CN203965445U - A kind of dynamic color method limulus kit for endotoxin detection - Google Patents
A kind of dynamic color method limulus kit for endotoxin detection Download PDFInfo
- Publication number
- CN203965445U CN203965445U CN201420377086.5U CN201420377086U CN203965445U CN 203965445 U CN203965445 U CN 203965445U CN 201420377086 U CN201420377086 U CN 201420377086U CN 203965445 U CN203965445 U CN 203965445U
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- CN
- China
- Prior art keywords
- amebocyte lysate
- tachypleus amebocyte
- bottles
- endotoxin
- bottle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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- 239000002158 endotoxin Substances 0.000 title claims abstract description 31
- 241000239218 Limulus Species 0.000 title claims abstract description 21
- 238000001514 detection method Methods 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 18
- 241000239222 Tachypleus Species 0.000 claims abstract description 36
- 239000006166 lysate Substances 0.000 claims abstract description 36
- 239000003593 chromogenic compound Substances 0.000 claims abstract description 10
- 210000000601 blood cell Anatomy 0.000 claims abstract description 4
- 239000013592 cell lysate Substances 0.000 claims abstract description 4
- 239000011812 mixed powder Substances 0.000 claims abstract description 4
- TYMLOMAKGOJONV-UHFFFAOYSA-N 4-nitroaniline Chemical group NC1=CC=C([N+]([O-])=O)C=C1 TYMLOMAKGOJONV-UHFFFAOYSA-N 0.000 claims description 3
- 210000002381 plasma Anatomy 0.000 abstract description 3
- 206010003445 Ascites Diseases 0.000 abstract description 2
- 210000001124 body fluid Anatomy 0.000 abstract description 2
- 239000010839 body fluid Substances 0.000 abstract description 2
- 210000001175 cerebrospinal fluid Anatomy 0.000 abstract description 2
- 210000000038 chest Anatomy 0.000 abstract description 2
- 238000004108 freeze drying Methods 0.000 abstract description 2
- 210000002700 urine Anatomy 0.000 abstract description 2
- 238000005259 measurement Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 32
- 239000000243 solution Substances 0.000 description 16
- 230000004913 activation Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 239000000376 reactant Substances 0.000 description 3
- 108010065152 Coagulase Proteins 0.000 description 2
- 108090000056 Complement factor B Proteins 0.000 description 2
- 102000003712 Complement factor B Human genes 0.000 description 2
- 239000012190 activator Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 108010048121 pro-clotting enzyme Proteins 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 231100000284 endotoxic Toxicity 0.000 description 1
- 230000002346 endotoxic effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model discloses a kind of dynamic color method limulus kit for endotoxin detection, comprise box body and interior tachypleus amebocyte lysate bottle and sample treatment solution bottle thereof; The freeze-dried mixed powder of tachypleus amebocyte lysate and chromogenic substrate is housed in described tachypleus amebocyte lysate bottle; Described tachypleus amebocyte lysate is the tachypleus amebocyte lysate to bacterial endotoxin idiosyncrasy of extracting in limulus blood cell lysate.The utility model is by chromogenic substrate and tachypleus amebocyte lysate freeze-drying in same container, and application dynamic color method realizes the quantitative measurement of endotoxin of the body fluid such as clinical blood plasma, urine, cerebrospinal fluid, chest ascites is detected, a step, and highly sensitive, antijamming capability is strong.
Description
Technical field
The utility model belongs to the technical field that detection kit class is manufactured, and relates in particular to a kind of dynamic color method limulus kit for endotoxin detection.
Background technology
The C factor to bacterial endotoxin sensitivity, Factor B, proclotting enzyme, the coagulagen isoreactivity enzyme that in tachypleus amebocyte lysate, contain.Under applicable condition, the C factor is activated by bacterial endotoxin, cause C factor bypass reaction, both the C factor generated the activation C factor, activation C factor activator Factor B generates the activated b factor, activated b factor activator proclotting enzyme generates the coagulase of activation, and the coagulase of activation activates coagulagen and generates coagulated protein, and final set albumen aggregates into gel.The limulus test of setting up according to this principle is the endotoxic best approach of bacterial detection.According to reactant liquor turbidity, rising carrys out quantitative bacterial endotoxin at present conventional dynamic turbidimetric tachypleus amebocyte lysate.Owing to containing the interference factor that disturbs coagulated protein to form gel in common clinical sample, therefore dynamic turbidimetric sensitivity is not high, is easily disturbed.
In view of this, the inventor studies and has designed a kind of dynamic color method limulus kit for endotoxin detection, and this case produces thus.
Utility model content
The purpose of this utility model is to provide a kind of dynamic color method limulus kit for endotoxin detection, adopts chromogenic reaction quantitatively to detect clinical sample, aggregates into gel because chromogenic reaction does not rely on coagulated protein, and highly sensitive, antijamming capability is strong; Adopt sample treatment solution to dilute heat treated to clinical sample to be tested, further to get rid of interference simultaneously.
In order to solve above-mentioned purpose, the technical scheme that the utility model adopts is:
A kind of dynamic color method limulus kit for endotoxin detection, comprises box body and interior tachypleus amebocyte lysate bottle and sample treatment solution bottle thereof; The freeze-dried mixed powder of tachypleus amebocyte lysate and chromogenic substrate is housed in described tachypleus amebocyte lysate bottle.
Further, described tachypleus amebocyte lysate is the tachypleus amebocyte lysate to bacterial endotoxin idiosyncrasy of extracting in limulus blood cell lysate.
Further, described chromogenic substrate is tripeptides or the tetrapeptide with p-nitrophenyl amine groups.
Further, in described box body, have some bottles of tachypleus amebocyte lysate bottles and some bottles of sample treatment solution bottles, and the quantitative proportion of described tachypleus amebocyte lysate bottle and described sample treatment solution bottle is 1:1.
Further, in described box body, there are 20 bottles of tachypleus amebocyte lysate bottles and 20 bottles of sample treatment solution bottles.
The utlity model has following beneficial effect:
1, by freeze-dried mixed in a container to chromogenic substrate and tachypleus amebocyte lysate, do not need other supporting chromogenic substrate when use, single step reaction gets final product accurate quantitative analysis endotoxin, is applicable to mass, the detection of robotization endotoxin.
2, adopting dynamic color method to realize bacterial endotoxin quantitatively detects, the method does not need to rely on coagulated protein to aggregate into quantitatively bacterial endotoxin of gel, and therefore, antijamming capability is strong, the endotoxin that is applicable to the biological specimens such as clinical plasma sample detects, and detection sensitivity is up to 0.001EU/ml.
3, supporting sample treatment solution, carries out ten times of dilutions heat treated to clinical detection sample, further eliminates the interference of clinical sample to be tested to limulus test in dilution.
Brief description of the drawings
Fig. 1 is structural representation of the present utility model.
Embodiment
As shown in Figure 1, the utility model has disclosed a kind of dynamic color method limulus kit for endotoxin detection, comprises box body 1 and interior tachypleus amebocyte lysate bottle 11 and sample treatment solution bottle 12 thereof; The freeze-dried mixed powder of tachypleus amebocyte lysate and chromogenic substrate is housed in described tachypleus amebocyte lysate bottle 11.
Further, described tachypleus amebocyte lysate is the tachypleus amebocyte lysate to bacterial endotoxin idiosyncrasy of extracting in limulus blood cell lysate.
Further, described chromogenic substrate is tripeptides or the tetrapeptide with p-nitrophenyl amine groups.
Further, in described box body 1, have some bottles of tachypleus amebocyte lysate bottles 11 and some bottles of sample treatment solution bottles 12, and the quantitative proportion of described tachypleus amebocyte lysate bottle 11 and described sample treatment solution bottle 12 is 1:1.
Further, described dynamic color method limulus kit for endotoxin detection, has 11 and 20 bottles of sample treatment solution bottles 12 of 20 bottles of tachypleus amebocyte lysate bottles in box body 1.
Described sample treatment solution is used for diluting clinical sample to be tested, and the volume ratio of sample treatment solution and sample to be tested is 9:1, is diluted ten times in the time that clinical sample to be tested adds sample treatment solution.And in sample treatment solution, contain and eliminate the special composition that clinical sample disturbs, in diluted sample, further eliminate the interference of clinical sample to be tested to limulus test.
Using method of the present utility model:
1. clinical sample pre-service: get clinical sample to be tested 0.1ml and add in sample treatment solution bottle (containing sample treatment solution 0.9ml), mix, be placed in 65-110 ° of C heating 3-10 minute.Heat complete, immediately take out be placed in ice-water bath cooling 3 minutes.
2. machine on application of sample: get step 1 gained sample 0.2ml and join tachypleus amebocyte lysate bottle (loading amount is 0.2ml, is sealed in glass glass tube vial or ampoule after freeze drying), jog dissolves tachypleus amebocyte lysate.Immediately reactant liquor is placed in to the optically-read instrument with incubating device, 37 ° of C incubation reaction, and the dynamic change of recording the OD value of 405nm with dynamics software.
3. rise to default OD required time according to reactant liquor OD value and automatically draw endotoxin content by software.
The utility model is easy to use, after being diluted to heat treated, mixes with tachypleus amebocyte lysate clinical sample, being placed in just can automatic ration bacterial endotoxin with the optically-read instrument of incubating device, being applicable to mass robotization detects, in addition, the utility model does not need to rely on the formation of gel to carry out quantitative bacterial endotoxin, and detection sensitivity is up to 0.001EU/ml, antijamming capability is strong, and the endotoxin that is applicable to the body fluid biological specimens such as clinical blood plasma, urine, cerebrospinal fluid, chest ascites detects.
The above, it is only the utility model preferred embodiment, therefore can not limit successively the scope that the utility model is implemented, the equivalence of doing according to the utility model the scope of the claims and description changes and modifies, and all should still belong in the scope that the utility model contains.
Claims (5)
1. a dynamic color method limulus kit for endotoxin detection, is characterized in that: comprise box body and interior tachypleus amebocyte lysate bottle and sample treatment solution bottle thereof; The freeze-dried mixed powder of tachypleus amebocyte lysate and chromogenic substrate is housed in described tachypleus amebocyte lysate bottle.
2. a kind of dynamic color method limulus kit for endotoxin detection as claimed in claim 1, is characterized in that: described tachypleus amebocyte lysate is the tachypleus amebocyte lysate to bacterial endotoxin idiosyncrasy of extracting in limulus blood cell lysate.
3. a kind of dynamic color method limulus kit for endotoxin detection as claimed in claim 1, is characterized in that: described chromogenic substrate is with the tripeptides of p-nitrophenyl amine groups or tetrapeptide.
4. a kind of dynamic color method limulus kit for endotoxin detection as claimed in claim 1, it is characterized in that: in described box body, have some bottles of tachypleus amebocyte lysate bottles and some bottles of sample treatment solution bottles, and the quantitative proportion of described tachypleus amebocyte lysate bottle and described sample treatment solution bottle is 1:1.
5. a kind of dynamic color method limulus kit for endotoxin detection as claimed in claim 1, is characterized in that: in described box body, have 20 bottles of tachypleus amebocyte lysate bottles and 20 bottles of sample treatment solution bottles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420377086.5U CN203965445U (en) | 2014-07-09 | 2014-07-09 | A kind of dynamic color method limulus kit for endotoxin detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420377086.5U CN203965445U (en) | 2014-07-09 | 2014-07-09 | A kind of dynamic color method limulus kit for endotoxin detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN203965445U true CN203965445U (en) | 2014-11-26 |
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| CN201420377086.5U Expired - Lifetime CN203965445U (en) | 2014-07-09 | 2014-07-09 | A kind of dynamic color method limulus kit for endotoxin detection |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105445467A (en) * | 2015-11-27 | 2016-03-30 | 中国大冢制药有限公司 | Method for detecting bacterial endotoxin of sodium metabisulfite |
| CN105510590A (en) * | 2015-11-27 | 2016-04-20 | 中国大冢制药有限公司 | Detection method for cysteine hydrochloride bacterial endotoxin |
| CN113302494A (en) * | 2018-10-21 | 2021-08-24 | 厦门鲎试剂生物科技股份有限公司 | Novel method for rapidly detecting sepsis by using gram-negative bacterial infection |
-
2014
- 2014-07-09 CN CN201420377086.5U patent/CN203965445U/en not_active Expired - Lifetime
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105445467A (en) * | 2015-11-27 | 2016-03-30 | 中国大冢制药有限公司 | Method for detecting bacterial endotoxin of sodium metabisulfite |
| CN105510590A (en) * | 2015-11-27 | 2016-04-20 | 中国大冢制药有限公司 | Detection method for cysteine hydrochloride bacterial endotoxin |
| CN105445467B (en) * | 2015-11-27 | 2017-07-28 | 中国大冢制药有限公司 | The detection method of sodium pyrosulfite bacterial endotoxin |
| CN113302494A (en) * | 2018-10-21 | 2021-08-24 | 厦门鲎试剂生物科技股份有限公司 | Novel method for rapidly detecting sepsis by using gram-negative bacterial infection |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 361000 Fujian Province, Xiamen City Haicang Xinyang Industrial Zone No. 131 new Jia (plant) Patentee after: XIAMEN BIOENDO TECHNOLOGY Co.,Ltd. Address before: 361000 Fujian Province, Xiamen City Haicang Xinyang Industrial Zone No. 131 new Jia (plant) Patentee before: CHINESE HORSESHOE CRAB REAGENT MANUFACTORY Co.,Ltd. XIAMEN |
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| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20141126 |