CN202189009U - High-sensitivity chromogenic TAL kit - Google Patents
High-sensitivity chromogenic TAL kit Download PDFInfo
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- CN202189009U CN202189009U CN2011202152956U CN201120215295U CN202189009U CN 202189009 U CN202189009 U CN 202189009U CN 2011202152956 U CN2011202152956 U CN 2011202152956U CN 201120215295 U CN201120215295 U CN 201120215295U CN 202189009 U CN202189009 U CN 202189009U
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Abstract
The utility model discloses a high-sensitivity chromogenic TAL (Tachypleus Amebocyte Lysate) kit. The high-sensitivity chromogenic TAL kit comprises a kit body, as well as two bottles of TAL, two bottles of bacterial endotoxin working standard substance, two bottles of chromogenic substrate, two bottles of first diazotization reagent, two bottles of second diazotization reagent, two bottles of third diazotization reagent, a bottle of reaction terminator and two bottles of bacterial endotoxin test water in the kit body. The high-sensitivity chromogenic TAL kit can test the bacterial endotoxin with the sensitivity up to 0.01EU/ml, which is ten times higher than that of ordinary chromogenic TAL. Together with the diazotization coloring agent, the color generated by the chromogenic TAL changes from yellow to rose-red, so that interference on the chromogenic reaction from the colors of colored samples (such as jaundice and the like) is avoided, and the high-sensitivity chromogenic TAL kit has the advantages of high sensitivity, less interference and the like.
Description
Technical field
The utility model discloses a kind of high sensitivity colour developing tachypleus amebocyte lysate box, divides by International Patent Classification (IPC) (IPC) to belong to detection kit class manufacturing technology field, especially relates to a kind of endotoxin and detects tachypleus amebocyte lysate box.
Background technology
Bacterial endotoxin is claimed pyrogen again, can cause body generation pyrogen reaction behind the entering human body, causes endotoxemia, endotoxin shock, threat to life when serious.Limulus test is the endotoxic best approach of bacterial detection.Its principle is to contain the protease system responsive to bacterial endotoxin in the TAL, and this enzyme system comprises the C factor, the B factor, proclotting enzyme and coagulagen etc.Under the effect of bacterial endotoxin, inducing B factor activation after the activation of the C factor, causing the proclotting enzyme activation after the activation of the B factor is coagulase, and coagulase to cause the coagulagen activation be coagulated protein, coagulated protein aggregates into gel at last.For accurately quantitative bacterial endotoxin, can in reaction system, add the colour developing matrix of synthetic.This colour developing matrix is the substrate of coagulase, under the effect of coagulase, can generate yellow paranitroanilinum.In the regular hour, the growing amount of paranitroanilinum becomes positive correlation with bacterial endotoxin concentration, and the growing amount of paranitroanilinum can utilize optical instrument to detect at OD405nm.Therefore, can be accurately the quantitative endotoxin concns in the sample.The method is colour developing matrix method.The sensitivity of colour developing matrix method is generally 0.1EU/ml.When application development process TAL carries out the endotoxin detection of biological sample, usually can receive of the interference of coloured sample self color to LTOY LALT.As the jaundice sample that often has clinically also is yellow, and background OD value height can't detect with the colour developing TAL.
Summary of the invention
Deficiency to prior art; The utility model provides a kind of high sensitivity colour developing tachypleus amebocyte lysate box; Be to carry out the azo reaction through the product paranitroanilinum that after the reaction end of TAL and bacterial endotoxin, will develop the color to realize that the endotoxic sensitivity of this kit bacterial detection reaches 0.01EU/ml.
For achieving the above object, the utility model is realized through following technical scheme:
A kind of high sensitivity colour developing tachypleus amebocyte lysate box; Comprise that box body and interior TAL bottle thereof, bacterial endotoxin working standard bottle, colour developing matrix bottle, the azo reagent bottle more than two, reaction terminating agent bottle and bacterial endotoxin inspection use water bottle; The color of chromogenic reaction is a rose, and the highest absorption peak is 545nm.
Further, two bottles of TALs are arranged in the described box body, two bottles of bacterial endotoxin working standards, two bottles of colour developing matrix, three groups of azo reagent bottles, one bottle of reaction terminating agent and two bottles of bacterial endotoxin inspection waters.
Further, described three groups of azo reagent bottles are respectively sodium nitrite bottle, Amcide Ammate bottle and naphthodiamide bottle and quantity is two bottles.
Further, the colour developing matrix in the said colour developing matrix bottle is tripeptides or the tetrapeptide that has the p-nitrophenyl amine groups, and loading amount is 0.5ml-2.2ml.
Further, in the described reaction terminating agent bottle 0.48N hydrochloric acid is housed, loading amount is 50ml.
Further, described TAL bottle, bacterial endotoxin working standard bottle, colour developing matrix bottle, azo reagent bottle, reaction terminating agent bottle and bacterial endotoxin inspection are glass glass tube vial or ampoule bottle with water bottle.
The utility model is to carry out the azo reaction through the product paranitroanilinum that after the reaction end of TAL and bacterial endotoxin, will develop the color to realize; The color of chromogenic reaction is a rose; Sensitivity reaches 0.01EU/ml, and the bacterial endotoxin sizing technique that is specially adapted to clinical sample, biological sample and coloured sample detects.
The utlity model has following beneficial effect:
1, the endotoxic sensitivity of this kit bacterial detection reaches 0.01EU/ml, improves 10 times than common development process TAL;
2, the color of the utility model chromogenic reaction is a rose, and the highest absorption peak is 545nm, rather than traditional 405nm, has avoided the interference of most of biological sample self color to the development process LTOY LALT.
Description of drawings
Fig. 1 is the utility model structural representation.
Embodiment
Below in conjunction with accompanying drawing the utility model is described further:
Embodiment: see also Fig. 1, a kind of high sensitivity colour developing tachypleus amebocyte lysate box comprises two bottles of TALs 2 in its box body 1; 9, two bottle of first azo reagent 4 of two bottles of bacterial endotoxin working standard 3, two bottles of colour developings matrix; 6, one bottles of reaction terminating agents 7 of 5, two bottles of trisazo-reagent of two bottle of second azo reagent; Two bottles of bacterial endotoxin inspection waters 8, wherein
The TAL loading amount is 0.5ml-2.2ml, after freeze drying, is sealed in glass glass tube vial or the ampoule;
The bacterial endotoxin working standard, tiring is 5EU-200EU, after freeze drying, is sealed in glass glass tube vial or the ampoule;
Colour developing matrix is tripeptides or the tetrapeptide that has the p-nitrophenyl amine groups, and loading amount is 0.5ml-2.2ml, after freeze drying, is sealed in glass glass tube vial or the ampoule;
The first azo reagent is 0.04% sodium nitrite, after freeze drying, is sealed in glass glass tube vial or the ampoule;
The second azo reagent is 0.3% Amcide Ammate, after freeze drying, is sealed in glass glass tube vial or the ampoule;
Trisazo-reagent is 0.07% naphthodiamide, after freeze drying, is sealed in glass glass tube vial or the ampoule;
Reaction terminating agent is a 0.48N hydrochloric acid, and loading amount is 50ml, is sealed in the vial;
Bacterial endotoxin inspection water, loading amount is 50ml, is sealed in the vial.
The method of application of the utility model:
1. get one bottle of bacterial endotoxin working standard, add bacterial endotoxin inspection water, be diluted to 0.1EU/ml after the dissolving, 0.05EU/ml, 0.025EU/ml, 0.01EU/ml endotoxin standard solution;
2. press the kit labelled amount with bacterial endotoxin inspection water dissolving TAL, colour developing matrix, second azo reagent and the trisazo-reagent;
3. press the kit labelled amount with the reaction terminating agent dissolving with hydrochloric acid first azo reagent;
4. get the apyrogeneity test tube, each pipe adds 100 μ l negative controls (being bacterial endotoxin inspection water), endotoxin standard solution respectively, or sample;
5. each pipe adds 100 μ l TAL solution, mixing, 37oC incubation 30-60 minute respectively;
6. each pipe adds 100 μ l colour developing matrix solution, mixing, 37oC incubation 3-8 minute respectively;
7. incubation finishes, and each pipe adds 500 μ l, the first azo reagent solution, mixing respectively;
8. each pipe adds 500 μ l, the second azo reagent solution, mixing respectively;
9. each pipe adds 500 μ l trisazo-reagent solutions respectively, and mixing left standstill 5 minutes, utilized optical detecting instrument (like spectrophotometer) to read the OD value in the 545nm wavelength;
10. set up typical curve according to the OD value of endotoxin standard solution: Y=b X+a, wherein Y is the OD of a 545nm place value, and X is the concentration of endotoxin solution, and b is a straight slope, and a is the Y y-intercept;
11., on typical curve, draw the endotoxin concns of sample per sample at the OD at 545nm place value reading.
The supporting azo coloring agent of the utility model; The color that the colour developing TAL is produced is changed into rose by yellow; Can avoid the interference of the color of coloured sample (like jaundice etc.) to chromogenic reaction, the highest absorption peak is 545nm, and the endotoxic sensitivity of bacterial detection reaches 0.01EU/ml.
The above record is merely the embodiment that utilizes this origination techniques content, anyly is familiar with modification, the variation that this creation of this art utilization is done, and all belongs to the claim of this creation opinion, and is not limited to embodiment announcement person.
Claims (5)
1. high sensitivity colour developing tachypleus amebocyte lysate box; It is characterized in that: comprise that box body and interior TAL bottle thereof, bacterial endotoxin working standard bottle, colour developing matrix bottle, the azo reagent bottle more than two, reaction terminating agent bottle and bacterial endotoxin inspection use water bottle; The color of chromogenic reaction is a rose, and the highest absorption peak is 545nm.
2. high sensitivity colour developing tachypleus amebocyte lysate box according to claim 1; It is characterized in that: two bottles of TALs are arranged, two bottles of bacterial endotoxin working standards, two bottles of colour developing matrix in the described box body; Three groups of azo reagent bottles, one bottle of reaction terminating agent and two bottles of bacterial endotoxin inspection waters.
3. high sensitivity colour developing tachypleus amebocyte lysate box according to claim 2, it is characterized in that: described three groups of azo reagent bottles are respectively sodium nitrite bottle, Amcide Ammate bottle and naphthodiamide bottle and quantity is two bottles.
4. high sensitivity colour developing tachypleus amebocyte lysate box according to claim 1 and 2, it is characterized in that: the colour developing matrix in the said colour developing matrix bottle is tripeptides or the tetrapeptide that has the p-nitrophenyl amine groups, and loading amount is 0.5ml-2.2ml.
5. high sensitivity colour developing tachypleus amebocyte lysate box according to claim 1 and 2, it is characterized in that: in the described reaction terminating agent bottle 0.48N hydrochloric acid is housed, loading amount is 50ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202152956U CN202189009U (en) | 2011-06-23 | 2011-06-23 | High-sensitivity chromogenic TAL kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202152956U CN202189009U (en) | 2011-06-23 | 2011-06-23 | High-sensitivity chromogenic TAL kit |
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| Publication Number | Publication Date |
|---|---|
| CN202189009U true CN202189009U (en) | 2012-04-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011202152956U Expired - Lifetime CN202189009U (en) | 2011-06-23 | 2011-06-23 | High-sensitivity chromogenic TAL kit |
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| CN (1) | CN202189009U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290890A (en) * | 2016-07-15 | 2017-01-04 | 科赫生物科技（北京）有限公司 | The human body fluid development process fungus 1,3 β D glucosan detection kit of a kind of improvement and application process thereof |
| WO2019138211A1 (en) * | 2018-01-09 | 2019-07-18 | Molendotech Limited | Water quality testing |
-
2011
- 2011-06-23 CN CN2011202152956U patent/CN202189009U/en not_active Expired - Lifetime
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290890A (en) * | 2016-07-15 | 2017-01-04 | 科赫生物科技（北京）有限公司 | The human body fluid development process fungus 1,3 β D glucosan detection kit of a kind of improvement and application process thereof |
| CN106290890B (en) * | 2016-07-15 | 2018-10-23 | 科赫生物科技（北京）有限公司 | A kind of the human body fluid development process fungi 1,3- calloses detection kit and its application process of improvement |
| WO2019138211A1 (en) * | 2018-01-09 | 2019-07-18 | Molendotech Limited | Water quality testing |
| GB2571613A (en) * | 2018-01-09 | 2019-09-04 | Molendotech Ltd | Water Quality testing |
| CN111771118A (en) * | 2018-01-09 | 2020-10-13 | 莫兰多科技有限公司 | Water quality testing |
| GB2571613B (en) * | 2018-01-09 | 2021-01-27 | Molendotech Ltd | Water Quality testing |
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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 Jia new Patentee after: Xiamen limulus reagent biological Polytron Technologies Inc Address before: 361000 Fujian Province, Xiamen City Haicang Xinyang Industrial Zone No. 131 Jia new Patentee before: Company limited of tachypleus amebocyte lysate trial (demonstration) plant of Xiamen City |
|
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20120411 |