there are about in the sample of HPV detection 1/3rd is courage and uprightness, and erythrocyte is extracting
Being mainly composed of hemoglobin etc. after rupturing during DNA, hemoglobin cannot thoroughly be removed, and directly affects follow-up
PCR (polymerase chain reaction) process, produces inhibitory action to this process, causes result false negative.
2, mucus sample, the sample carrying out there are about in the sample of HPV detection half contains mucus.The existence of mucus affects sample
This suction process, causes suction sample not enough, causes false negative.
To this end, existing cervix cell preservation solution just can carry out HPV detection after needing first to carry out eluting process so that process
Process becomes loaded down with trivial details, once skips eluting and processes, easily produces erroneous judgement thus have a negative impact patient health.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the PBS cell-preservation liquid of prior art to need first to wash
The deficiency of HPV detection just can be carried out, it is provided that a kind of cervix cell preservation solution after de-process.
For solving above-mentioned technical problem, the present invention adopts the following technical scheme that:
A kind of cervix cell preservation solution, it is characterised in that: include PBS, Dodecyl trimethyl ammonium chloride and egg
White enzyme K, described Dodecyl trimethyl ammonium chloride is first mixed in PBS to be formed and preserves liquid for the first time, described dodecyl three
Ammonio methacrylate is 5-15% at the first mass percent concentration preserved in liquid；Described first guarantor it is mixed into after described E.C. 3.4.21.64
Liquid storage forms finished product and preserves liquid, and described E.C. 3.4.21.64 is 4-6mg/ml with the mass volume ratio concentration of described first preservation liquid.
Described PBS is the phosphate buffer (being called for short PBS-T) of the pH=7.4 containing 0.05% tween 20.
Above-mentioned cervix cell preservation solution is used to have the advantage that
1, the trimethyl component in the cervix cell preservation solution of the present invention can be destroyed erythrocyte and play haemolysis
Effect, E.C. 3.4.21.64 can effectively decompose mucus, and PBS buffer agent can play the effect preserving cervix cells.
2, the cervix cell preservation solution of the present invention just can directly detect without carrying out eluting process, and detects essence
Accuracy of detection after degree and existing cervix cell preservation solution eluting is suitable, simplifies operating procedure, improves the detection of HPV virus
Efficiency.
3, the cervix cell preservation solution raw material of the present invention is cheap and easily-available, easily prepared.
Accompanying drawing explanation
Fig. 1 is HPV.16+PCR expands standard curve, and abscissa is period, and vertical coordinate is copy number.
Fig. 2 is to use producer's cell-preservation liquid and cell-preservation liquid of the present invention to HPV16 respectively+Carry out preserving and extract amplification
The amplification curve obtained, abscissa is period, and vertical coordinate is copy number.
Fig. 3 is to use producer's cell-preservation liquid and cell-preservation liquid of the present invention to HPV18 respectively+Carry out preserving and extract amplification
The amplification curve obtained, abscissa is period, and vertical coordinate is copy number.
Fig. 4 is to use producer's cell-preservation liquid and cell-preservation liquid of the present invention to HPV31 respectively+Carry out preserving and extract amplification
The amplification curve obtained, abscissa is period, and vertical coordinate is copy number.
Fig. 5 is to use producer's cell-preservation liquid and cell-preservation liquid of the present invention to HPV58 respectively+Carry out preserving and extract amplification
The amplification curve obtained, abscissa is period, and vertical coordinate is copy number.
Fig. 6 extracts for using producer's cell-preservation liquid and cell-preservation liquid of the present invention to carry out HPVCP8304 preserving respectively
The amplification curve that amplification obtains, abscissa is period, and vertical coordinate is copy number.
Fig. 7 shows and verifies, by flow hybridization method checking type qualification test, the effect that the present invention is played.
Detailed description of the invention
Experimental example 1
Test in cervix cell preservation solution the mass fraction of Dodecyl trimethyl ammonium chloride to erythroclasis effect
With the impact on HPV testing result:
Experimentation is as follows: is mixed in PBS to be formed by Dodecyl trimethyl ammonium chloride and preserves liquid for the first time；PBS
Buffer is the phosphate buffer of the pH=7.4 containing 0.05% tween 20.
The first liquid that preserves of the Dodecyl trimethyl ammonium chloride containing different quality percentage ratio is respectively placed in different anti-
Answer in container and clear marking, separately sampled from same cervix cells sample put in respective reaction vessel, at identical bar
Reaction result such as following table is recorded after reaction under part；
Table 1:
Reaction result is as shown in table 1: when Dodecyl trimethyl ammonium chloride in the first mass percent preserved in liquid is
When 5%, hemolyzing effect is preferable, and the testing result unrestraint phenomenon to HPV, when Dodecyl trimethyl ammonium chloride is for the first time
Preserve and when the mass percent in liquid is 10%, 15%, can accomplish that complete hemolysis and the testing result unrestraint to HPV are existing
As, when Dodecyl trimethyl ammonium chloride can be accomplished completely when the first mass percent preserved in liquid is 20%, 25%
Haemolysis, but the testing result of HPV is had part suppression, when in preservation liquid without trimethyl, entirely without
Haemolysis and completely inhibit the testing result of HPV, to sum up, the mass percent of preferred Dodecyl trimethyl ammonium chloride is
5%-15%, the mass percent of most preferred Dodecyl trimethyl ammonium chloride is 10%.
Experimental example 2
Tested same volume containing mass percent be 10% Dodecyl trimethyl ammonium chloride preserve liquid for the first time
In, it being separately added into the E.C. 3.4.21.64 of different quality, the finished product forming different quality volume by volume concentration preserves liquid to the process to mucus
Effect and the impact on HPV testing result.
Experimentation is as follows: is at the beginning of 10% Dodecyl trimethyl ammonium chloride in same volume containing mass percent
In secondary preservation liquid, it is separately added into the E.C. 3.4.21.64 of different quality, forms E.C. 3.4.21.64 and the first mass volume ratio concentration preserving liquid
Different multiple finished products preserve liquid, are placed in the acceptance of the bid of differential responses container and remember clear, separately sampled from same sample put into each
Reaction vessel in, after reacting under the same conditions, record reaction result such as table 2；
Table 2:
| The mass volume ratio concentration of E.C. 3.4.21.64 | Process the effect of mucus | Impact (Quality Control IC point) on HPV result |
| 1mg/mL | Not exclusively | Part suppression |
| 2mg/mL | Not exclusively | Part suppression |
| 3mg/mL | Completely | Part suppression |
| 4mg/mL | Completely | Unrestraint phenomenon |
| 5mg/mL | Completely | Unrestraint phenomenon |
| 6mg/mL | Completely | Unrestraint phenomenon |
| 7mg/mL | Completely | Part suppression |
| 8mg/mL | Completely | Part suppression |
| 9mg/mL | Completely | Part suppression |
Reaction result is as shown in table 2: when E.C. 3.4.21.64 and the first mass volume ratio concentration preserving liquid be respectively 4mg/ml,
When 5mg/ml, 6mg/ml, mucus the testing result unrestraint phenomenon to HPV can be processed completely；
When E.C. 3.4.21.64 is respectively 3mg/ml, 7mg/ml, 8mg/ml, 9mg/ with the first mass volume ratio concentration preserving liquid
Mucus can be processed completely during ml and the testing result to HPV has part suppression；
When E.C. 3.4.21.64 is respectively 1mg/ml, 2mg/ml with the first mass volume ratio concentration preserving liquid, process mucus also
Not exclusively and part there is is to suppress the testing result of HPV；
In sum, preferred E.C. 3.4.21.64 is 4-6mg/ml with the first mass volume ratio concentration preserving liquid, most preferably
E.C. 3.4.21.64 and the first mass body volume concentrations preserving liquid ratio is for 5mg/ml.
Experimental example 4
Test the cervix cell preservation solution (Dodecyl trimethyl ammonium chloride quality in first preservation liquid of the present invention
Percentage ratio is 10%, and E.C. 3.4.21.64 is 5mg/ml with the first mass volume ratio concentration preserving liquid) contrast in the case of not eluting
(cell-preservation liquid that Chaozhou Kaipu Biochemistry Co., Ltd. produces, mainly becomes producer's cervix cell preservation solution of prior art
Point: PBS) in the case of eluting, in fluorescent quantitative PCR technique confirmatory experiment, whether produced curve and CT value have difference；
Before experiment, HPV.16 need to be carried out+PCR expands, and obtains standard curve, as shown in Figure 1.
During experiment, with 5 example cervix cells courage and uprightness samples, it is respectively adopted the present invention not eluting, producer's cell-preservation liquid eluting
Operation, uses the present invention not eluting and producer two kinds of operations of cell-preservation liquid eluting to be produced by fluorescent quantitative PCR technique checking
Raw curve and CT value have indifference, and as shown in figures 2-6, wherein: abscissa is period, vertical coordinate is copy to experimental result
Number；
Test result indicate that, curve produced by shown five example experiments is substantially overlapping with prior art, and this is described
Bright effectively and will not suppression PCR process.
Experimental example 5
(Dodecyl trimethyl ammonium chloride is 10% in the first mass percent preserved in liquid, egg to have tested the present invention
White enzyme K is 5mg/ml with the first mass volume ratio concentration preserving liquid) in the case of not eluting, contrast the producer of prior art
Cervix cell preservation solution (cell-preservation liquid that Chaozhou Kaipu Biochemistry Co., Ltd. produces, main component: PBS) at eluting and
Do not carry out in the case of eluting flow hybridization method checking type qualification test to verify the effect that the present invention is played,
As it is shown in fig. 7, this experimental result is represented by HPV probe thin film typing schematic diagram, typing schematic diagram is arranged by four rows six
Form composition, the upper left corner of form be for detection membrane bar whether the most effectively Biotin point, immediately below Biotin point
Whether grid is for existing the Quality Control IC point of mortifier, amplification efficiency and DNA concentration during detection PCR, remaining each grid is each
Represent a kind of HPV virus subtype.
Experimental result will be recorded on probe film bar, and probe film bar is provided with the form corresponding with typing schematic diagram,
Biotin point 1 is corresponding with the table position of HPV thin film typing schematic diagram respectively with IC point 2, the HPV virus subtype institute that inspection is arrived
The grid at place is also corresponding with HPV probe thin film typing schematic diagram.
Probe film bar after experiment as it is shown in fig. 7, a row has five groups of experiment sample 1001-1005, its from left to right,
One is sample 1001-1004 to the 4th, is containing blood sample, and the 5th is sample 1005, is free from blood sample, and three rows in Fig. 7 divide
The most corresponding three kinds of experimental conditions of type schematic diagram, first row is the experimental conditions of the cell-preservation liquid group not eluting of the present invention, the
Two rows are the experimental conditions after the cell-preservation liquid group eluting of prior art, and the cell-preservation liquid group of the 3rd row's prior art is not washed
De-experimental conditions, experimental result be described as follows shown in table 3:
Test result indicate that: after the experimental conditions of the cell-preservation liquid of the present invention and the cell-preservation liquid eluting of prior art
Experimental conditions consistent, and the cell-preservation liquid group of prior art HPV testing result in the case of not eluting can be suppressed.Real
Test result to indicate the cell-preservation liquid of the present invention and also be able to have with of the prior art under conditions of removing elution step from
Cell-preservation liquid HPV accuracy of detection after eluting processes.
Summing up above-mentioned experimental example, this cervix cell preservation solution has easy and simple to handle, and preparation is easy, to courageous and upright sample and mucus
The treatment effect of sample is preferably and advantage that suppression will not occur.
Claims (4)
1. a cervix cell preservation solution, it is characterised in that: include PBS, Dodecyl trimethyl ammonium chloride and albumen
Enzyme K, described Dodecyl trimethyl ammonium chloride is first mixed in PBS to be formed and preserves liquid for the first time, described dodecyl front three
Ammonium chloride is 5-15% at the first mass percent concentration preserved in liquid；Described first preservation it is mixed into after described E.C. 3.4.21.64
Liquid forms finished product and preserves liquid, and described E.C. 3.4.21.64 is 4-6mg/ml with the mass volume ratio concentration of described first preservation liquid.
Cervix cell preservation solution the most according to claim 1, it is characterised in that: described PBS is for tell containing 0.05%
The phosphate buffer of the pH=7.4 of temperature-20.
Cervix cell preservation solution the most according to claim 1, it is characterised in that: described Dodecyl trimethyl ammonium chloride exists
The first mass percent concentration preserved in liquid is 10%.
4. according to the cervix cell preservation solution described in claims 1 to 3 any claim, it is characterised in that: described E.C. 3.4.21.64
It is 5mg/ml with the mass volume ratio concentration of described first preservation liquid.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018113796A1 (en) * | 2016-12-23 | 2018-06-28 | 西比曼生物科技（上海）有限公司 | Cell freezing medium for clinical use |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101363011A (en) * | 2008-09-17 | 2009-02-11 | 上海艾迪康临床检验中心有限公司 | Cervical exfoliated cell preservative fluid |
| CN101485303A (en) * | 2009-02-26 | 2009-07-22 | 广州鸿琪光学仪器科技有限公司 | Exfoliated cell preservative fluid |
| CN103120153A (en) * | 2012-11-26 | 2013-05-29 | 刘召宏 | Exfoliative cells preserving fluid |
| CN105092328A (en) * | 2015-07-30 | 2015-11-25 | 厦门宝太生物科技有限公司 | Erythrocyte removing treating fluid and application |
-
2016
- 2016-09-30 CN CN201610875401.0A patent/CN106234354B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101363011A (en) * | 2008-09-17 | 2009-02-11 | 上海艾迪康临床检验中心有限公司 | Cervical exfoliated cell preservative fluid |
| CN101485303A (en) * | 2009-02-26 | 2009-07-22 | 广州鸿琪光学仪器科技有限公司 | Exfoliated cell preservative fluid |
| CN103120153A (en) * | 2012-11-26 | 2013-05-29 | 刘召宏 | Exfoliative cells preserving fluid |
| CN105092328A (en) * | 2015-07-30 | 2015-11-25 | 厦门宝太生物科技有限公司 | Erythrocyte removing treating fluid and application |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018113796A1 (en) * | 2016-12-23 | 2018-06-28 | 西比曼生物科技（上海）有限公司 | Cell freezing medium for clinical use |
| CN108235981A (en) * | 2016-12-23 | 2018-07-03 | 西比曼生物科技（上海）有限公司 | It is a kind of can Clinical practice cells frozen storing liquid |
| CN108235981B (en) * | 2016-12-23 | 2021-07-23 | 西比曼生物科技（香港）有限公司 | Cell cryopreservation liquid capable of being used clinically |
| US11363812B2 (en) | 2016-12-23 | 2022-06-21 | Cellular Biomedicine Group Hk Limited | Cell freezing medium for clinical use |
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| CN106234354B (en) | 2019-05-21 |
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| CB03 | Change of inventor or designer information |
Inventor after: Feng Jing Inventor after: Wei Quhao Inventor after: Xiao Linlin Inventor before: Xiao Linlin Inventor before: Feng Jing Inventor before: Wei Quhao |
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Effective date of registration: 20190417 Address after: No. 6600 Nanfeng Highway, Nanqiao Town, Fengxian District, Shanghai 201400 Applicant after: Shanghai Fengxian Centrl Hospital Applicant after: Xiao Linlin Address before: No. 6600 Nanfeng Highway, Nanqiao Town, Fengxian District, Shanghai 201400 Applicant before: Shanghai Fengxian Centrl Hospital |
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Effective date of registration: 20191128 Address after: 201400, Shanghai, Fengxian District South Village Road, No. 6600 Patentee after: Shanghai Fengxian Centrl Hospital Address before: 201400, Shanghai, Fengxian District South Village Road, No. 6600 Co-patentee before: Xiao Linlin Patentee before: Shanghai Fengxian Centrl Hospital |
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